Detection and localization of a lectin on Actinomyces viscosus T14V by monoclonal antibodies

A cell-associated lectin activity that mediates lactose-inhibitable adherence of Actinomyces viscosus T14V has been localized to a specific population of fimbriae by the use of monoclonal antibodies. Nine monoclonal antibodies were produced that reacted with only 1 of 2 immunoelectrophoretically distinct fimbrial components on T14V. The fibrillar morphology of this component was revealed by the immunoelectronmicroscopic examination of bacteria incubated with the monoclonal antibodies. The lectin activity associated with these structures was detected when isolated fimbriae were cross-linked with monoclonal antibodies to form immune complexes with agglutination activity for neuraminidase-treated human erythrocytes, a reaction that was inhibited by lactose. Although the 9 monoclonal antibodies differed in their fine specificities, they reacted only with strains of A. viscosus and A. naeslundii that exhibited lactose-inhibitable adherence. These findings indicate that the lectin activity common to these bacteria resides on fimbriae that are antigenically related to those of T14V.     

Effects of caffeine-pretreatment on SCE and chromosome aberrations induced by monofunctional- and bifunctional-mitomycin C in bloom syndrome lymphocytes

Bloom syndrome (BS) lymphocytes, which are characterized by a high incidence (75.4 per cell) of SCE, were treated with caffeine (CAF) during the first cell cycle and with monofunctional-(M-MC) and bifunctional-(MC)mitomycin C during the second cycle. The effect on the SCE level was synergistic. The CAF-pretreated cells in combination with M-MC and MC post-treatments, had significantly higher (SCE values 152.5 and 167.9 SCE per cell, resp.) than those treated with M-MC or MC alone during the second cycle (101.1 and 116.4 SCE per cell, resp.). M-MC and MC in the presence of BrdU (without CAF) for 2 cell cycles increased SCE to 157.6 and 169.4 per cell (about twice the control level). M-MC + CAF and MC + CAF treatments for 2 cell cycles did not produce a synergistic effect on the SCE frequency in BS cells; the SCE level was not significantly greater than that with M-MC or MC alone. Normal cells treated with MC and CAF for 2 cycles had a maximum SCE frequency of 156 per cell. This suggests that cells with SCE frequencies above this level may not be able to survive, i.e., this is the “saturation” level of SCE. However, CAF alone had almost no effect on SCE in either BS or normal cells and did not produce multiple chromosome aberrations. The lack of CAF effect on BS cells suggests that the lesions in DNA strands of BS cells which lead to SCE are double-strand lesions. In normal cells CAF is known to significantly slow down DNA-chain growth; the reduced rate of DNA-chain growth in BS is an inherent defect of the cells. Therefore, though CAF enhanced SCE and chromosome aberrations (shattered chromosomes) in combination with alkylating agents, CAF alone did not significantly increase the SCE rate in either BS cells or in normal cells. Thus, processes which may induce SCE are not only related to retarded rate of DNA-chain growth, but also to breaks in the template strand permitting double-strand exchanges to occur.     

Clones of Friend leukemia cells: differences in karyotypes and responsiveness to inducers of differentiation.

Several clones of Friend leukemia cells have been established which differ in their chromosome composition. These cells also vary with regard to their responsiveness to DMSO, whereas all are responsive to butyric acid. Hence, there appears to be independent assortment of the ability of a cell line to respond to DMSO and to butyric acid, suggesting a different mechanism of action for each agent. Further, individual Friend cells possess the ability to simultaneously contain chloroacetate esterase and heme--two biochemical properties which have previously been believed to be mutually exclusive.     

The smooth muscle cell. III. Elastin synthesis in arterial smooth muscle cell culture

Primate arterial smooth muscle cells and skin fibroblasts were examined for their ability to synthesize elastin in culture. In the presence of the lathyrogen beta-aminopropionitrile, the smooth muscle cells incorporate [3H]lysine into a lysyl oxidase substrate that was present in the medium and associated with the cell layer. A component having a mol wt of 72,000 and an electrophoretic mobility similar to that of authentic tropoelastin was isolated from the labeled smooth muscle cells by coacervation and fractionation with organic solvents. In the absence of beta-aminopropionitrile, long-term cultures of smooth muscle cells incorporated [14C]lysine into desmosine and isodesmosine, the cross-link amino acids unique to elastin. In contrast, no desmosine formation occurred in the fibroblast cultures. These characteristics demonstrate that arterial smooth muscle cells are capable of synthesizing both soluble and cross-lined elastin in culture.     

Analysis of 3-indole carboxylic acid in Pinus sylvestris needles

3-Indole carboxylic acid (ICA) has been characterized as an endogenous constituent of Pinus sylvestris needles. Quantitative estimates of 3-indole acetic acid (IAA) and ICA, corrected for both sample losses and the conversion of IAA to ICA occurring during purification, indicate that Pinus needles contain 24.5 ± 6.5 ng IAA/g and 2.3 ± 0.4 ng ICA/g.     

A computational study of the EN 1078 impact test for bicycle helmets using a realistic subject-specific finite element head model

Abstract

In the present study, the free fall impact test in accordance with the EN1078 standard for certification of bicycle helmets is replicated using numerical simulations. The impact scenario is simulated using an experimentally validated, patient-specific head model equipped with and without a bicycle helmet. Head accelerations and intracranial biomechanical injury metrics during the impacts are recorded. It is demonstrated that wearing the bicycle helmet during the impact reduces biomechanical injury metrics, with the biggest reduction seen in the metric for skull fracture.     

Amyloid-β Protofibrils: Size,Morphology and Synaptotoxicity of an Engineered Mimic

Structural and biochemical studies of the aggregation of the amyloid-β peptide (Aβ) are important to understand the mechanisms of Alzheimer''s disease, but research is complicated by aggregate inhomogeneity and instability. We previously engineered a hairpin form of Aβ called Aβcc, which forms stable protofibrils that do not convert into amyloid fibrils. Here we provide a detailed characterization of Aβ₄₂ cc protofibrils. Like wild type Aβ they appear as smooth rod-like particles with a diameter of 3.1 (±0.2) nm and typical lengths in the range 60 to 220 nm when observed by atomic force microscopy. Non-perturbing analytical ultracentrifugation and nanoparticle tracking analyses are consistent with such rod-like protofibrils. Aβ₄₂ cc protofibrils bind the ANS dye indicating that they, like other toxic protein aggregates, expose hydrophobic surface. Assays with the OC/A11 pair of oligomer specific antibodies put Aβ₄₂ cc protofibrils into the same class of species as fibrillar oligomers of wild type Aβ. Aβ₄₂ cc protofibrils may be used to extract binding proteins in biological fluids and apolipoprotein E is readily detected as a binder in human serum. Finally, Aβ₄₂ cc protofibrils act to attenuate spontaneous synaptic activity in mouse hippocampal neurons. The experiments indicate considerable structural and chemical similarities between protofibrils formed by Aβ₄₂ cc and aggregates of wild type Aβ₄₂. We suggest that Aβ₄₂ cc protofibrils may be used in research and applications that require stable preparations of protofibrillar Aβ.     

Therapeutic vaccination against autologous cancer stem cells with mRNA-transfected dendritic cells in patients with glioblastoma

Background

The growth and recurrence of several cancers appear to be driven by a population of cancer stem cells (CSCs). Glioblastoma, the most common primary brain tumor, is invariably fatal, with a median survival of approximately 1 year. Although experimental data have suggested the importance of CSCs, few data exist regarding the potential relevance and importance of these cells in a clinical setting.

Methods

We here present the first seven patients treated with a dendritic cell (DC)-based vaccine targeting CSCs in a solid tumor. Brain tumor biopsies were dissociated into single-cell suspensions, and autologous CSCs were expanded in vitro as tumorspheres. From these, CSC-mRNA was amplified and transfected into monocyte-derived autologous DCs. The DCs were aliquoted to 9–18 vaccines containing 10⁷ cells each. These vaccines were injected intradermally at specified intervals after the patients had received a standard 6-week course of post-operative radio-chemotherapy. The study was registered with the ClinicalTrials.gov identifier NCT00846456.

Results

Autologous CSC cultures were established from ten out of eleven tumors. High-quality RNA was isolated, and mRNA was amplified in all cases. Seven patients were able to be weaned from corticosteroids to receive DC immunotherapy. An immune response induced by vaccination was identified in all seven patients. No patients developed adverse autoimmune events or other side effects. Compared to matched controls, progression-free survival was 2.9 times longer in vaccinated patients (median 694 vs. 236 days, p = 0.0018, log-rank test).

Conclusion

These findings suggest that vaccination against glioblastoma stem cells is safe, well-tolerated, and may prolong progression-free survival.     

Atrioventricular nodal function during atrial fibrillation: Model building and robust estimation

Statistical modeling of atrioventricular (AV) nodal function during atrial fibrillation (AF) is revisited for the purpose of defining model properties and improving parameter estimation. The characterization of AV nodal pathways is made more detailed and the number of pathways is now determined by the Bayesian information criterion, rather than just producing a probability as was previously done. Robust estimation of the shorter refractory period (i.e., of the slow pathway) is accomplished by a Hough-based technique which is applied to a Poincaré plot of RR intervals. The performance is evaluated on simulated data as well as on ECG data acquired from AF patients during rest and head-up tilt test. The simulation results suggest that the refractory period of the slow pathway can be accurately estimated even in the presence of many artifacts. They also show that the number of pathways can be accurately determined. The results from ECG data show that the refined AV node model provides significantly better fit than did the original model, increasing from 85 ± 5% to 88 ± 4% during rest, and from 86 ± 5% to 87 ± 3% during tilt. When assessing the effect of sympathetic stimulation, the AF frequency increased significantly during tilt (6.25 ± 0.58 Hz vs. 6.32 ± 0.61 Hz, p < 0.05, rest vs. tilt) and the prolongation of the refractory periods of both pathways decreased significantly (slow pathway: 0.23 ± 0.20 s vs. 0.11 ± 0.10 s, p < 0.001, rest vs. tilt; fast pathway: 0.24 ± 0.31 s vs. 0.16 ± 0.19 s, p < 0.05, rest vs. tilt). The results show that AV node characteristics can be assessed noninvasively for the purpose of quantifying changes induced by autonomic stimulation.     

Origin of the S‐Shaped JV Curve and the Light‐Soaking Issue in Inverted Organic Solar Cells

Inverted organic solar cells generally exhibit a strong s‐shaped kink in the current–voltage characteristics (JV curve) that may be removed by exposure to UV light (light‐soaking) leading to a drastically improved performance. Using in‐device characterization methods the origin of the light‐soaking issue in inverted solar cells employing titanium dioxide (TiO₂) as an electron selective layer is clarified. An injected hole reservoir accumulated at the TiO₂/organic interface of the pristine device is observed from extraction current transients; the hole reservoir increases the recombination and results in an s‐shape in the JV curve of pristine devices. The hole reservoir and the s‐shape is a result of the energetics at the selective contact in the pristine device; the effect of UV exposure is to decrease the work function of the indium tin oxide/TiO₂‐contact, increasing the built‐in potential. This hinders the build‐up of the hole reservoir and the s‐shape is removed. The proposed model is in excellent agreement with drift‐diffusion simulations.