Cathepsin L2, a novel human cysteine proteinase produced by breast and colorectal carcinomas

We have identified and cloned a new member of the papain family of cysteine proteinases from a human brain cDNA library. The isolated cDNA codes for a polypeptide of 334 amino acids that exhibits all of the structural features characteristic of cysteine proteinases, including the active site cysteine residue essential for peptide hydrolysis. Pairwise comparisons of this amino acid sequence with the remaining human cysteine proteinases identified to date showed a high percentage of identity (78%) with cathepsin L; the percentage of identity with all other members of the family was much lower (<40%). On the basis of these structural characteristics, we have tentatively called this novel protein cathepsin L2. The cDNA encoding the mature cathepsin L2 was expressed in Escherichia coli, and after purification, the recombinant protein was able to degrade the synthetic peptide benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methylcoumarin, which is commonly used as a substrate for cysteine proteinases. Cathepsin L2 proteolytic activity on this substrate was abolished by trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, an inhibitor of cysteine proteinases, thus providing additional evidence that the isolated cDNA encodes a functional cysteine proteinase. Northern blot analysis of polyadenylated RNAs isolated from a variety of human tissues demonstrated that cathepsin L2 is predominantly expressed in the thymus and testis. This finding is in marked contrast with the wide tissue distribution of most cysteine proteinases characterized to date, including cathepsin L, and suggests that cathepsin L2 may play a specialized role in the thymus and testis. Expression analysis of cathepsin L2 in human tumors revealed a widespread expression in colorectal and breast carcinomas but not in normal colon or mammary gland or in peritumoral tissues. Cathepsin L2 was also expressed by colorectal and breast cancer cell lines as well as by some tumors of diverse origin, including ovarian and renal carcinomas. These results open the possibility that this novel enzyme may be involved in tumor processes, as already reported for other cysteine proteinases, including cathepsin L.     

Bone growth and bone development in the presence of implants or after induced leg-lengthening studied using the Oxford Scanning Proton Microprobe

To respond to varying environmental demands the bone tissue in the body is under continual reconstruction throughout life. It is known that metallic elements are important for maintaining normal bone structure, but their roles are not well understood. More information about the effects of metal excess or deficiency is needed to help in the development of metallic bone implants and to improve the treatment of bone fractures and defects. The Oxford Scanning Proton Microprobe (SPM) is being applied in two studies involving metal ions in bone: (1) bone regrowth and bonding to titanium bone implants may be influenced by diffusion of Ti ions into the bone. We are using microPIXE to determine the metal ion content of bone developing in contact with implants of pure Nb, Ti and Ti alloys. (2) Bone lengthening as a surgical procedure is induced by fracturing the bone and allowing it to heal with a small gap between the fractured ends created by the use of external fixators. The gap can be slowly increased during the healing process to stimulate the production of new bone. The enzymes and other constituents of the developing bone need certain metals for their function. Using experimental animals we have studied the concentrations of the metals and whether a deficiency of trace metals limits the optimum rate of bone lengthening.     

Creativity in non‐routine jobs: The role of transformational leadership and organizational identification

An increasing number of individuals work in jobs with little standardization and repetition, that is, with high levels of job non‐routinization. At the same time, demands for creativity are high, which raises the question of how employees can use job non‐routinization to develop creativity. Acknowledging the importance of social processes for creativity, we propose that transformational leaders raise feelings of organizational identification in followers and that this form of identification then helps individuals to develop creativity in jobs with little routinization. This is because organizational members evaluate and promote those ideas as more creative, which are in line with a shared understanding of creativity within the organization. To investigate these relationships, we calculated a mediated moderation model with 173 leader–follower dyads from China. Results confirm our hypotheses that transformational leadership moderates the relationship between job non‐routinization on employee creativity through organizational identification. We conclude that raising feelings of social identity is a key task for leaders today, especially when working in uncertain and fast developing environments with little repetition and the constant need to develop creative ideas.     

The role of visual complexity and prototypicality regarding first impression of websites: Working towards understanding aesthetic judgments

This paper experimentally investigates the role of visual complexity (VC) and prototypicality (PT) as design factors of websites, shaping users' first impressions by means of two studies. In the first study, 119 screenshots of real websites varying in VC (low vs. medium vs. high) and PT (low vs. high) were rated on perceived aesthetics. Screenshot presentation time was varied as a between-subject factor (50 ms vs. 500 ms vs. 1000 ms). Results reveal that VC and PT affect participants' aesthetics ratings within the first 50 ms of exposure. In the second study presentation times were shortened to 17, 33 and 50 ms. Results suggest that VC and PT affect aesthetic perception even within 17 ms, though the effect of PT is less pronounced than the one of VC. With increasing presentation time the effect of PT becomes as influential as the VC effect. This supports the reasoning of the information-processing stage model of aesthetic processing (Leder et al., 2004), where VC is processed at an earlier stage than PT. Overall, websites with low VC and high PT were perceived as highly appealing.     

A system-on-chip development of a neuro-fuzzy embedded agent for ambient-intelligence environments

This paper presents the development of a neuro-fuzzy agent for ambient-intelligence environments. The agent has been implemented as a system-on-chip (SoC) on a reconfigurable device, i.e., a field-programmable gate array. It is a hardware/software (HW/SW) architecture developed around a MicroBlaze processor (SW partition) and a set of parallel intellectual property cores for neuro-fuzzy modeling (HW partition). The SoC is an autonomous electronic device able to perform real-time control of the environment in a personalized and adaptive way, anticipating the desires and needs of its inhabitants. The scheme used to model the intelligent agent is a particular class of an adaptive neuro-fuzzy inference system with piecewise multilinear behavior. The main characteristics of our model are computational efficiency, scalability, and universal approximation capability. Several online experiments have been performed with data obtained in a real ubiquitous computing environment test bed. Results obtained show that the SoC is able to provide high-performance control and adaptation in a life-long mode while retaining the modeling capabilities of similar agent-based approaches implemented on larger computing machines.     

Adenovirus-Mediated Inducible Expression of a PD-L1 Blocking Antibody in Combination with Macrophage Depletion Improves Survival in a Mouse Model of Peritoneal Carcinomatosis

Immune checkpoint inhibitors (ICIs) have demonstrated remarkable efficacy in a growing number of malignancies. However, overcoming primary or secondary resistances is difficult due to pharmacokinetics issues and side effects associated with high systemic exposure. Local or regional expression of monoclonal antibodies (mAbs) using gene therapy vectors can alleviate this problem. In this work, we describe a high-capacity adenoviral vector (HCA-EFZP-aPDL1) equipped with a mifepristone-inducible system for the controlled expression of an anti-programmed death ligand 1 (PD-L1) blocking antibody. The vector was tested in an immune-competent mouse model of colorectal cancer based on implantation of MC38 cells. A single local administration of HCA-EFZP-aPDL1 in subcutaneous lesions led to a significant reduction in tumor growth with minimal release of the antibody in the circulation. When the vector was tested in a more stringent setting (rapidly progressing peritoneal carcinomatosis), the antitumor effect was marginal even in combination with other immune-stimulatory agents such as polyinosinic-polycytidylic acid (pI:C), blocking mAbs for T cell immunoglobulin, mucin-domain containing-3 (TIM-3) or agonistic mAbs for 4-1BB (CD137). In contrast, macrophage depletion by clodronate liposomes enhanced the efficacy of HCA-EFZP-aPDL1. These results highlight the importance of addressing macrophage-associated immunoregulatory mechanisms to overcome resistance to ICIs in the context of colorectal cancer.     

Gas hold-up, mixing time and gas-liquid volumetric mass transfer coefficient of various multiple-impeller configurations: Rushton turbine, pitched blade and techmix impeller and their combinations

Gas hold-up, mixing intensity of dispersion characterised by exchange flows between adjacent impellers and a volumetric mass transfer coefficient are presented for 18 impeller configurations in triple-impeller vessel of inner diameter . Rushton Turbines, six Pitched Blade impellers pumping down and hydrofoil impellers Techmix 335 (Techmix co., Czech republic) pumping up or down and their combinations were used. aqueous solution was used as a liquid phase, which represents non-coalescent batches. Gas hold-ups and volumetric mass transfer coefficients are presented for individual configurations as functions of specific power dissipated and superficial gas velocity. The regression of the mass transfer coefficients shows large standard deviation (30%). The power number included to the regression to express the impeller configuration effect did not improve the standard deviation significantly (23%). The impeller configurations with low power number (less than unity) provide higher dispersion mixing intensities, while the impeller configurations with high power number provide better mass transfer performance.     

Poly(p-tert-butoxycarbonyloxystyrene): a convenient precursor to p-hydroxystyrene resins

An efficient synthetic route to pure, high molecular weight poly(p-hydroxystyrene) is reported. The route involves synthesis of a new monomer, p-tert-butoxycarbonyloxystyrene, polymerization by radical initiation or by cationic initiation in liquid SO₂, followed by thermolysis or acidolysis of the tert-butoxycarbonyl protecting group. Porous, crosslinked resin beads containing the nucleophilic, phenol pendant group have been prepared in a similar fashion from the precursor terpolymer of p-tert-butoxycarbonyloxystyrene, styrene and divinylbenzene. The utility of this resin for solid-phase synthesis has been demonstrated.     

A study of sulfur bridges in filled natural rubber vulcanizates by pyrolysis gas chromatography with flame-photometric detection

Natural rubber vulcanizates of two formulations, with sulfur/N-cyclohexyl-2-benzothiazolesulfenamide (CBS) and tetramethylthiuram disulfide (TMTD), respectively, were analyzed by pyrolysis gas chromatography. A sulfur-selective flame-photometric detector was utilized. The main pyrolysis products were identified as CS₂ and some thiophenes. The yields of the pyrolysis products from the two types of rubber were very different. The yields also varied with the curing time of the rubbers.     

Sufficient conditions for stability and stabilization of networked control systems with uncertainties and nonlinearities

In this paper, we consider the problem of robust stability and stabilization for networked control systems (NCS) with uncertain/nonlinear dynamics AUTHOR: Please check that authors and their affiliations are correct. in which the network‐induced delays are time‐varying and bounded. Based on some recent achievements, a relatively simple Lyapunov–Krasovskii functional is proposed to derive sufficient conditions both for analysis and synthesis of NCS in the form of LMIs depending on the delay bounds. The effectiveness of the proposed method is illustrated by several benchmark examples available in the literature. Copyright © 2014 John Wiley & Sons, Ltd.