Differential reactivities of lysines in calmodulin complexed to phosphatase

Calmodulin and calmodulin complexed with calcineurin phosphatase were trace labeled with [3H]acetic anhydride and the incorporation of [3H]acetate into each epsilon-amino lysine of calmodulin was measured. The relative reactivities of calmodulin lysines were higher in the presence of Ca2+ than in the presence of EGTA, and the order was: Lys-75 greater than Lys-94 greater than Lys-148 greater than or equal to Lys-77 greater than Lys-13 greater than or equal to Lys-21 greater than Lys-30. The changes in relative reactivity implied a change in conformation. When calmodulin was complexed with the phosphatase, Lys-21, Lys-77, and Lys-148 were most protected, implying that these residues are at or near the interaction sites or are conformationally perturbed by the interaction. Lys-30 and Lys-75 were slightly protected, lysine 13 showed no change, while lysine 94 significantly increased in reactivity. Comparison with results obtained from myosin light chain kinase using a similar technique (Jackson, A. E., Carraway, K. L., III, Puett, D., and Brew, K. (1986) J. Biol. Chem. 261, 12226-12232) reveals that calmodulin may interact with each of the two enzymes similarly at or near Lys-21, Lys-75, and Lys-148; one difference with phosphatase is that complex formation also involved Lys-77. These findings suggest that calmodulin interacts differently with its target enzymes.     

Variation of Chemical Constituents and Antiradical Capacity of Nine Ferulago angulata Populations from Iran

The present study was designed to assess the influence of geographical factors on essential oil (EO) composition, along with antiradical potential and phytochemical contents of Ferulago angulata (Schltdl .) Boiss (Apiaceae) extracts for the first time. The aerial parts were hydrodistilled by Clevenger apparatus and subjected to gas chromatography coupled with flame ionization detector (GC/FID) and mass spectroscopy (GC/MS). The EO yields were significantly different from populations ‘Mongar’ (south‐slope, 3000 m) with 1.34±0.06 % and ‘Male‐Amiri’ (north slope, 2600 m) with 0.18±0.05 % of total oil. Thirty‐nine compounds were identified from the EOs of nine populations. α‐Pinene was the predominant component ranging from 20.84 to 49.06 % in ‘Gandomkar’ (north‐slope, 2500 m) and ‘Mongar’ (3000 m), respectively. The methanolic extract of ‘Mongar’ (north‐slope at 2500 m) possessed the highest total phenolic contents. Also, this population logically exhibited potent antiradical activity using both 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays with EC₅₀ of 42.07±4.12 μg/mL and 8.34±0.21 mmol Trolox® equivalents/g, respectively. Due to its moderate free‐radical scavenging potential and high α‐pinene content, the population ‘Mongar’ might be considered as a perspective raw material in food and phytopharmaceutical industries.     

Designing a Novel Multi-epitope Peptide Vaccine Against Pathogenic Shigella spp. Based Immunoinformatics Approaches

International Journal of Peptide Research and Therapeutics - Shigella spp. causes severe diarrhea and dysenteric disease, which known as shigellosis. Until now, no licensed vaccine is available for...     

A novel copper (II) complex activated both extrinsic and intrinsic apoptotic pathways in liver cancerous cells

Recent advances have put fundamental focus on the application of copper (II) (Cu [II]) complexes as agents for fighting against cancer. To determine whether [Cu(L)(2imi)] complex as a novel Cu complex can induce apoptosis in HepG2 as cancerous cells and L929 as normal cells via extrinsic or intrinsic apoptotic pathways, both cell lines were treated for 24 and 48 hours at IC₅₀ concentrations of [Cu(L)(2imi)] complex. Then, the expression of some apoptosis-related genes including p53, caspase-8, bcl-2, and bax were assayed by real-time polymerase chain reaction. The [Cu(L)(2imi)] complex seems to inhibit the expression of bcl-2 in complex-treated HepG2 cancerous cells following the 24- and 48-hour treatment. The complex upregulated the p53, bax, and caspase-8 genes, therefore treatment of HepG2 cancerous cells with [Cu(L)(2imi)] complex induces programmed cell death via the upregulation of relative bax/bcl-2 ratio. Finally, this copper complex triggered apoptosis in HepG2 cells via both intrinsic and extrinsic pathway, whereas treatment of normal L929 cells with this complex induce apoptosis only via intrinsic pathway with the upregulation of relative bax/bcl-2 ratio and does not affect the expression level of caspase-8 gene and does not trigger the extrinsic pathway. Finally, these results obtained from present study confirm the role of a novel Cu complex on the induction of apoptosis process in HepG2 and L929 cells by overexpression of bax, inhibition of bcl-2 and increase of the relative bax/bcl-2 ratio. These results support that the [Cu(L)(2imi)] complex is able to induce apoptosis in cancerous cells, therefore, it has a potential for development as a novel anticancer drug.     

MSLN (Mesothelin), ANTXR1 (TEM8), and MUC3A are the potent antigenic targets for CAR T cell therapy of gastric adenocarcinoma

Gastric adenocarcinoma is usually diagnosed in late stages, necessitating the use of different therapeutic modalities. Currently, antibody-based therapies have also been approved through with limited clinical efficacy. Reinforcing antibody-based immunotherapy by using chimeric antigen receptor (CAR) T cells may enhance the approach. However, the cells can cause severe on-target and off-tumor toxicities owing to their higher sensitivity to low-level antigen expressions. To address the need for safe and reliable targets, we made a bioinformatics pipeline by which we screened overexpressed genes in the disease for off-tumor sites in many normal tissues. Our inspection showed that MSLN (Mesothelin), ANTXR1 (TEM8), and MUC3A are the probable targets of CAR T cell therapy in gastric adenocarcinoma. The proposed antigenic targets might respond to the need to simultaneously target multiple antigens in a tumor matrix to prevent resistance.     

Distinct neurocomputational mechanisms support informational and socially normative conformity

A change of mind in response to social influence could be driven by informational conformity to increase accuracy, or by normative conformity to comply with social norms such as reciprocity. Disentangling the behavioural, cognitive, and neurobiological underpinnings of informational and normative conformity have proven elusive. Here, participants underwent fMRI while performing a perceptual task that involved both advice-taking and advice-giving to human and computer partners. The concurrent inclusion of 2 different social roles and 2 different social partners revealed distinct behavioural and neural markers for informational and normative conformity. Dorsal anterior cingulate cortex (dACC) BOLD response tracked informational conformity towards both human and computer but tracked normative conformity only when interacting with humans. A network of brain areas (dorsomedial prefrontal cortex (dmPFC) and temporoparietal junction (TPJ)) that tracked normative conformity increased their functional coupling with the dACC when interacting with humans. These findings enable differentiating the neural mechanisms by which different types of conformity shape social changes of mind.

When we change our mind in response to other people’s opinion, we may be motivated to be correct or to have a good relationship with others. This fMRI study disentangles the neurobiological underpinnings of these two different motives in the human brain, and shows that the second motive is absent when we interact with computers.     

Prevalence of zinc deficiency in junior high school students of Tehran City

Zinc deficiency is a health problem in many communities especially among adolescents because of pubertal growth sprout. This investigation was carried out to determine the epidemiology of zinc deficiency in junior high school students in Tehran City in 1997. This cross-sectional study was performed on 881 students (452 males and 429 females) with the mean age of 13.2±1.0 yr, who were selected by multistage random sampling method. Plasma, erythrocyte, and hair zinc levels were assayed by flame atomic absorption spectrophotometry. Anthropometric and demographic characteristics were measured and recorded on a questionnaire. Dietary intakes were evaluated by a 24-h recall method. Zinc deficiency was defined as having at least two indices from indices of erythrocyte, plasma, and hair zinc below 10 μg/mL, 100 μg/dL, and 125 μg/g of hair, respectively. The results showed that zinc deficiency prevalence was 31.1% (confidence interval: 28–34.4%). Zinc deficiency was 65%, 49%, and 1.3% based on plasma, erythrocyte, and hair zinc levels, respectively. The mean ± SD for plasma, erythrocyte, and hair zinc concentration, height-for-age, as well as weight-for-age Z scores were 95.2±17.7 μg/dL, 10.3±2.3 μg/mL, 239.4±54.4 μg/g, −0.40±0.92, and 0.12±0.91, respectively. As for dietary intake compared with the RDA, 50% of the subjects consumed less than 50% of their requirement for zinc RDA based on a 24-h dietary recall. Zinc intake in subjects was 7.5±3.7 μg, that in boys was higher than in girls. Correlation coefficients between zinc status indices were very weak. There was neither a linear nor nonlinear relationship between biochemical parameters and nutritional zinc intake. It is concluded that almost one-third to one-half of the subjects would be considered zinc deficient.     

Hi-Plex targeted sequencing is effective using DNA derived from archival dried blood spots

Many genetic epidemiology resources have collected dried blood spots (predominantly as Guthrie Cards) as an economical and efficient means of archiving sources of DNA, conferring great value to genetic screening methods that are compatible with this medium. We applied Hi-Plex to screen the breast cancer predisposition gene PALB2 in 93 Guthrie Card-derived DNA specimens previously characterized for PALB2 genetic variants via DNA derived from lymphoblastoid cell lines, whole blood, and buffy coat. Of the 93 archival Guthrie Card-derived DNAs, 92 (99%) were processed successfully and sequenced using approximately half of a MiSeq run. From these 92 DNAs, all 59 known variants were detected and no false-positive variant calls were yielded. Fully 98.13% of amplicons (5417/5520) were represented within 15-fold of the median coverage (2786 reads), and 99.98% of amplicons (5519/5520) were represented at a depth of 10 read-pairs or greater. With Hi-Plex, we show for the first time that a High-Plex amplicon-based massively parallel sequencing (MPS) system can be applied effectively to DNA prepared from dried blood spot archival specimens and, as such, can dramatically increase the scopes of both method and resource.