Functional Analysis of the Hsp93/ClpC Chaperone at the Chloroplast Envelope.

The Hsp100-type chaperone Hsp93/ClpC has crucial roles in chloroplast biogenesis. In addition to its role in proteolysis in the stroma, biochemical and genetic evidence led to the hypothesis that this chaperone collaborates with the inner envelope TIC complex to power preprotein import. Recently, it was suggested that Hsp93, working together with the Clp proteolytic core, can confer a protein quality control mechanism at the envelope. Thus, the role of envelope-localized Hsp93, and the mechanism by which it participates in protein import, remain unclear. To analyze the function of Hsp93 in protein import independently of its ClpP association, we created a mutant of Hsp93 affecting its ClpP-binding motif (PBM) (Hsp93[P-]), which is essential for the chaperone's interaction with the Clp proteolytic core. The Hsp93[P-] construct was ineffective at complementing the pale-yellow phenotype of hsp93 Arabidopsis (Arabidopsis thaliana) mutants, indicating that the PBM is essential for Hsp93 function. As expected, the PBM mutation negatively affected the degradation activity of the stromal Clp protease. The mutation also disrupted association of Hsp93 with the Clp proteolytic core at the envelope, without affecting the envelope localization of Hsp93 itself or its association with the TIC machinery, which we demonstrate to be mediated by a direct interaction with Tic110. Nonetheless, Hsp93[P-] expression did not detectably improve the protein import efficiency of hsp93 mutant chloroplasts. Thus, our results do not support the proposed function of Hsp93 in protein import propulsion, but are more consistent with the notion of Hsp93 performing a quality control role at the point of import.

Application of Theranostics in Oncology.

In recent years, due to the development of nanotechnology new horizons in treatment and diagnosis of cancer open up. Development of nano-systems for simultaneous transfer of active substances and imaging of tumor regions gathers an important amount of scientific interest. This new category of nano-systems is called Theranostics. Theranostics methods can provide multiple benefits by inserting nanoparticles into the patient and using photodynamic therapy and pave the way for personalized medicine. The objective of this paper is to study the use and application of Theranostics in the diagnosis and treatment of cancer, in order to achieve personalized anticancer treatment. For this purpose, investigation of existing literature has been conducted using electronic databases, PubMed, Google Scholar and IEEE Xplore. In addition, there was a secondary research phase, using paper citations found during the first research phase. It has to be pointed out that nanoparticles are the basis of Theranostics, since, due to their properties, they provide the ability to display accurate imaging and provide diagnosis along with simultaneous treatment of diseases. Theranostics methods may be applied in treatment of esophageal cancer, prostate cancer, breast cancer, in treatment of actinic keratosis, actinic cheilitis and Bowen's disease and in treatment of basal cell epithelioma and macular degeneration. As a result, application of Theranostics can provide multiple benefits by inserting nanoparticles into the patient. This method is currently encountering many challenges, but continuation of research on the field is necessary not only for the improvement of the medical field and the healthcare techniques, but also for the creation of new treatment methods for patients with diseases that are incurable until now.

Applications for Electrical Impedance Tomography (EIT) and Electrical Properties of the Human Body.

Electrical Impedance Tomography (EIT) is a promising application that displays changes in conductivity within a body. The basic principle of the method is the repeated measurement of surface voltages of a body, which are a result of rolling injection of known and small-volume sinusoidal AC current to the body through the electrodes attached to its surface. This method finds application in biomedicine, biology and geology. The objective of this paper is to present the applications of Electrical Impedance Tomography, along with the method's capabilities and limitations due to the electrical properties of the human body. For this purpose, investigation of existing literature has been conducted, using electronic databases, PubMed, Google Scholar and IEEE Xplore. In addition, there was a secondary research phase, using paper citations found during the first research phase. It should be noted that Electrical Impedance Tomography finds use in a plethora of medical applications, as the different tissues of the body have different conductivities and dielectric constants. Main applications of EIT include imaging of lung function, diagnosis of pulmonary embolism, detection of tumors in the chest area and diagnosis and distinction of ischemic and hemorrhagic stroke. EIT advantages include portability, low cost and safety, which the method provide, since it is a noninvasive imaging method that does not cause damage to the body. The main disadvantage of the method, which blocks its wider spread, appears in the image composition from the voltage measurements, which are conducted by electrodes placed on the periphery of the body, because the injected currents are affected nonlinearly by the general distribution of the electrical properties of the body. Furthermore, the complex impedance of the skin-electrode interface can be modelled by using a capacitor and two resistor, as a result of skin properties. In conclusion, Electrical Impedance Tomography is a promising method for the development of noninvasive diagnostic medicine, since it is able to provide imaging of the interior of the human body in real time without causing harm or putting the human body in risk.

Quantitative analysis of the chloroplast molecular chaperone ClpC/Hsp93 in Arabidopsis reveals new insights into its localization, interaction with the Clp proteolytic core, and functional importance.

The molecular chaperone ClpC/Hsp93 is essential for chloroplast function in vascular plants. ClpC has long been held to act both independently and as the regulatory partner for the ATP-dependent Clp protease, and yet this and many other important characteristics remain unclear. In this study, we reveal that of the two near-identical ClpC paralogs (ClpC1 and ClpC2) in Arabidopsis chloroplasts, along with the closely related ClpD, it is ClpC1 that is the most abundant throughout leaf maturation. An unexpectedly large proportion of both chloroplast ClpC proteins (30% of total ClpC content) associates to envelope membranes in addition to their stromal localization. The Clp proteolytic core is also bound to envelope membranes, the amount of which is sufficient to bind to all the similarly localized ClpC. The role of such an envelope membrane Clp protease remains unclear although it appears uninvolved in preprotein processing or Tic subunit protein turnover. Within the stroma, the amount of oligomeric ClpC protein is less than that of the Clp proteolytic core, suggesting most if not all stromal ClpC functions as part of the Clp protease; a proposal supported by the near abolition of Clp degradation activity in the clpC1 knock-out mutant. Overall, ClpC appears to function primarily within the Clp protease, as the principle stromal protease responsible for maintaining homeostasis, and also on the envelope membrane where it possibly confers a novel protein quality control mechanism for chloroplast preprotein import.

Branching principles of animal and plant networks identified by combining extensive data, machine learning and modelling.

Branching in vascular networks and in overall organismic form is one of the most common and ancient features of multicellular plants, fungi and animals. By combining machine-learning techniques with new theory that relates vascular form to metabolic function, we enable novel classification of diverse branching networks-mouse lung, human head and torso, angiosperm and gymnosperm plants. We find that ratios of limb radii-which dictate essential biologic functions related to resource transport and supply-are best at distinguishing branching networks. We also show how variation in vascular and branching geometry persists despite observing a convergent relationship across organisms for how metabolic rate depends on body mass.

Ubiquitin-dependent chloroplast-associated protein degradation in plants.

Chloroplasts contain thousands of nucleus-encoded proteins that are imported from the cytosol by translocases in the chloroplast envelope membranes. Proteolytic regulation of the translocases is critically important, but little is known about the underlying mechanisms. We applied forward genetics and proteomics in Arabidopsis to identify factors required for chloroplast outer envelope membrane (OEM) protein degradation. We identified SP2, an Omp85-type ß-barrel channel of the OEM, and CDC48, a cytosolic AAA+ (ATPase associated with diverse cellular activities) chaperone. Both proteins acted in the same pathway as the ubiquitin E3 ligase SP1, which regulates OEM translocase components. SP2 and CDC48 cooperated to bring about retrotranslocation of ubiquitinated substrates from the OEM (fulfilling conductance and motor functions, respectively), enabling degradation of the substrates by the 26S proteasome in the cytosol. Such chloroplast-associated protein degradation (CHLORAD) is vital for organellar functions and plant development.

Phytochrome and Phytohormones: Working in Tandem for Plant Growth and Development.

Being sessile organisms, plants need to continually adapt and modulate their rate of growth and development in accordance with the changing environmental conditions, a phenomenon referred to as plasticity. Plasticity in plants is a highly complex process that involves a well-coordinated interaction between different signaling pathways, the spatiotemporal involvement of phytohormones and cues from the environment. Though research studies are being carried out over the years to understand how plants perceive the signals from changing environmental conditions and activate plasticity, such remain a mystery to be resolved. Among all environmental cues, the light seems to be the stand out factor influencing plant growth and development. During the course of evolution, plants have developed well-equipped signaling system that enables regulation of both quantitative and qualitative differences in the amount of perceived light. Light influences essential developmental switches in plants ranging from germination or transition to flowering, photomorphogenesis, as well as switches in response to shade avoidances and architectural changes occurring during phototropism. Abscisic acid (ABA) is controlling seed germination and is regulated by light. Furthermore, circadian clock adds another level of regulation to plant growth by integrating light signals with different hormonal pathways. MYB96 has been identified as a regulator of circadian gating of ABA-mediated responses in plants by binding to the TIMING OF CAB EXPRESSION 1(TOC1) promoter. This review will present a representative regulatory model, highlight the successes achieved in employing novel strategies to dissect the levels of interaction and provide perspective for future research on phytochrome-phytohormones relationships toward facilitating plant growth, development, and function under abiotic-biotic stresses.

Computational study of the influence of callus porosity on ultrasound propagation in healing bones.

In the process of fracture healing, several phases of recovery are observed as the mechanical stability, continuity and normal load carrying capacity are gradually restored. The ultrasonic monitoring and discrimination of different healing stages is a complex process due to the significant microstructure and porous nature of osseous and callus tissues. In this study, we investigate the influence of the callus pores' size and concentration on ultrasound propagation in a long bone at a late healing stage. Different excitation frequencies are applied in the range of 300 kHz-1 MHz. A 2D geometry is developed and axial transmission calculations are performed based on a Finite Element Method. The velocity of the first arriving signal (FAS) and the propagation of guided waves are used as the estimated parameters. It was shown that the FAS velocity can reflect callus porosity changes, while the propagation of guided waves is sensitive to pores' distribution for higher frequencies.