Identification of microglial signal transduction pathways mediating a neurotoxic response to amyloidogenic fragments of beta-amyloid and prion proteins

Microglial interaction with amyloid fibrils in the brains of Alzheimer's and prion disease patients results in the inflammatory activation of these cells. We observed that primary microglial cultures and the THP-1 monocytic cell line are stimulated by fibrillar beta-amyloid and prion peptides to activate identical tyrosine kinase-dependent inflammatory signal transduction cascades. The tyrosine kinases Lyn and Syk are activated by the fibrillar peptides and initiate a signaling cascade resulting in a transient release of intracellular calcium that results in the activation of classical PKC and the recently described calcium-sensitive tyrosine kinase PYK2. Activation of the MAP kinases ERK1 and ERK2 follows as a subsequent downstream signaling event. We demonstrate that PYK2 is positioned downstream of Lyn, Syk, and PKC. PKC is a necessary intermediate required for ERK activation. Importantly, the signaling response elicited by beta-amyloid and prion fibrils leads to the production of neurotoxic products. We have demonstrated in a tissue culture model that conditioned media from beta-amyloid- and prion-stimulated microglia or from THP-1 monocytes are neurotoxic to mouse cortical neurons. This toxicity can be ameliorated by treating THP-1 cells with specific enzyme inhibitors that target various components of the signal transduction pathway linked to the inflammatory responses.     

Effects of fat supplementation and immature alfalfa to concentrate ratio on plasma progesterone, energy balance and reproductive traits of dairy cattle

Forty-six multiparous Holstein cows were assigned at 5 d postpartum to a completely randomized design employing a 2 x 3 factorial arrangement of treatments. Factors were 0 and 5% added prilled long-chain fatty acids (DM basis) and three forage to concentrate ratios (45:55, 64:36, 84:16). Diets consisted of immature alfalfa silage and a concentrate of shelled corn and soybean meal with or without fat replacing a portion of the corn. Mean plasma concentration of cholesterol was higher for cows fed 5% vs. 0% fat and increased over the first 100 d in milk for all animals regardless of treatment. There were no differences in reproductive performance due to either of the main effects. Mean plasma progesterone was higher due to fat treatment in the mid to late luteal phase of the second postpartum cycle as well as the metestrous to early luteal phase and mid to late luteal phase of the third cycle. Even though progesterone concentrations were higher in cows fed 5% fat during the luteal phase after breeding, the conception rates at this service were not different from those fed 0% fat. The biological significance of increased plasma progesterone concentration was not identified with any postpartum reproductive trait measured in this trial.     

Response to recombinant bovine somatotropin in dairy cows with different genetic merit for milk production

Thirty-nine multiparous cows obtained from two genetic lines were utilized to determine the effect of genetic merit on lactation response to long-term administration of recombinant bST. Cow index ranged from -70 to 456 (mean = 183) and -494 to -88 (mean = -288) kg milk for high and low genetic groups, respectively. Cows were blocked by calving date and randomly assigned to treatment within genetic group. Treatments were 0, 10.3, 20.6, and 30.9 mg somatotropin injected daily from wk 14 through 44 postpartum. Cows were fed one of two total mixed rations. Diet 1 (NE1 = 1.65 Mcal/kg, CP = 18%, and ADF = 22%) was fed from start of lactation to at least 4 wk after initiation of treatment. Cows were switched to diet 2 (NE1 = 1.56 Mcal/kg, CP = 16%, and ADF = 27%) when milk output fell below 25 kg/d. Forty-four week lactation yields were 9800 and 9447 kg milk; 364 and 354 kg fat; and 322 and 309 kg protein for high and low genetic groups, respectively. Milk, milk fat, or protein yield due to somatotropin did not differ between genetic groups. Increasing dosage of bST increased milk, 4% FCM, fat, and protein yields in a linear fashion. Percentages of fat and protein of milk were similar for all treatment groups. Body weight changes were not significantly different among treatments, but condition score changes decreased linearly with increasing dose of bST. Long-term treatment with recombinant bST had no apparent effect on incidence of health problems or reproduction.     

Lomeguatrib Increases the Radiosensitivity of MGMT Unmethylated Human Glioblastoma Multiforme Cell Lines

Background: Treatment resistance of glioblastoma multiforme to chemo- and radiotherapy remains a challenge yet to overcome. In particular, the O⁶-methylguanine-DNA-methyltransferase (MGMT) promoter unmethylated patients have only little benefit from chemotherapy treatment using temozolomide since MGMT counteracts its therapeutic efficacy. Therefore, new treatment options in radiotherapy need to be developed to inhibit MGMT and increase radiotherapy response. Methods: Lomeguatrib, a highly specific MGMT inhibitor, was used to inactivate MGMT protein in vitro. Radiosensitivity of established human glioblastoma multiforme cell lines in combination with lomeguatrib was investigated using the clonogenic survival assay. Inhibition of MGMT was analyzed using Western Blot. Cell cycle distribution and apoptosis were investigated to determine the effects of lomeguatrib alone as well as in combination with ionizing radiation. Results: Lomeguatrib significantly decreased MGMT protein and reduced radiation-induced G2/M arrest. A radiosensitizing effect of lomeguatrib was observed when administered at 1 µM and increased radioresistance at 20 µM. Conclusion: Low concentrations of lomeguatrib elicit radiosensitization, while high concentrations mediate a radioprotective effect.     

Five-year experience with setup and implementation of an integrated database system for clinical documentation and research

In radiation oncology, where treatment concepts are elaborated in interdisciplinary collaborations, handling distributed, large heterogeneous amounts of data efficiently is very important, yet challenging, for an optimal treatment of the patient as well as for research itself. This becomes a strong focus, as we step into the era of modern personalized medicine, relying on various quantitative data information, thus involving the active contribution of multiple medical specialties. Hence, combining patient data from all involved information systems is inevitable for analyses. Therefore, we introduced a documentation and data management system integrated in the clinical environment for electronic data capture. We discuss our concept and five-year experience of a precise electronic documentation system, with special focus on the challenges we encountered. We specify how such a system can be designed and implemented to plan, tailor and conduct (multicenter) clinical trials, ultimately reaching the best clinical performance, and enhancing interdisciplinary and clinical research.     

Use of a liquid chromatograph-ion trap mass spectrometer for end-group characterization of various polyethoxylate materials

The separation and detection properties of various ethoxylated polymers were investigated with atmospheric pressure ionization liquid chromatography/mass spectrometry (LC/MS). Interesting chromatographic elution behavior based on functionality was noted. LC/MS using in-source CID (collision-induced dissociation) and tandem mass spectrometry (MS/MS) detection was compared for end-group identification. Excellent end-group identification was achieved when the end-group molecular weight (MW) was greater than 100 Da and the average MW of the polymer was less than 400 Da by both MS/MS and in-source CID. In cases where the end-group MW was less than 100 Da, because of the low mass cut-off in a quadrapole ion trap analyzer, in-source CID produced the only characteristic end-group ions. The use of a dual scan function allowed investigation of the oligomeric distribution followed by a narrow low-mass scan using in-source CID to generate end-group information. This approach is applicable on instruments without MS/MS capability.     

Effects of molecular weight distribution and branching on rheological properties of polyolefin melts

The objective of this work was to determine the relationships among molecular and melt parameters of polyolefins. The polyolefins studied are polypropylene, poly-1-butene, poly-1-hexene, poly-1-dodecene, these have regularly spaced short-chain branches. Conclusions from previous work, as well as some new data, on polyethylene are given. As the molecular weight increases, the critical shear rate decreases but the melt viscosity and non-Newtonian ratio increase. As the molecular weight distribution broadens, the critical shear rate decreases, whereas the normal forces and the non-Newtonian ratio increase. Increasing the number of short-chain branches increases the energy of activation and the melt viscosity but decreases the non-Newtonian ratio. As the length of the short-chain branches increases, the non-Newtonian ratio increases, but the melt viscosity, critical shear rate, and energy of activation decrease. Increasing the number of long-chain branches decreases the non-Newtonian ratio, but the normal forces and the melt viscosity increase. Such information allows the polymer chemist to design a polyolefin molecule having the critical melt properties required for a given production technique.     

On the Reliability of a Worst Case Designed Nonredundant Circuit

A worst case designed circuit is defined and, based on observed component failure modes, a mathematical model for circuit reliability is obtained. A special case of the general model is then considered by assuming all the component failure rates to be equal. Using this simplified model it is established that a worst case designed nonredundant circuit is more reliable than the product of the reliabilities of its components. The mean time to failure (MTTF) is computed and it is shown that, for particular extreme values of the parameter p, the MTTF assumes the classical forms more frequently seen in the literature.