Volum 27 Nr. 3 (2022): Romanian Biotechnological Letters
Acest număr publică articole originale, recenzii de literatură, comunicări științifice scurte în domeniul biotehnologiilor (verzi, rosii, albe, albastre, bioinformatică) și al cercetării interdisciplinare în medicină, neuroștiințe, agricultură, zootehnie, ecologie și protecția mediului.
Număr complet
Articles
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Relative leaf expansion rate as an indicator of compensatory growth of defoliated vines of Prokupac (Vitis vinifera L.)
Rezumat
Reaction of the grapevine to the early defoliation is to mitigate its effects through compensatory growth by producing more lateral shoots with a greater number of leaves. In this study, we evaluated the usage of non-destructive and continuous measurements of mean and lateral leaf area on the same shoots for the purpose of monitoring the leaf area development and calculating relative leaf expansion rate - RLER during vegetation. This study has shown that the grapevine's ability to recover its leaf area after defoliation depends mainly on the time when the defoliation occurs. Early defoliated vines had time to compensate removed leaves by producing more lateral shoots with a greater number of leaves, which resulted in a larger leaf area. With the decrease in the intensity of shoot growth during vegetation, the recovery ability decreases, therefore, compensatory growth is not enough to restore the reduced leaf area. Based on the value of RLER, its defoliation is performed in the period of intensive growth of shoots, it affects the stagnation of the emergence of new shoots and leaves over several days, followed by a period of re-growth. Very slow or no growth of shoots and leaves occurred after the veraison stage.
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Canine eosinophilic bronchopneumopathy: case study
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Eosinophilic bronchopneumopathy, formerly known as PIE (Pulmonary Infiltration with Eosinophils) syndrome, is defined as an inflammatory pathology haracterized by the presence of eosinophilic infiltrate in the lungs and bronchiolary structures. (CECILE CLERCX,2007). Canine eosinophilic bronchopneumopathy has been described similarly to that present in humans, highlighting tropism for the airways and lungs, but also the multitude of factors with the potential to trigger, which is why most of them are ranked with idiopathic factor. (Sol Kim et a1,2021). Cases of eosinophilic bronchopneumopathy have been identified in most canine breeds, and according to current studies, the predisposition for large breeds under the age of 6, such as rottweilers, Siberian Husky and Alaskan Malamute, has been reported. (C. Clercx 2017). The present case is represented by a male, canine, of the Husky breed, who showed clinical respiratory signs, these were correlated with complementary examinations such as laboratory examinations, cardiological examination, radiological and endoscopic examination. Based on the haematological results, which present a marked eosinophilia, a differentiated diagnostic protocol was achieved compared to possible parasitosis and reactions of autoimmune type. Differential clinical diagnosis was made against dirofilariosis, angiostrongiliosis, lupus erythematosus, mycotic pneumopathies. PCR technique was used to determine the possible parasites present, sample collection with the help of endoscopy from the pulmonary infiltrate with cytological examination. The diagnosis of certainty was established on the basis of histopathological examination. He underwent treatment with corticosteroids, the evolution being favorable within 14 days of initialization of the treatment, with complete healing.
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Use of Stabilized rice bran for preparing Beef Burger
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This study was performed to investigate the effect of thermal processing (microwave oven, roasting by dry heat at 130°, for 10 and 20 min. and steaming by using an autoclave under atmospheric pressure for 10 and 20 min.) on the stability of rice bran and the utilization of microwave stabilized rice bran as fat replacers in preparing a low-fat beef burger. The obtained results revealed that: rice bran contained high levels of carbohydrates and moderate amounts of ether extract and crude protein. No differences were detected in the refractive index of rice bran oil among the stabilization procedures, whereas slight differences were found in the case of specific gravity. When rice bran oil was subjected to stabilized procedures, the acid value, iodine value, peroxide value, saponification value, and unsaponifiable matter were reduced. Moreover, the total saturated fatty acids content of rice bran oil increased as a result of subjecting to stabilization procedures whereas the total unsaturated fatty acid content decreased. The results show that stabilized rice bran is effective in improving the chemical and functional characteristics of a beef burger.
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Effect of Partially Substitution of Rice bran and Soybean Protein Concentrates on Chemical, Nutritional and Organoleptic Properties of Kapreeta tuna-like fish (Scombromorous Spp.) Surimi
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The objective of this research is to study the possibility of utilizing kapreeta fish (like tuna) that has
low nutritional value because of its bloody dark flesh and presence of many blood vessels and susceptibility to rapid spoilage, therefore. Kapreeta fish was used to prepare a high nutritional value of Surimi blends prepared from fish flesh supplemented with 15% level of soybean and rice bran protein concentrates to suggest its protein as a new protein source. Various compositional and functional properties were assessed to determine the effect of this substitution treatment on the final product characteristics. The obtained results revealed that: rice bran protein concentrate is considered a good source of protein (72.90% on dry matter), more than 48.10 and 35.60% for soybean protein concentrate, whereas ash content of soybean protein (5.83%) are higher than that its value of rice bran protein concentrate (4.47%). The two protein sources showed to be rich in essential amino acids and met human requirements for all the essential amino acids since they have higher values than that recommended by FAO/WHO (1991) pattern for children of 10-12 years.
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Effect of ascorbic acid and tri-sodium phosphate treatment at different packaging conditions on cold storage Bagrus bayad fish quality
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This research paper aimed at evaluating the influence of ascorbic acid (AA) and tri-sodium phosphate (TSP) under different packaging condition on improving the shelf life and qualities of fillets of Bagrus bayad cold fish at 4±1°C for 12 days. The microbiological analysis performed were the total bacterial count (TBC), proteolytic bacterial count (PBC), lipolytic bacterial count (LBC), Coliform count (CC), Staphylococcus aureus (SA), Salmonella & Shigella (SS) and yeast & mold (YM). The obtained results indicated that all noticed analysis parameters (TBC, PBC, LBC, CC, SA, SS, YM, AA and TSP).were gradually increased during storage period in different treatments. The increases of these parameters were significantly higher (p <0.05) in control sample than samples treatment with (AA) and (TSP). The obtained results also showed that there was a significant (p<0.05) extension of the shelf-life through delaying the microbial growth of the samples treated with (TSP) and (AA), respectively. In addition, the shelf-life and the storage period for these products were found to be less than 3 days for control sample and more than 12 days of samples treated with (TSP) under cold storage (4°C). Therefore, the (TSP) had a high effectiveness for extending the shelf-life and enhancing quality attributes of raw fish fillets during cold storage.
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Direct-Contact and Contact-free Treatment of a DBD-like Non-thermal Atmospheric Pressure Plasma Jet on Pseudomonas aeruginosa
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In this study, a Dielectric Barrier Discharge (DBD)-like non-thermal atmospheric pressure plasma jet (NTAPPJ) was exposed to the Pseudomonas aeruginosa planktonic bacteria. The P. aeruginosa bacteria were treated by the NTAPPJs in both Ar and He at the non-contact distance (contact-free) and the contact distance (direct-contact). The contact of the NTAPPJs with the bacteria has generated spark discharges in both filamentary mode and diffuse mode. The antibacterial efficacy of the Ar and He NTAPPJs for various treatment times (60, 120, 180, and 300 sec) were investigated by treatment on the bacteria without contact and with contact in both filamentary mode and diffuse mode. The NTAPPJs in both Ar and He caused erosion on the agar surface when treated in direct-contact with the bacteria in the filamentary mode, while the NTAPPJs in Ar in the diffuse mode produced the greatest antibacterial efficacy without damage.
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PETase from Ideonella sakaiensis: towards facile bacterial expression system
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One of the most promising PET-hydrolysing enzyme, IsPETase, originates from 'plastic-consuming' bacteria Ideonella sakaiensis and operates at moderate temperatures (30-40 °C) in aqueous environment, showing far higher hydrolysing efficiency than several cutinases. Accordingly, the development of proper recombinant expression system providing facile access to the isolated/purified IsPETase is also of high interest. In our aim to produce active recombinant IsPETase, the unsuccessful expression registered by employing several reported expression systems, directed us towards the optimization/ development of a facile/accessible recombinant expression system suitable for IsPETase production. Testing several plasmid constructs, providing different N- or C-terminal affinity tags for the expression of full-length or truncated (residues 28-290) IsPETase and various E. coli expression hosts, revealed several non-reported issues hindering the recombinant expression of IsPETase. After optimization of the construct and expression host, the use of N-terminal His-tag and Rosetta-gami B expression host provided recombinant IsPETase in high purity and titer-yield. The thermal denaturation profile and PET-hydrolysing activity of the obtained IsPETase agree with the reported data, supporting proper folding of the purified protein. The results support that the in vivo folding process of IsPETase might be differently affected among the different E. coli host strains, moreover, underline the importance of the proper selection of the cloning strategy for the successful expression of the IsPETase.
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Extraction optimization, characterization and anti- inflammatory activity of functional protein from Mangifera indica seeds kernel
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Mango seed kernel (MSK) was observed to be a likely source of bioactive protein. Box-Behnken Design (BBD) was employed to optimize the protein extraction conditions. The optimum extraction conditions for maximum protein yield (68.336%) were corresponding to extraction temperature 54.53 °C, buffer-to-sample values 41.79 mL/g and extraction time 120 min with desirability value of 0.471%. The results of electrophoresis patterns of MSKP revealed two types of proteins; low molecular weight between 6 to 98 kDa explained the presence of globulin with type (2S) and another principal protein band of 27.6 kDa explained the presence of lectins. The extracted proteins under optimized conditions showed inhibition activity of nitric oxide (NO) release and pro-inflammatory inducers like (IL-6), (IL-1r3) and (TNF-a) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. These findings revealed that MSK could be used as a promising nutraceutical of functional and health-promoting proteins.
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Characterization of exopolysaccharide produced by Ganoderma sp. TP and its immunomodulatory properties
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Mushrooms have an excellent nutritional value and are referred to as healthy food. They are a good source of carbohydrates, proteins, vitamins, and minerals while they are low in fat content. Mushrooms are well known as rich sources of nutraceutical molecules also. They produce exopolysaccharides (EPS), having high antioxidant activity. Hence, the present study aims to characterize a mushroom exopolysaccharide for its antioxidant and antitumor activity. A mushroom sample was collected from the Naysari Agricultural University, Naysari campus (20.9302° N, 72.9127° E). The sample was identified by morphological and 18S rRNA as Ganoderma sp TP (Accession No. MF614913). EPS was extracted from the sample, and total carbohydrate 258.3±1.4 gg/m1 content was measured. Chemical characterization by FTIR, TLC, HPLC, and NMR analysis revealed that arabinose, lactose, and ribose monomer were present in the structure. TGA analysis revealed 86.34 % weight loss after the second stage. Rheological analysis suggested it is dilatants in nature. Antioxidant activity measured by ABTS and DPPH showed 84.09 % and 86.45% antioxidant activity, respectively. Antitumor activity was examined using the MDA-MB-231 cell line and found 63.49% cell inhibition compared to control. Thus, EPS produced by Ganoderma sp TP is novel from other reported EPS and positively affects tumor cell growth reduction.
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In vitro elicitation supports the enrichment of 2H4MB production in callus suspension cultures of D. hamiltonii Wight & Arn
Rezumat
Influence of elicitation on in vitro production of 2-Hydroxy-4-Methoxy Benzaldehyde (2H4MB), a structural isomer of vanillin from callus suspension cultures of Decalepis hamiltonii was investigated. In vitro culture conditions were optimized to induce callus, suspension culture and biomass followed by metabolite production. Suspension cultures were established using leaf generated friable callus. Maximum content of 2H4MB production 0.079 ± 0.01 mg 100g-1 DW and biomass 197.5 ± 1.5 g1_,-1 were observed by 4th week of culturing. Elicitation was induced to suspension cultures by using mtopolin (mT), sodium nitroprusside (SNP), and pectin individually at different concentrations. mtopolin (15 itiM), pectin (15 itiM) and SNP (10 itiM) supported 0.31 mg100g-1DW, 0.27 mg100g-1DW and 0.21 mg100g-lof 2H4MB production respectively by 4th week. This data infers that the elicitation improves 2H4MB content in callus suspension cultures of D. hamiltonii.
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Assessment of the antimicrobial activity of some essential oils against multi-resistant bacterial strains
Rezumat
Volatile oils, also called essential oils, are secreted by plant-specific secretory cells and tissues. These components are a mixture of different substances, primarily mono and sesquiterpenes. Our aim was to assess the antimicrobial activity of different essential oils, i.e. lavender, cloves, eucalyptus, rosemary, thyme and oregano against multidrug resistant bacterial strains belonging to the ESKAPE group. The most effective oils proved to be the thyme and oregano which induced the occurrence of the largest diameters of the growth inhibition zones against all tested bacterial species. Oregano oil showed the best inhibitory activity against Pseudomonas aeruginosa, followed by clove, while thyme oil was the most effective against Staphylococcus aureus strains.
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Effects of prebiotics on acid and bile resistance of Bifidobacterium lactis and Lactobacillus acidophilus probiotic bacteria
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The effects of prebiotics on acid and bile resistance of probiotics were evaluated. Two commercial strains, Bifidobacterium lactis BB-12 and Lactobacillus acidophilus LA-5 had higher acid and bile resistance characteristics than the ATCC strains. B. lactis BB-12 was the most resistant to acid and bile and showed higher survival capacity. The counts of B. lactis BB-12 viable cells in pH 2 medium decreased by 2.7 log unit while the viability of other strains decreased by 4.5-7 log units. The counts of B. lactis BB-12 viable cells in the bile medium after 6 and 24 hours of incubation were determined as 6.1 and 2.8 log CFU/ml, respectively, while no viable bacteria could be determined for the other strains after 24 hours of incubation. The acid and bile resistance of probiotic bacteria remarkably varied with the type of prebiotics. Acid and bile resistance properties of L. acidophilus LA-5 and L. acidophilus ATCC 4356 were found higher when INU was present in the growth medium. Acid and bile resistance properties of B. bifidum ATCC 15969 was higher when the medium contained GOS whereas acid and bile resistance of B. lactis BB-12 was found to be higher when medium contained XOS.
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Impact of ligustrazine on chondrocyte apoptosis in knee osteoarthritis through FAK/PI3K/Akt pathway
Rezumat
Objective: To probe into the impact of ligustrazine on chondrocyte apoptosis in knee osteoarthritis (OA) and its possible mechanism.
Methods: Our team extracted the human primary chondrocytes to construct the IL-113-induced chondrocyte damage model, detected the impact of ligustrazine on chondrocyte apoptosis through TUNEL and Western blot, applied Western blot to detect the impact of ligustrazine on matrix-degrading enzymes, matrix-associated proteins, as well as FAK/PI3K/ Akt pathway, using ELISA detected the impact of ligustrazine on MCP-1, IL8, PEG2 and NO levels, applied FAK/PI3K/ Akt pathway inhibitor PF573228 to inhibit FAK/PI3K/Akt pathway to detect whether the chondroprotective impact of ligustrazine was inhibited, constructed a New Zealand rabbit osteoarthritis model, applied HE staining to evaluate the impacts of ligustrazine and ligustrazine+PF573228 on cartilage histomorphology, and applied Western blot to detect the impacts of ligustrazine and ligustrazine+PF573228 on apoptotic proteins, matrix-degrading enzymes and matrix-related proteins in cartilage tissue.
Results: Ligustrazine signally inhibited IL-113-induced chondrocyte apoptosis, reduced Bax, caspase-9, caspase-3, caspase-7, MMP3, MMP13, collagen X protein expression levels, and reduced MCP-1, IL8, PEG2, and MCP-1. NO contents, promoted collagen II and aggrecan protein expression. Additionally, ligustrazine activated the FAK/PI3K/ Akt pathway, and applying PF573228 to inhibit the FAK/PI3K/Akt pathway abolished the chondroprotective impact of ligustrazine. At the in vivo level, ligustrazine activated the FAK/PI3K/Akt pathway to inhibit Bax, caspase-9, caspase-3, caspase-7, MMP3, MMP13, collagen X protein expression levels, promoted collagen II and aggrecan protein expression, and improved cartilage histomorphology.
Conclusion: Ligustrazine exerts chondroprotective impact through FAK/PI3K/Akt pathway
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Monkeypox Virus review
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In 1958, the first case human infected by Poxivirus was detected in Democratic Republic of the Congo. This virus can produce after an incubation period of 5 to 21 a large spectrum of symptoms which can be severe, and, in many cases, medical care is necessary. The research of Monkeypox requires special laboratory conditions. Different tests can help to identify this virus in human specimens, like PCR and ELISA. Vaccinia vaccines prepared from viral variants can offer protection against monkeypox.
