Higher Derivative Fermionic Field Equation in the First Order Formalism

The generalized Dirac equation of the third order, describing particles with spin 1/2 and three mass states, is analyzed. We obtain the first order generalized Dirac equation in the 24-dimensional matrix form. The mass and spin projection operators are found which extract solutions of the wave equation corresponding to pure spin states of particles. The density of the electromagnetic current is obtained, and minimal and non-minimal (anomalous) electromagnetic interactions of fermions are considered by introducing three phenomenological parameters. The Hamiltonian form of the first order equation has been obtained.Comment: 16 pages, title changed, new section, appendixes, and references adde

Nano-Motion Analysis for Rapid and Label Free Assessing of Cancer Cell Sensitivity to Chemotherapeutics.

Background and Objectives: Optimization of chemotherapy is crucial for cancer patients. Timely and costly efficient treatments are emerging due to the increasing incidence of cancer worldwide. Here, we present a methodology of nano-motion analysis that could be developed to serve as a screening tool able to determine the best chemotherapy option for a particular patient within hours. Materials and Methods: Three different human cancer cell lines and their multidrug resistant (MDR) counterparts were analyzed with an atomic force microscope (AFM) using tipless cantilevers to adhere the cells and monitor their nano-motions. Results: The cells exposed to doxorubicin (DOX) differentially responded due to their sensitivity to this chemotherapeutic. The death of sensitive cells corresponding to the drop in signal variance occurred in less than 2 h after DOX application, while MDR cells continued to move, even showing an increase in signal variance. Conclusions: Nano-motion sensing can be developed as a screening tool that will allow simple, inexpensive and quick testing of different chemotherapeutics for each cancer patient. Further investigations on patient-derived tumor cells should confirm the method's applicability

PTPRF is disrupted in a patient with syndromic amastia

<p>Abstract</p> <p>Background</p> <p>The presence of mammary glands distinguishes mammals from other organisms. Despite significant advances in defining the signaling pathways responsible for mammary gland development in mice, our understanding of human mammary gland development remains rudimentary. Here, we identified a woman with bilateral amastia, ectodermal dysplasia and unilateral renal agenesis. She was found to have a chromosomal balanced translocation, 46,XX,t(1;20)(p34.1;q13.13). In addition to characterization of her clinical and cytogenetic features, we successfully identified the interrupted gene and studied its consequences.</p> <p>Methods</p> <p>Characterization of the breakpoints was performed by molecular cytogenetic techniques. The interrupted gene was further analyzed using quantitative real-time PCR and western blotting. Mutation analysis and high-density SNP array were carried out in order to find a pathogenic mutation. Allele segregations were obtained by haplotype analysis.</p> <p>Results</p> <p>We enabled to identify its breakpoint on chromosome 1 interrupting the <it>protein tyrosine receptor type F gene </it>(<it>PTPRF</it>). While the patient's mother and sisters also harbored the translocated chromosome, their non-translocated chromosomes 1 were different from that of the patient. Although a definite pathogenic mutation on the paternal allele could not be identified, <it>PTPRF</it>'s RNA and protein of the patient were significantly less than those of her unaffected family members.</p> <p>Conclusions</p> <p>Although <it>ptprf </it>has been shown to involve in murine mammary gland development, no evidence has incorporated <it>PTPRF </it>in human organ development. We, for the first time, demonstrated the possible association of <it>PTPRF </it>with syndromic amastia, making it a prime candidate to investigate for its spatial and temporal roles in human breast development.</p

Tubulin isoform composition tunes microtubule dynamics

Microtubules polymerize and depolymerize stochastically, a behavior essential for cell division, motility and differentiation. While many studies advanced our understanding of how microtubule-associated proteins tune microtubule dynamics in trans, we have yet to understand how tubulin genetic diversity regulates microtubule functions. The majority of in vitro dynamics studies are performed with tubulin purified from brain tissue. This preparation is not representative of tubulin found in many cell types. Here we report the 4.2Å cryo-EM structure and in vitro dynamics parameters of α1B/βI+βIVb microtubules assembled from tubulin purified from a human embryonic kidney cell line with isoform composition characteristic of fibroblasts and many immortalized cell lines. We find that these microtubules grow faster and transition to depolymerization less frequently compared to brain microtubules. Cryo-EM reveals that the dynamic ends of α1B/βI+βIVb microtubules are less tapered and that these tubulin heterodimers display lower curvatures. Interestingly, analysis of EB1 distributions at dynamic ends suggests no differences in GTP cap sizes. Lastly, we show that the addition of recombinant α1A/βIII tubulin, a neuronal isotype overexpressed in many tumors, proportionally tunes the dynamics of α1B/βI+βIVb microtubules. Our study is an important step towards understanding how tubulin isoform composition tunes microtubule dynamics

The Role of Actin Turnover in Retrograde Actin Network Flow in Neuronal Growth Cones

The balance of actin filament polymerization and depolymerization maintains a steady state network treadmill in neuronal growth cones essential for motility and guidance. Here we have investigated the connection between depolymerization and treadmilling dynamics. We show that polymerization-competent barbed ends are concentrated at the leading edge and depolymerization is distributed throughout the peripheral domain. We found a high-to-low G-actin gradient between peripheral and central domains. Inhibiting turnover with jasplakinolide collapsed this gradient and lowered leading edge barbed end density. Ultrastructural analysis showed dramatic reduction of leading edge actin filament density and filament accumulation in central regions. Live cell imaging revealed that the leading edge retracted even as retrograde actin flow rate decreased exponentially. Inhibition of myosin II activity before jasplakinolide treatment lowered baseline retrograde flow rates and prevented leading edge retraction. Myosin II activity preferentially affected filopodial bundle disassembly distinct from the global effects of jasplakinolide on network turnover. We propose that growth cone retraction following turnover inhibition resulted from the persistence of myosin II contractility even as leading edge assembly rates decreased. The buildup of actin filaments in central regions combined with monomer depletion and reduced polymerization from barbed ends suggests a mechanism for the observed exponential decay in actin retrograde flow. Our results show that growth cone motility is critically dependent on continuous disassembly of the peripheral actin network

Development of graphene-based biosensor for medical diagnostics

The explosion of information provided by the “-omics,” (genomics, proteomics, etc.) has resulted in a pressing need to develop matching diagnostic technologies, so-called biosensors. Rapid, sensitive, selective, and cost-effective analysis of different biomolecules and microorganisms is crucial in clinical diagnosis and efficient treatment of patients. Further, there is a growing demand for decentralized laboratory methodologies that can be implemented in doctor’s office, emergency room or in the field for the analysis of such analytes as DNA, RNA, proteins, antibodies, bacteria, viruses, small compounds etc. Lab-on-a-chip platforms and miniaturized point-of-care devices based on biosensors fulfill these demands and are foreseen to revolutionize the future of medical diagnostics. Because of excellent electric and optical properties, graphene has recently found to be highly attractive in biosensing applications and may thrust new possibilities into the field of miniaturized medical diagnostic devices. The main objective of this project is to develop a multifunctional grapheme biosensor for effective electrochemical detection of specific DNA microbial targets in biological samples. Novel nanocomposites consisting of chitosan and nanoparticle-modified graphene will be combined with locked nucleic acid molecular beacons with the goal of producing “ink” for ultrasonic non-contact printing of electrical circuits. The developed technology will allow fabrication of low cost, highly sensitive biosensors for point-of-care diagnosis

One-step purification of assembly-competent tubulin from diverse eukaryotic sources.

We have developed a protocol that allows rapid and efficient purification of native, active tubulin from a variety of species and tissue sources by affinity chromatography. The affinity matrix comprises a bacterially expressed, recombinant protein, the TOG1/2 domains from Saccharomyces cerevisiae Stu2, covalently coupled to a Sepharose support. The resin has a high capacity to specifically bind tubulin from clarified crude cell extracts, and, after washing, highly purified tubulin can be eluted under mild conditions. The eluted tubulin is fully functional and can be efficiently assembled into microtubules. The method eliminates the need to use heterologous systems for the study of microtubule-associated proteins and motor proteins, which has been a major issue in microtubule-related research