Biolmpacts
Volume:11 Issue: 4, Jul 2021

Further development of magnetic-based detection techniques could be of significant use in increasing the sensitivity of detection and quantification of hepatitis B virus (HBV) infection. The present work addresses the fabrication and characterization of a new bio-nano composite based on the immobilization of goat anti-HBsAg antibody on modified core-shell magnetic nanoparticles (NPs) by (3-aminopropyl) triethoxysilane (APTES), named Fe3 O4 @SiO2 /NH2 , and magnetic NPs modified by chitosan (Fe3 O4 @CS).

At the first step, Fe3 O4 was modified with the silica and APTES (Fe3 O4 @SiO2 /NH2 ) and chitosan (Fe3 O4 @CS) separately. The goat anti-HBsAg antibody was activated by two different protocols: Sodium periodate and EDC-NHS. Then the resulted composites were conjugated with activated goat anti-HBsAg IgG. An external magnet collected Bio-super magnetic NPs (BSMNPs) and the remained solution was analyzed by the Bradford method to check the amount of attached antibody to the surface of BSMNPs.

The findings indicated that activation of antibodies by sodium periodate method 15-17 µg antibody immobilized on 1 mg of super magnetic nanoparticles (SMNPs). However, in the EDC-NHS method, 8-10 µg of antibody was conjugated with 1 mg of SMNPs. The resulting bio-magnetic NPs were applied for interaction with the HBsAg target using enzyme-linked immunosorbent assay (ELISA). About 1 µg antigen attached to 1 mg SMNPs, which demonstrated that the fabricated materials are applicable in the detection scope of HBsAg.

In the present study, we developed new antibody-conjugated magnetic NPs for the detection of HBsAg using an efficient conjugation strategy. The results demonstrated that the binding capacity of Fe3 O4 @SiO2 /NH2 was comparable with commercially available products. Our designed method for conjugating anti-HBsAg antibody to a magnetic nanoparticle opens the way to produce a high capacity of magnetic NPs.

Nowadays, probiotic bacteria have been considered as a factor in the prevention and treatment of cancer, especially by induction of apoptosis. This study aimed to evaluate the cytotoxic, anti-proliferative, and apoptotic effects of the supernatant of probiotic Lactobacillus rhamnosus on HT-29 cell line.

Molecular identification of probiotic L. rhamnosus was carried out using specific primers of 16S rRNA gene and sequencing. HT-29 cells were treated with different concentrations of bacterial supernatants at 24, 48, and 72 hours. MTT assay, Annexin V-FITC, real-time PCR, cell cycle analysis, and DAPI staining tests were conducted to evaluate the induction of apoptosis. The level of cyclin D1 protein was measured by immunocytochemistry method.

The supernatant of L. rhamnosus inhibited the growth of HT-29 cancer cells in a dose- and time-dependent manner. The results of flow cytometry confirmed apoptotic cell death. Probiotic bacterial supernatant caused up-regulation of pro-apoptotic genes including caspase-3, caspase-9, and Bax. In addition, they resulted in down-regulation of Bcl2 and a decrease in expression levels of cyclin D1, cyclin E, and ERBB2 genes. Cancer cells were arrested in the G0/G1 phase of the cell cycle. The results of immunocytochemistry showed significant down-regulation of cyclin D1 protein during the 48 hours treatment with bacterial supernatant compared to the untreated cells.

The supernatant of probiotic L. rhamnosus has a great potential to inhibit the proliferation of HT-29 cells and the induction of apoptosis. L. rhamnosus might be used as a biological anti-cancer factor in the prevention and treatment of colon cancer.

Colorectal cancer (CRC) is one of the most lethal human malignancies with a global alarming rate of incidence. The development of resistance against common chemotherapeutics such as 5-fluorouracil (5- FU) remains a big burden for CRC therapy. Therefore, we investigated the effects of melatonin on the increasing 5-FU- mediated apoptosis and its underlying mechanism in SW-480 CRC cell line.

The effects of melatonin and 5- FU, alone or in combination, on cell proliferation were evaluated using an MTT assay. Further, Annexin-V Flow cytometry was used for determining the effects of melatonin and 5-FU on the apoptosis of SW-480 cell lines. The expression levels of Bax, Bcl-2, pro-caspase-3/activated caspase 3, X-linked inhibitor of apoptosis proteins (XIAP), and survivin were measured after 48hours incubation with drugs. Cellular levels of reactive oxygen species (ROS), catalase, superoxide dismutase and glutathione peroxidase were also evaluated.

Melatonin and 5-FU significantly decreased the cell proliferation of SW-480 cells. Combination of 5-FU with melatonin significantly decreased the IC50 value of 5-FU from 100 μM to 50 μM. Moreover, combination therapy increased intracellular levels of ROS and suppressed antioxidant enzymatic activities (P<0.05). Treatment with either melatonin or 5-FU resulted in the induction of apoptosis in comparison to control (P>0.05). XIAP and survivin expression levels potently decreased after combination treatment with melatonin and 5-FU (P<0.05).

We demonstrated that melatonin exerts a reversing effect on the resistance to apoptosis by targeting oxidative stress, XIAP and survivin in CRC cells. Therefore, more studies need for better understanding of underlying mechanisms for beneficial effects of combination of melatonin and 5-FU.

A new microfluidic-based method with electrochemical detection was developed for the simultaneous quantification of acetaminophen (AP) and phenylephrine (PHE) pharmaceuticals in the human blood and pharmaceuticals (e.g. tablet and drop).

The separation was achieved on a SU8/glass microchip with a 100 µm Pt working electrode that was positioned out of the channel and 2-(N-morpholino) ethanesulfonic acid was used as a running buffer (pH 7, 10 mM). Home designed modulated high voltage power supply and dual time switcher was used for controlling the injection and separation of the analytes in the unpinched injection mode.

The injection was carried out using +750 V for 7 seconds, and the separation and detection voltages were set at +1000 V and +0.9 V, respectively. Critical parameters such as detection potential, buffer concentration, injection, and separation voltage were studied in terms of their effects on the resolution, peak height, and migration times. For each analyte, the correlation coefficients were over 0.99 (n=6). The developed microchip was able to detect AP and phenylephrine simultaneously with the limit of detection of 7.9 and 5.2 (µg/mL) respectively for PHE and AP and excellent linear range of 10-200 (µg/mL). The recovery of the drugs ranged from 96% to 103%, while the repeatability of the method through inter- and intra-day was lower than 7%.

The developed method offers several advantages, including easy sample pretreatment process, simplicity, very fast analysis compared to other typical chromatographic methods. Thus, the proposed microfluidic-based method is proposed to be used as a time- and cost-effective monitoring method for the analysis of AP and PHE.

Similarity analysis of protein structure is considered as a fundamental step to give insight into the relationships between proteins. The primary step in structural alignment is looking for the optimal correspondence between residues of two structures to optimize the scoring function. An exhaustive search for finding such a correspondence between two structures is intractable.

In this paper, a hybrid method is proposed, namely GADP-align, for pairwise protein structure alignment. The proposed method looks for an optimal alignment using a hybrid method based on a genetic algorithm and an iterative dynamic programming technique. To this end, the method first creates an initial map of correspondence between secondary structure elements (SSEs) of two proteins. Then, a genetic algorithm combined with an iterative dynamic programming algorithm is employed to optimize the alignment.

The GADP-align algorithm was employed to align 10 ‘difficult to align’ protein pairs in order to evaluate its performance. The experimental study shows that the proposed hybrid method produces highly accurate alignments in comparison with the methods using exactly the dynamic programming technique. Furthermore, the proposed method prevents the local optimal traps caused by the unsuitable initial guess of the corresponding residues.

The findings of this paper demonstrate that employing the genetic algorithm along with the dynamic programming technique yields highly accurate alignments between a protein pair by exploring the global alignment and avoiding trapping in local alignments.

Eradication of Pseudomonas aeruginosa has become increasingly difficult due to its remarkable capacity to resist antibiotics. Bacteriophages have been suggested as an alternative treatment for bacterial infections.

In-situ gel-forming eye drop containing phage against P. aeruginosa keratoconjunctivitis was prepared. The Cystoviridae phage was formulated as in-situ gel-forming formulation which is a solution formulation but turns into gel when it contacts the eye. Therapeutic effectiveness of the in-situ gel forming formulation was evaluated by histological examination on day 12 post-infection.

The viscosity of selected formulation increased when it was instilled into the eye. The histological results showed edema, abscesses, and destruction of the stromal structure of cornea in groups where no in-situ gel-forming formulation was used. In the group where in-situ gel forming formulation was used, re-epithelialization and normal corneal structure were observed.

In-situ gel-forming ophthalmic formulation containing phage can be effective in the treatment of P. aeruginosa keratoconjunctivitis.

Scientific data suggest that early exposure to endocrinedisrupting chemicals (EDCs) affect -repro, -neuro, -metabolic systems, to which are added other notions such as mixtures, window and duration of exposure, trans-generational effects, and epigenetic mechanisms.

In the present narrative review, we studied the relationship between exposure to EDCs with the appearance and development of obesity.

Exposure to EDCs like Bisphenol A during the early stages of development has been shown to lead to weight gain and obesity. EDCs can interfere with endocrine signaling, affect adipocytes differentiation and endocrine function and disrupt metabolic processes, especially if exposure occurs at very low doses, in the mixture, during early development stages for several generations.

Exposure to EDCs is positively associated with obesity development. Moreover, the use of integrative approaches which mimicking environmental conditions are necessary and recommended to evaluate EDCs' effects in future studies.