Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

Monoclonal and recombinant antibodies are widely used for the diagnostics and therapy of cancer. They are generated to interact with cell surface proteins which are usually involved in the development and progression of cancer. Carbonic anhydrase XII (CA XII) contributes to the survival of tumors under hypoxic conditions thus is considered a candidate target for antibody-based therapy. In this study, we have generated a novel collection of monoclonal antibodies (MAbs) against the recombinant extracellular domain of CA XII produced in HEK-293 cells. Eighteen out of 24 MAbs were reactive with cellular CA XII on the surface of live kidney and lung cancer cells as determined by flow cytometry. One MAb 14D6 also inhibited the enzymatic activity of recombinant CA XII as measured by the stopped-flow assay. MAb 14D6 showed the migrastatic effect on human lung carcinoma A549 and renal carcinoma A498 cell lines in a ‘wound healing’ assay. It did not reduce the growth of multicellular lung and renal cancer spheroids but reduced the cell viability by the ATP Bioluminescence assay. Epitope mapping revealed the surface-exposed amino acid sequence (35-FGPDGENS-42) close to the catalytic center of CA XII recognized by the MAb 14D6. The variable regions of the heavy and light chains of MAb 14D6 were sequenced and their complementarity-determining regions were defined. The obtained variable sequences were used to generate recombinant antibodies in two formats: single-chain fragment variable (scFv) expressed in E. coli and scFv fused to human IgG1 Fc fragment (scFv-Fc) expressed in Chinese Hamster Ovary (CHO) cells. Both recombinant antibodies maintained the same specificity for CA XII as the parental MAb 14D6. The novel antibodies may represent promising tools for CA XII-related cancer research and immunotherapy.     

Autosensitized Photooxidation of the Rose Hip (Wild Rose) Oil - Mechanism and Quantum Yield for Singlet Molecular Oxygen O2 (1 Δg) Generation

The autosensitized photooxidation of the fatty acid components of rose hip (Wild Rose) oil was studied. Two important facts arise from the present investigation: a) Fatty acids of rose hip oil decompose upon visible light irradiation in the absence of external sensitizers by a mechanism that involves O₂(¹Δ_g). The pigments present in rose hip oil themselves efficiently sensitize the generation of O₂(¹Δ_g) with a quantum yield of 0.038 (upper limit). b) Visible irradiation also produces the degradation of the anthocyanines, native pigments which could behave as O₂(¹Δ_g) sensitizers in the oil. In this case a mechanism involving the production of superoxide ions by the pigment and further attack of the radicals on the phenol groups of the anthocyanine could account for the observed experimental evidences. These results should be carefully considered in a practical sense when rose hip oil is stored or employed under daylight conditions. Exposure to visible radiation may cause irreversible transformations in rose hip oil.     

The Role of LGR4 (GPR48) in Normal and Cancer Processes

Leucine-rich repeats containing G protein-coupled receptor 4 (LGR4) is a receptor that belongs to the superfamily of G protein-coupled receptors that can be activated by R-spondins (RSPOs), Norrin, circLGR4, and the ligand of the receptor activator of nuclear factor kappa-B (RANKL) ligands to regulate signaling pathways in normal and pathological processes. LGR4 is widely expressed in different tissues where it has multiple functions such as tissue development and maintenance. LGR4 mainly acts through the Wnt/β-catenin pathway to regulate proliferation, survival, and differentiation. In cancer, LGR4 participates in tumor progression, invasion, and metastasis. Furthermore, recent evidence reveals that LGR4 is essential for the regulation of the cancer stem cell population by controlling self-renewal and regulating stem cell properties. This review summarizes the function of LGR4 and its ligands in normal and malignant processes.     

Rancidity development during the chilled storage of farmed Coho salmon (Oncorhynchus kisutch)

Coho salmon (Oncorhynchus kisutch) is a fatty fish species whose farming production has greatly increased in recent years. Lipid damage produced during Coho salmon chilled storage was studied for up to 24 d. Lipid hydrolysis (free fatty acids, FFA) and oxidation (conjugated dienes; peroxide value, PV; thiobarbituric acid index, TBA‐i; fluorescent compounds formation, FR; browning development) were determined and compared to lipid composition (polyene index, PI; astaxanthin, AX) changes and sensory assessment (rancid odour development) results. Most lipid damage indices developed slowly during storage; thus, values obtained for FFA, PV, TBA‐i and FR were in all cases under 1.5 g/100 g, 4.0 meq oxygen/kg lipid, 0.40 mg malondialdehyde/kg muscle and 0.40, respectively. Odour assessment showed a significant (p <0.05) rancidity development at day 10, when compared to starting fish material; then, non‐acceptable values were obtained at days 19 and 24. The PI analysis showed not many differences during the storage time, with the lowest mean value at day 19. AX analysis indicated a relatively high content in the white muscle, which was maintained till the end of the experiment. A low oxidation development is concluded for Coho salmon lipids when compared to other fatty fish species under the same chilling conditions. AX was found to contribute to the oxidation stability of Coho salmon lipids, due to its free radical scavenger properties.     

Crisis response strategies in Finland and Spain

Crises are common in today's unpredictable environment rising the importance of crisis response strategies. Two societies in Europe are chosen: Finland, where trust in society is generalized to institutions, and Spain, where trust in society remains on the level of individuals. Using the results of the European Communication Monitor 2013 survey on crisis communication, we explored three hypotheses: “certain types of crises are often met with certain types of strategies,” “cultural context makes some crises more likely than others,” and “cultural context makes some response strategies more likely than others.” We find evidence for all three hypotheses and discuss our findings about perceived differences in the two societies. The results show that the cultural context plays an important role for crisis communication.     

Effect of a cold‐active pectinolytic system on colour development of Malbec red wines elaborated at low temperature

The effect of a new cold‐active pectinolytic system on colour of Malbec wines was studied under the following winemaking conditions: (i) fermentation at low temperature (20 °C) and (ii) prefermentative cold maceration (PCM) (5 °C–7 days) followed by traditional fermentation (28 °C). The pectinolytic system was mainly composed of polymethylgalacturonase and pectin lyase activities, detected under similar conditions to those in winemaking (pH 3.6–20 °C). The results show that the enzyme system significantly accelerated colour extraction by reducing the maceration time necessary for vinification at low temperature and shortening the PCM stage. Enzyme‐treated wines exhibited better chromatic parameters than their controls at devatting and after 6 months of storage. The cold‐active enzyme compensated the decrease in colour extraction due to the low maceration temperature, achieving high‐quality wines with chromatic characteristics similar to those of traditional wines.     

Synthesis of PID controllers for a class of time delay systems

In this paper we use the Hermite-Biehler theorem to establish results on the design of proportional plus integral plus derivative (PID) controllers for a class of time delay systems. Using the property of interlacing at high frequencies of the class of systems considered and linear programming we obtain the set of all stabilizing PID controllers. As far as we know, previous results on the synthesis of PID controllers rely on the solution of transcendental equations. This paper also extends previous results on the synthesis of proportional controllers for a class of delay systems of retarded type to a larger class of delay systems.     

Functional and nutritional value of the Chilean seaweeds Codium fragile,Gracilaria chilensis and Macrocystis pyrifera

The nutritional composition of the edible seaweeds Codium fragile, Gracilaria chilensis and Macrocystis pyrifera was determined, including amino acid and fatty acid contents along with tocol and carotenoid contents. The results indicated that the three algae C. fragile, G. chilensis and M. pyrifera showed a high range of protein contents (13.7–10.8%), amino acid contents (1879.6–1417.7 mg/100 g dry algae) and a low content in lipids (0.7–15.0%). The most abundant fatty acids in C. fragile and G. chilensis were linolenic, oleic, and linoleic acid. δ‐Tocopherol and α‐tocopherol (677.8 and 453.5 µg/g lipid, respectively) were found in C. fragile, while in G. chilensis and M. pyrifera, γ‐tocotrienol and α‐tocopherol (263.5 and 1327.7 µg/g lipid, respectively) were found. In addition, in C. fragile and G. chilensis β‐carotene was the principal carotenoid found (197.9 and 113.7 µg/g dry algae, respectively) compared with M. pyrifera (17.4 µg/g dry algae). The composition of macronutrients (minerals, carbohydrate‐type dietetic fiber, proteins; low in lipids) and micronutrients (essential amino acids, PUFA of balanced n‐6/n‐3 proportion, β‐carotene and α‐tocopherol as source of vitamins A and E), corroborate the nutritional and biological potential of the studied algae, which constitute useful raw materials for the development of diets or ingredients for human and animal nutrition.     

Knowledge-based process planning for electronic assembly

The conventional method of electronic assembly planning is subjective and depends greatly on the skills, memory, knowledge and experience of the planners. This gives rise to inconsistencies which in turn create problems in the manufacturing environment. Since the computer logics are systematic and consistent, it is conceivable that an automatic electronic assembly planning system would have advances over manual systems. Due to the frequent advances in technology the electronic industry is a very dynamic one. This affects the effectiveness of the current process plans and the applicability of the system that generates them. In this paper important issues which address the automatic generation of process plans will be presented. The framework of a system which takes into account the dynamic nature of the environment will be introduced as an example.The original version of this paper was presented at the 2nd International Symposium on Robotics and Manufacturing (ISRAM), Albuquerque, New Mexico, 16–18 November 1988. The published proceedings of this meeting may be ordered from: CAD Laboratory for Systems/Robotics, EECE Dept., UNM, Albuquerque, NM 87131, U.S.A.     

Optical characterization of diamond-like carbon thin films non-uniform in thickness using spectroscopic reflectometry

Two formulae expressing the reflectance of non-uniform thin films are presented. The first of them corresponds to the arbitrary shapes of this non-uniformity and illuminated light spot under the assumption that this non-uniformity is sufficiently slight. The second formula corresponds to the wedge-shaped non-uniformity of arbitrary magnitude within the circular light spot. It is shown that both the formulae are suitable for the successful optical characterization of non-uniform diamond-like carbon (DLC) thin films in contrast to the usually used reflectance formula for the uniform thin films. Within the optical characterization the values of the standard deviation of non-uniformity is determined together with the correct values of the mean thickness and material parameters corresponding to the dispersion model based on the parametrization of the density of electronic states. On the basis of the material parameters determined the correct spectral dependences of the optical constants of the DLC films studied are calculated.