Prospective observational study for perioperative volume replacement with 6% HES 130/0,42, 4% gelatin and 6% HES 200/0,5 in cardiac surgery.

BACKGROUND: the constantly growing amount of different kinds of colloid fluids necessitates comparative investigations with regards to the safety and effectivity in clinical use of these preparations. Hence we compared three colloid fluids in an observational study. The objective was the exploration of the influence of these three colloids on blood coagulation, hemodynamics and renal function of the cardiac surgical patient. METHODS: we included 90 patients undergoing an elective open-heart surgery with the use of the heart-lung machine and observed them consecutively. Group 1 [gelatin 4% (n = 30)], Group 2 [HES 200/0,5 (n = 30)] and Group 3 [HES 130/0,42 (n = 30)]. We measured the perioperative volume replacement, the administration of blood- and coagulation-products, the application of catecholamines, the renal function, blood gas and the platelet aggregation using multiplate electrode analyzer (Multiplate, Dynabyte medical, Munich, Germany). RESULTS: the gelatin-group needed significantly more norepinephrine than the HES 130/0.42 group. The responsible surgeon considered the blood coagulation in the HES 200/0.5 group most frequently as impaired. Furthermore we saw a significant decrease in platelet function in the HES 200/0.5 group when performing the multiplate-analysis (ADP-and COL-test). HES 130/0.4 as well as gelatin 4% showed no significant change in platelet function. The gelatin-group and the HES 200/0.5 needed significantly more aprotinine than the HES 130/0.4 group. We saw no significant difference with regards to administration of blood and coagulation products between the three groups. The urinary excretion during the intervention was significantly higher in the HES 200/0.5 group and in the gelatin group than in the HES 130/0.4 group. CONCLUSIONS: our results confirm the lower stabilizing effect of gelatin on circulation during fluid resuscitation. The blood coagulation was mostly impaired due to HES 200/0.5 confirmed by the multiplate®-analysis as well as by different clinical findings.

Functional consequences of reduction in NMDA receptor glycine affinity in mice carrying targeted point mutations in the glycine binding site.

We have used site-directed mutagenesis in conjunction with homologous recombination to generate two mouse lines carrying point mutations in the glycine binding site of the NMDAR1 subunit (Grin1). Glycine concentration-response curves from acutely dissociated hippocampal neurons revealed a 5- and 86-fold reduction in receptor glycine affinity in mice carrying Grin1(D481N) and Grin1(K483Q) mutations, respectively, whereas receptor glutamate affinity remained unaffected. Homozygous mutant Grin1(D481N) animals are viable and fertile and appear to develop normally. However, homozygous mutant Grin1(K483Q) animals are significantly lighter at birth, do not feed, and die within a few days. No gross abnormalities in CNS anatomy were detected in either Grin1(D481N) or Grin1(K483Q) mice. Interestingly, in situ hybridization and Western blot analysis revealed changes in the expression levels of NMDA receptor subunits in Grin1(D481N) mice relative to wild type that may represent a compensatory response to the reduction in receptor glycine affinity. Grin1(D481N) mice exhibited deficits in hippocampal theta burst-induced long-term potentiation (LTP) and spatial learning and also a reduction in sensitivity to NMDA-induced seizures relative to wild-type controls, consistent with a reduced activation of NMDA receptors. Mutant mice exhibited normal prepulse inhibition but showed increased startle reactivity. Preliminary analysis indicated that the mice exhibit a decreased natural aversion to an exposed environment. The lethal phenotype of Grin1(K483Q) animals confirms the critical role of NMDA receptor activation in neonatal survival. A milder reduction in receptor glycine affinity results in an impairment of LTP and spatial learning and alterations in anxiety-related behavior, providing further evidence for the role of NMDA receptor activation in these processes.

Anxiolytic-like activity of the mGLU2/3 receptor agonist LY354740 in the elevated plus maze test is disrupted in metabotropic glutamate receptor 2 and 3 knock-out mice.

RATIONALE: (1S,2S,5R,6S)-2-Aminobicyclo[3.1.0]hexane-2,6-dicarboxylic acid (LY354740) is a potent and selective agonist for group II metabotropic glutamate (mGlu2 and mGlu3) receptors, with anxiolytic-like activity in animal and human models, and efficacy in anxiety patients. However, the lack of mGlu2 or mGlu3 receptor specific agonists has prevented in vivo characterization of individual functions of these two receptors in mediating the anxiolytic-like effects of LY354740. OBJECTIVE: To utilize mGlu2 receptor and mGlu3 receptor knockout animals and the mGlu2/3 selective antagonist (2S,1'S,2'S)-2-(9-xanthylmethyl)-2-(2'-carboxycyclopropyl)glycine (LY341495) to further investigate the roles of mGlu2 and mGlu3 receptors in mediating the anxiolytic-like actions of LY354740 in a mouse model of anxiety [elevated plus maze (EPM) test]. METHODS: To confirm that mGlu2/3 receptors are responsible for anxiolytic-like activity in the EPM under these test conditions, mice were pretreated with LY341495 at 30 min prior to s.c. administered LY354740. Subsequently, saline vehicle or LY354740 was administered (s.c.) 30 min before the EPM testing in wild-type, mGlu2 receptor knockout, and mGlu3 receptor knockout mice. RESULTS: LY354740 reduced in a dose-dependent manner anxiety-related behavior on the EPM in wild-type mice with a maximally effective dose of 10--20 mg/kg s.c. Pretreatment with LY341495 potently prevented the anxiolytic-like effects of LY354740 (20 mg/kg, s.c.) in mice. Although the mGlu2 receptor knockout and mGlu3 receptor knockout mice were grossly normal, the anxiolytic-like activity of LY354740 (20 mg/kg, s.c.) was not evident in either mGlu2 or mGlu3 receptor knockout mice, when compared to their wild-type controls. CONCLUSIONS: The activation of both mGlu2 and mGlu3 receptors by LY354740 appears to be required for anxiolytic-like activity in the EPM test in mice. These studies serve as a foundation for additional studies on underlying circuits, brain structures, and receptor subtypes involved in the anxiolytic-like actions of mGlu receptor active agents, and the design of future drugs for anxiety disorders in humans.

In vitro chemoseparation of leukemic cells from murine bone marrow using VP16-213: importance of stem cell assays.

The use of chemopurified autologous bone marrow (BM) is being explored as a transplant source for patients with leukemia who do not have a HLA-matched donor. Because stem cell assays have not previously been found to predict engraftment after transplantation, the optimal drug(s) and drug concentrations have not been determined. To determine the effectiveness of the podophyllotoxin derivative, VP16, and the value of stem cell assays in chemopurification studies, a murine model using the C57B1/6 mouse and its syngeneic leukemia EL-4 was developed. Kill of committed (CFU-C) and pluripotent (CFU-S) hematopoietic stem cells and tumor (tCFU) stem cells after a 1-h exposure to VP16 was first determined. A marked kill differential of tCFU compared to that of the CFU-C/S populations was found, with no tCFU surviving at VP16 concentrations greater than 30 micrograms/ml. No kill differential of CFU-C versus CFU-S was seen at VP16 doses greater than 10 micrograms/ml. All mice transplanted with a mixture of 25 X 10(6) BM cells and 1 X 10(7) EL-4 cells treated in vitro with 40 micrograms/ml of VP16 died of their tumors. However by reducing the tumor burden to 1 X 10(6) EL-4 and 5 X 10(6) EL-4, nine of ten and four of six of the mice, respectively, survived 90+ days tumor free. On the basis of survival data, it was found that engraftment of VP16-treated BM was directly proportional to the product of the degree of CFU-S inhibition and BM cell inoculum, i.e., the number of viable CFU-S transplanted. The maximum VP16 concentration that led to predictable engraftment at BM doses less than 25 X 10(6) cells was 55 micrograms/ml. Thus, 15 mice were transplanted with 1 X 10(7) BM cells and 1 X 10(7) EL-4 cells, incubated with 55 micrograms/ml of VP16; 13 out of 15 survived tumor free for 90+ days. When VP16 was used as a BM chemopurification agent, up to 50% of contaminating tumor cells were eliminated from BM suspensions without affecting engraftment after transplantation. Because stem cell inhibition predicted engraftment, drug concentrations that maximized tumor cell kill could be chosen.

Profiles of adolescents' clothing practices: purchase, daily selection, and care.

The purpose of this study was to examine the clothing practices in the daily selection, care, and purchase of clothing by adolescents in order to determine the extent these practices are performed independently or influenced by others, and to identify the factors involved in the activities. Clothing Practice Profiles were developed for both sexes in the three age groups corresponding to divisions in the Oregon 4-H program. The age group comparison revealed increasing frequency of independent activity in all three clothing practice areas (selection, care, and purchase) as age increased. Parental influence appeared to decrease with increasing age. Siblings' influence was minimal. Peer influence on selection and purchase practices increased. Media influence on daily clothing selection practices and clothing purchase practices in terms of wearing or buying identical or similar clothing was minimal. Media influence in advertisements increased with age, but purchases of advertised clothing items remained about the same. Age group comparisons were also made for selection factors and other care and purchase practices, including responsibilities for care of the member's and family's clothing, planning clothing purchases, sources of funds and methods of paying for clothing, use of clothing label information, and purchase factors.

Renal transplant failure due to urologic complications: Comparison of static fluid with contrast-enhanced magnetic resonance urography.

PURPOSE: Postrenal reasons of renal transplant failure can be assessed by magnetic resonance urography. This study was designed to retrospectively compare the diagnostic accuracy of static fluid (T2-)MRU compared to contrast enhanced (CE-)MRU in patients with renal transplant failure. MATERIAL AND METHODS: Thirty-five consecutive patients (14 female, 21 men; mean age 48.6 years) with renal transplant failure and sonographically detected hydronephrosis were examined both with T2-MRU as well as CE-MRU resulting in 39 MRU examinations. MRU was performed both using T2-weighted HASTE-sequence (T2-MRU) as well as Gadolinium-enhanced 3D-FLASH-sequence (CE-MRU) on a 1.5-T clinical MRI scanner (Magnetom Vision, Siemens Medical Solutions). Subjective image quality of resulting maximum intensity projection was assessed in consensus by two readers blinded to the final diagnosis, using a five point scale. MRU findings were correlated to sonography, operative results or clinical follow up. RESULTS: CE-MRU yielded a sensitivity of 85.7% (T2-MRU 76.2%), and a specificity of 83.3% (T2-MRU: 73.7%), however statistical significance was not reached. The subjective image quality was significantly better in CE-MRU. CONCLUSIONS: Only concerning subjective image quality CE-MRU proved superior to T2-MRU. Yet, there was no significant difference in diagnostic accuracy between T2- and CE-MRU. Thinking of incipient nephrogenic systemic fibrosis, T2-MRU can be used as reliable alternative in patients with decreased renal transplant function due to urological complications.

Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing.

To develop a simplified method of bone marrow (BM) cryopreservation, changes were made in the standard method in three areas: the cryoprotectant, the method of cell freezing, and the storage temperature. Unfractionated BM cells from 60 patients were cryopreserved in 300-mL aliquots in both dimethylsulfoxide (DMSO) and hydroxyethyl starch (HES), a combination known to preserve granulocytes successfully. The cells were frozen without rate-controlled freezing by simple immersion into a -80 degrees C freezer where they remained until the time of reinfusion. The 60 patients underwent 72 autologous transplants after three high-dose chemotherapy regimens: 30 received high-dose carmustine in combination, five received high-dose busulfan and cyclophosphamide, and 37 received high-dose aziridinylbenzoquinone. The BM was infused for more than 30 minutes after rapid thawing at 37 degrees C. The mean post-thaw nucleated cell recovery was 96% +/- 11.6%, and Trypan blue dye exclusion was 82.2% +/- 9.2%. The mean postthaw CFU-GM and BFU-E recoveries were 81.9% +/- 39.0% and 90.5% +/- 41.2%. Complete count recovery occurred in 68 of 72 transplants. Median times to a WBC count greater than 1,000/microL, a granulocyte count greater than 1,000/microL and a platelet count greater than 20,000/microL were 15, 21, and 15 days, respectively. Risk factors for delayed recovery were not found. Unfractionated BM cells can be successfully cryopreserved in the DMSO/HES mixture rapidly and inexpensively, without rate-controlled freezing or storage at liquid nitrogen temperatures.

A radiolabeled antiglobulin test for crossmatching platelet transfusions.

Despite the use of HLA-matched platelets for alloimmunized recipients, transfusion failures occur. In order to reduce these failures, we investigated the use of a radiolabeled antiglobulin technique for platelet crossmatching. The principle of the test is that of an indirect Coombs test using 125I labeled goat anti-human IgG. Incompatibility is determined by calculating a radioactivity antiglobulin test (RAGT) index. Using this technique, we performed 89 crossmatches on 19 leukemic or aplastic patients who were refractory to random donor platelets and receiving varying degrees of HLA-matched platelets. Effectiveness of the transfusion was assessed from the posttransfusion corrected platelet count increment (CCI) determined at 1 and 20 hr. When the RAGT index was 1.9 or less, the mean CCI at 1 lhr was 17,570 +/- 7003/cu mm, n = 55. When the RAGT index was 2.0 or greater, the mean CCI was 4237 +/- 4100/cu mm, n = 34. At 20 hr when the RAGT index was 1.9 or less, the mean CCI was 8722 +/- 3143/cu mm, n = 33, and when the index was 2.0 or greater, the mean CCI was 571 +/- 1286/cu mm, n = 23. Using this technique, one false negative resulted. Nine positive crossmatches with good increments at 1 hr were found; at 20 hr, however, the survival of these units was zero. These data suggest that this method is a useful adjunct in the selection of platelets in the refractory patient.