Biomacromolecular Journal
Volume:4 Issue: 1, Summer 2018

Lead has many applications in the industry, but does not play a vital physiological role in the human body. On the other hand, lead has shown undesirable effects on the nervous, gastrointestinal, and endocrine systems, and people who are highly exposed to this element, due to their occupations or residencies, are affected by its harmful effects. Therefore, we designed a cross-sectional study on traffic officers with high exposure level to lead in Tehran, Iran, to specify the effect of lead on their levels of thyroid hormones and blood cells. The blood lead concentration was measured using Lead Analyzer and Flame Atomic Absorption spectrometry. The levels of levels of thyroid hormones and blood cells were measured using KX-21N and ADVIA Centaur XPT (Immunoassay system) instruments, respectively. The obtained results demonstrated that there was no significant association between blood lead levels and levels of thyroid hormones, number of blood cells, and related blood cells parameters. However, there was a weak association between blood lead levels and the age of participants (r = 0.25). Further studies with higher sample size could confirm these results.

Cellulase is one of the most commercial and applicable enzymes which is produced chiefly by fungi, bacteria, protozoa and metazoa that catalyze cellulolysis, the decomposition of cellulose and of some related polysaccharides. Recently, the demand for this enzyme has been increased mostly for the various industrial purposes. It has been assumed that the milieu of earthworm’s intestine is the final destination of digestive enzymes such as cellulase. For the first time, this study is aimed to shed a light on the characteristics of cellulase activity in the coelomic fluid and body extract of the earthworm Allolobophora chlorotica. Practically, following the preparation of homogenate and coelomic body in cold-ice and enzyme activities at different conditions were investigated and based on results, the optimum pH and temperature for cellulase activity were pH 9 and 50℃, respectively for the two sources. Given the results, the earthworm contains a novel cellulolytic enzyme which is more stable at alkaline pH. A side from optimal pH and temperature, the addition of metal salts, including CaCl2, NaCl, MgCl2, KCl increase cellulase activity, although, HgCl2 and EDTA have inhibitory effects on the enzyme. Due to common properties of cellulase from coelomic and body extracts it can be concluded that coelomic cellulase contents can be secreted from digestive tracts to facilitate the digestion of the food. Furthermore, based on results, the novel cellulase is active at alkaline pH and moderately thermostable which can be used in biotechnological industries after purification.

Due to the importance of proteins in the differentiation and function of cells, investigation of variation of the protein expression levels can help for the better understanding of the differentiation processes. Cell-therapy provides a promising alternative for the treatment of type 1 diabetes. In our previous studies, we isolated and expanded human skin-derived precursors (hSKPs) and differentiate them into IPCs in vitro, through exposure to suitable differentiation factors. Cellular clusters were also able to secrete detectable amounts of insulin and C-peptide in glucose concentration dependent manner. In the present study, we applied two-dimensional electrophoresis technique to evaluate variations in protein expression in expanded SKPs and differentiated IPCs. The results showed that the protein pattern of SKPs differs with differentiated IPCs in terms of the number of points and the expression level of protein. The current study demonstrates that IPC differentiation in human SKPs is accompanied by modulation of 13 major proteins with diverse Functions: metabolic, chaperone activity, redox (antioxidant), protein degradation, transcription and signaling cascades. Overexpression of three key enzymes in the glycolytic pathway was observed. Since insulin is a hormone that is involved in glucose metabolism, it is not surprising overexpression of glycolytic enzymes found in differentiated cells. Here we show for the first time the expression of the antioxidant proteins in differentiated cells has increased. Also augmentation in ubiquitin and cyclophilin A was observed, indicating overexpression of proteins that involve in degradation of old proteins and synthesis and folding of new proteins in differentiation process.

β-lactoglobulin (β-LG) is the major protein in the whey of ruminant milk and it can be used as a carrier for anticancer drugs. In this study, the comparison interaction of two chemotherapeutic anti-cancer drugs of 5-Fluorouracil and oxali-palladium with milk carrier protein of β-LG was investigated using fluorescence spectroscopy. The analysis of fluorescence spectra showed that the addition of the 5-Fluorouracil and oxali-palladium to β-LG solution led to a significant reduction in the intrinsic fluorescence spectra of protein and then quenched it. The binding sites and thermodynamic parameters of 5-Fluorouracil and oxali-palladium on the protein were calculated by analyzing of van’t Hoff equation at different temperatures. Binding results have represented that there are 2 and 1binding sites on β-LG for binding of 5-Fluorouracil and oxali-palladium at physiological temperature, respectively. According to the results, van der Waals and hydrogen bonding have the main role of the interactions of β-LG with oxali-palladium, while electrostatic interactions have a major role in the interaction of β-LG with 5-Fluorouracil. Finally, regard to the above results, it can be concluded that the anticancer drugs of 5-Fluorouracil and oxali-palladium can bind to the carrier protein of β-LG and changed the structure of it differently.

Amylases are a family of endoamylases that randomly hydrolyze starch. This study for the first time aimed to explore α-amylase enzyme activity and the existence of amylases in coelomic fluid and body extract from the earthworm Allolobophora chlorotic. In practice, the worms were homogenized in ice-cold water and centrifuged to obtain a supernatant form earthworm body. Moreover, to prepare coelomic fluid electric shock was used and subsequently the activities of amylase in coeloimc fluid and body extract were measured at different pHs and temperatures. Based on results, the optimal pH and temperature for amylase activity in the body extract and coelomic fluid were 8 and 50°C, respectively. Additionally, thermostability and pH stability of the enzyme was measured at different temperatures and pHs. At the other extreme, results indicated that metal salts, including CuSO4, CaCl2, NaCl, MgCl2, KCl have enhanced amylase activity and, HgCl2 and EDTA have inhibited the enzyme form both sources. As a consequence, it can be concluded that novel amylases in coeloimc fluid and body extract from the earthworm Allolobophora chlorotica behave similarly and the amylase can be purified in large scale from the earthworms and exploited in various biotechnological applications.

The aim of this study was to evaluate anti bacterial and antibiofilm activity of PCL/gelatin nanofibers containing Lawsone against four pathogenic bacteria. Different concentrations of Lawsone (2-hydroxy-1, 4- naphthoquinone) (1, 3, 5 and 10%) were incorporated into polycaprolactone (PCL), gelatin (GEL) nanofibers via electrospinning technique. Presence of Lawsone in the scaffold confirmed by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Antibacterial and antibiofilm activity of Lawsone loaded scaffolds against clinically isolated Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and Proteus mirabilis were investigated by disc diffusion method. All scaffolds were able to inhibit the growth of S. aureus and MRSA. Additionally, PCL/GEL/Lawsone 10% had bactericidal efficacy against P. mirabilis. Obtained results of the biofilm formation assay revealed that PCL/GEL/Lawsone 10% inhibited the biofilm of S. aureus, MRSA and P. mirabilis by 56%, 32%, and 15%, respectively. Due to good antibacterial and antibiofilm activity of Lawsone containing scaffolds, PCL/GEL 10% can be a good candidate for application in wound dressing patches.

Endostatin, the C-terminal fragment of collagen XVIII, is known as an endogenous inhibitor of angiogenesis and is currently used as an anticancer drug. Endostatin fragments can be used as alternatives for full-length molecule, among which a peptide derived from the C-terminal fragment of protein, including residues 123-184 was shown to effectively inhibit angiogenesis and tumor growth. The aim of current study was to design a shortened peptide of the fragment 123-184. For this purpose, the presumably non-functional segments were deleted, including β-hairpin segment, comprising residues 145-163 and C-terminus of the protein, including residues 175-184. The designed 30-amino acid peptide that encompasses random coils in 123-184 (referred to as C-peptide) was synthesized and characterized. C-peptide inhibited the proliferation of the Human Umbilical Vein Endothelial Cells (HUVECs) with an IC50 value of 0.35 µM. Administration of C-peptide caused the regression of 4T1 murine mammary carcinoma tumor growth, considerable reduced tumor cell proliferation (Ki67 expression) and angiogenesis (CD31 and CD34 expression), and the induction of apoptosis (increased TUNEL staining). These results confirm that the random coils of the C-terminal domain of endostatin are implicated in its antiangiogenic and antitumor properties.