Ion diffusion in channels containing random arrays of microspheres: an electrochemical time-of-flight method

Rates of chloride ion diffusion in narrow (ca. 3 microm thick), rectangular (ca. 0.1 x 1.0 mm(2)) channels partially filled with polystyrene microspheres are investigated by a potentiometric electrochemical time-of-flight (P-ETOF) method. Lithographically fabricated on glass slides, P-ETOF devices consist of a centrally positioned 10 microm wide, ca. 1 mm long generator microelectrode and two sensor microelectrodes of the same dimensions symmetrically positioned on both side of the generator at a distance of 50 microm. The electrodes are silver-plated and partially oxidized in a chloride electrolyte to form Ag/AgCl deposits. Constant current reduction of AgCl on the generator electrode is used to produce chloride ions at a constant rate. Ag/AgCl deposited on the sensor microelectrodes allows time-dependent potentiometric monitoring of the increasing concentration of chloride ions diffusing across the interelectrode gap. The device is enclosed with a parallel glass plate to form a narrow channel with the polystyrene microbeads serving as spacers. The packing density of the microspheres expressed in terms of the fractional void volume (rho) varied from ca. 0.6 to 1.0. Using rho, we modified a diffusion equation describing the change of chloride ion concentration at the sensor microelectrode to include the effect of the microspheres restricting the void volume. We rely on digital simulations as well as on direct P-ETOF experiments to show that the proposed equation does accurately account for the effect of rho on the diffusion processes. We thus demonstrate that P-ETOF can be used to measure the number of identical microspheres in the active region of a narrow channel device. In the latter context, a future application of P-ETOF as a signal transduction mechanism in biosensors is outlined.     

Reasoning with first order nondeterministic specifications

The paper presents a variant of first order logic for specifying nondeterministic software. Models of the logics are multialgebras, i.e. multi-sorted algebras with set-valued operations, together with multi-sorted valuations of variables. We allow empty carrier sets but the valuations are kept total. Terms are interpreted as sets and the usual set of algebraic terms is extended by an additional construct used for limiting nondeterminism. Atomic formulae are of the form where is a rewrite operator, corresponding semantically to inclusion. For the above logic, we develop two complete deduction systems in the natural deduction style: a Rasiowa-Sikorski system for sequences of formulae, and a cut-free Gentzen-style sequent calculus. We also consider the issues of determinism and partiality, proposing alternate solutions to defining the respective predicates in our logic. Received: 4 March 1997 / 25 May 1999     

Curcumin and Its New Derivatives: Correlation between Cytotoxicity against Breast Cancer Cell Lines,Degradation of PTP1B Phosphatase and ROS Generation

Breast cancer is the most common cancer of women—it affects more than 2 million women worldwide. PTP1B phosphatase can be one of the possible targets for new drugs in breast cancer therapy. In this paper, we present new curcumin derivatives featuring a 4-piperidone ring as PTP1B inhibitors and ROS inducers. We performed cytotoxicity analysis for twelve curcumin derivatives against breast cancer MCF-7 and MDA-MB-231 cell lines and the human keratinocyte HaCaT cell line. Furthermore, because curcumin is a known antioxidant, we assessed antioxidant effects in its derivatives. For the most potent cytotoxic compounds, we determined intracellular ROS and PTP1B phosphatase levels. Moreover, for curcumin and its derivatives, we performed real-time microscopy to observe the photosensitizing effect. Finally, computational analysis was performed for the curcumin derivatives with an inhibitory effect against PTP1B phosphatase to assess the potential binding mode of new inhibitors within the allosteric site of the enzyme. We observed that two tested compounds are better anticancer agents than curcumin. Moreover, we suggest that blocking the -OH group in phenolic compounds causes an increase in the cytotoxicity effect, even at a low concentration. Furthermore, due to this modification, a higher level of ROS is induced, which correlates with a lower level of PTP1B.     

Preclinical Study of Immunological Isoxazole Derivatives as a Potential Support for Melanoma Chemotherapy

(1) Background: Melanoma is an aggressive neoplasm derived from melanocyte precursors with a high metastatic potential. Responses to chemotherapy and immunotherapy for melanoma remain weak, underlining the urgent need to develop new therapeutic strategies for the treatment of melanoma. (2) Methods: The viability of NHDF and A375 cell cultures after the administration of the tested isoxazole derivatives was assessed after 24-h and 48-h incubation periods with the test compounds in the MTT test. ROS and NO scavenging analyses, a glycoprotein-P activity analysis, a migration assay, a test of apoptosis, and a multiple-criteria decision analysis were also performed. (3) Results: All compounds that were tested resulted in a slower migration of melanoma neoplastic cells. The mechanism of the antitumor activity of the tested compounds was confirmed—i.e., the pro-apoptotic activity of the compounds in A375 cell cultures. Compound O7K qualified for further research. (4) Conclusions: All the tested compounds inhibited the formation of melanoma metastases and demonstrated the ability to reduce the risk of developing drug resistance in the tumor. The MCDA results showed that O7K showed the strongest antitumor activity.     

Novel Biomolecules in the Pathogenesis of Gestational Diabetes Mellitus

Gestational diabetes mellitus (GDM) is one of the most common metabolic diseases in pregnant women. Its early diagnosis seems to have a significant impact on the developing fetus, the course of delivery, and the neonatal period. It may also affect the later stages of child development and subsequent complications in the mother. Therefore, the crux of the matter is to find a biopredictor capable of singling out women at risk of developing GDM as early as the very start of pregnancy. Apart from the well-known molecules with a proven and clear-cut role in the pathogenesis of GDM, e.g., adiponectin and leptin, a potential role of newer biomolecules is also emphasized. Less popular and less known factors with different mechanisms of action include: galectins, growth differentiation factor-15, chemerin, omentin-1, osteocalcin, resistin, visfatin, vaspin, irisin, apelin, fatty acid-binding protein 4 (FABP4), fibroblast growth factor 21, and lipocalin-2. The aim of this review is to present the potential and significance of these 13 less known biomolecules in the pathogenesis of GDM. It seems that high levels of FABP4, low levels of irisin, and high levels of under-carboxylated osteocalcin in the serum of pregnant women can be used as predictive markers in the diagnosis of GDM. Hopefully, future clinical trials will be able to determine which biomolecules have the most potential to predict GDM.     

Species Phylogeny versus Gene Trees: A Case Study of an Incongruent Data Matrix Based on Paphiopedilum Pfitz. (Orchidaceae)

The phylogeny of the genus Paphiopedilum based on the plastome is consistent with morphological analysis. However, to date, none of the analyzed nuclear markers has confirmed this. Topology incongruence among the trees of different nuclear markers concerns entire sections of the subgenus Paphiopedilum. The low-copy nuclear protein-coding gene PHYC was obtained for 22 species representing all sections and subgenera of Paphiopedilum. The nuclear-based phylogeny is supported by morphological characteristics and plastid data analysis. We assumed that an incongruence in nuclear gene trees is caused by ancestral homoploid hybridization. We present a model for inferring the phylogeny of the species despite the incongruence of the different tree topologies. Our analysis, based on six low-copy nuclear genes, is congruent with plastome phylogeny and has been confirmed by phylogenetic network analysis.     

Tks5 Regulates Synaptic Podosome Formation and Stabilization of the Postsynaptic Machinery at the Neuromuscular Junction

Currently, the etiology of many neuromuscular disorders remains unknown. Many of them are characterized by aberrations in the maturation of the neuromuscular junction (NMJ) postsynaptic machinery. Unfortunately, the molecular factors involved in this process are still largely unknown, which poses a great challenge for identifying potential therapeutic targets. Here, we identified Tks5 as a novel interactor of αdystrobrevin-1, which is a crucial component of the NMJ postsynaptic machinery. Tks5 has been previously shown in cancer cells to be an important regulator of actin-rich structures known as invadosomes. However, a role of this scaffold protein at a synapse has never been studied. We show that Tks5 is crucial for remodeling of the NMJ postsynaptic machinery by regulating the organization of structures similar to the invadosomes, known as synaptic podosomes. Additionally, it is involved in the maintenance of the integrity of acetylcholine receptor (AChR) clusters and regulation of their turnover. Lastly, our data indicate that these Tks5 functions may be mediated by its involvement in recruitment of actin filaments to the postsynaptic machinery. Collectively, we show for the first time that the Tks5 protein is involved in regulation of the postsynaptic machinery.