Identifying specific matrix metalloproteinase-2-inhibiting peptides through phage display-based subtractive screening

Gelatinases A and B, which are members of the matrix metalloproteinase (MMP) family, play essential roles in cancer development and metastasis, as they can break down basal membranes. Therefore, the determination and inhibition of gelatinases is essential for cancer treatment. Peptides that can specifically block each gelatinase may, therefore, be useful for cancer treatment. In this study, subtractive panning was carried out using a 12-mer peptide library to identify peptides that block gelatinase A activity (MMP-2), which is a key pharmacological target. Using this method, 17 unique peptide sequences were determined. MMP-2 inhibition by these peptides was evaluated through zymogram analyses, which revealed that four peptides inhibited MMP-2 activity by at least 65%. These four peptides were synthesized and used for in vitro wound healing using human umbilical vein endothelial cells, and two peptides, AOMP12 and AOMP29, were found to inhibit wound healing by 40%. These peptides are, thus, potential candidates for MMP-2 inhibition for cancer treatment. Furthermore, our findings suggest that our substractive biopanning screening method is a suitable strategy for identifying peptides that selectively inhibit MMP-2.     

RhoGAP18B Isoforms Act on Distinct Rho-Family GTPases and Regulate Behavioral Responses to Alcohol via Cofilin

Responses to the effects of ethanol are highly conserved across organisms, with reduced responses to the sedating effects of ethanol being predictive of increased risk for human alcohol dependence. Previously, we described that regulators of actin dynamics, such as the Rho-family GTPases Rac1, Rho1, and Cdc42, alter Drosophila’s sensitivity to ethanol-induced sedation. The GTPase activating protein RhoGAP18B also affects sensitivity to ethanol. To better understand how different RhoGAP18B isoforms affect ethanol sedation, we examined them for their effects on cell shape, GTP-loading of Rho-family GTPase, activation of the actin-severing cofilin, and actin filamentation. Our results suggest that the RhoGAP18B-PA isoform acts on Cdc42, while PC and PD act via Rac1 and Rho1 to activate cofilin. In vivo, a loss-of-function mutation in the cofilin-encoding gene twinstar leads to reduced ethanol-sensitivity and acts in concert with RhoGAP18B. Different RhoGAP18B isoforms, therefore, act on distinct subsets of Rho-family GTPases to modulate cofilin activity, actin dynamics, and ethanol-induced behaviors.     

Toxic Metals in Breast Milk Samples from Ankara, Turkey: Assessment of Lead, Cadmium, Nickel, and Arsenic Levels

Toxic metals are one of the significant groups of chemical contaminants that humans are exposed to by oral, inhalation, and dermal routes. Exposure to these chemicals begins with intrauterine life and continues during lactation period at the first years of life. Breastfeeding has a much more special place than other nutrition options for infants. However, when possibility of contaminant transfer by breast milk is considered, its safety and quality is essential. Regarding infant and mother health and limited number of information on this field in Turkey, measuring contamination levels in breast milk is important. Therefore, in the present study, lead (Pb), cadmium (Cd), nickel (Ni), and arsenic (As) levels were measured by atomic absorption spectrometry in 64 breast milk samples obtained from mothers from Ankara, Turkey. Pb and Ni levels in breast milk samples were found to be 391.45?±?269.01?μg/l and 43.94?±?33.82?μg/l (mean ± SD), respectively. Cd was found only in one of 64 samples, and the level was 4.62?μg/l. As level was below the limit of quantification (LOQ, 7.6?μg/l) in all samples. These findings will accurately direct strategies and solutions of protection against contaminants in order to reduce their levels in biological fluids.     

The effect of PlantformTM bioreactor on micropropagation of Quercus robur in comparison to a conventional in vitro culture system on gelled medium,and assessment of the microenvironment influence on leaf structure

Quercus robur L. was micropropagated by axillary bud proliferation testing two different shoot culture systems: (i) on gelled medium in Microbox (plastic vessel with a strip for ventilation) and (ii) in liquid culture in Plantform^TM bioreactor (a temporary immersion system). Two different conditions of temporary immersion were assessed: 12 min/8 h (Plantform 1) and 8 min/16 h (Plantform 2). The effect of the two culture systems was evaluated also during subsequent rooting phase, carried out on gelled medium. Finally, the influence of the different culture conditions on leaf structure was considered, taking also into consideration the micromorphological characters of young leaves from in-field-grown oaks. Nodal segments, excised from established in vitro shoots and cultured on modified Woody Plant Medium, showed a higher Relative Growth Rate in Plantform than in Microbox, but culture conditions provided in Plantform 1 favored shoot and leaf hyperhydricity. Shoots cultured in Microbox or Plantform 2 presented the same percentage of rooting after their transfer on gelled rooting medium. Leaves developed in the two different microenvironments had large stomata with elliptical shape, which indicates good functionality, and formed hairs, and epicuticolar waxes. These leaf features are considered to provide a good adaptability to ex vitro conditions.     

A novel sorting sequence in the beta2-adrenergic receptor switches recycling from default to the Hrs-dependent mechanism

Plasma membrane recycling of G protein-coupled receptors can occur by at least two distinct mechanisms as follows: a "default" mechanism that occurs nonselectively, and a specifically sorted mechanism that requires the endosome-associated protein Hrs. In this study we have defined a sequence in the beta2-adrenergic receptor cytoplasmic tail that confers Hrs dependence on receptor recycling. This sequence resembles acidic dileucine class motifs found in other membrane proteins but is structurally and functionally distinct from previously identified sorting sequences. Mutation of the novel sorting sequence rendered plasma membrane recycling independent of Hrs and independent of a distal PDZ ligand required for Hrs-dependent recycling. We propose that the novel sorting sequence functions to "switch" endocytic trafficking between mechanistically distinct recycling modes, thereby explaining failure of the wild type beta2-adrenergic receptor to recycle efficiently by default.     

An Analysis of the Proportion of the U.S. Population that Ingests Soil or Other Non-Food Substances

Reliable quantitative data are lacking that document the prevalence of ingestion of soil and other “non-food” substances among U.S. children and adults. This article explores the proportion of the U.S. population that ingests substances such as soil, clay, starch, paint, or plaster. We compiled data from the National Health and Nutrition Examination Survey (NHANES) collected from years 1971–1975 (NHANES I) and 1976–1980 (NHANES II) because these particular surveys asked participants specific questions about non-food ingestion practices. We examined the prevalence of the behavior across multiple demographic variables, such as age, gender, education, and income level. Approximately 1% (NHANES II) to 2.5% (NHANES I) of the U.S. population ingests some type of non-food substance. The most notable variation across the demographic subgroups studied was the difference in estimated prevalence among young children (1 to <3 years) compared to older children and adults. Estimated prevalence was also higher among blacks compared to whites and within lower compared to higher socioeconomic groups. This analysis helps fill data gaps on the relative pattern of non-food ingestion practices on a national scale. This information provides perspective for risk assessors when evaluating exposure variables and for risk managers when weighing risk management alternatives.