Journal of Applied Biotechnology Reports
Volume:10 Issue: 2, Spring 2023

Recent metagenomic evidence broadly supports the association and causality of gut dysbiosis with the development of colorectal cancer (CRC). Probiotics and prebiotics have been proposed as new preventive and therapeutic adjuncts for CRC management. While promoting the growth of ingested probiotics, an ideal prebiotic fibre should reach the colon intact to be fermented by a distinct group of beneficial commensal bacteria. Therefore, the selection of probiotic bacteria, the growth-promoting prebiotic fibre, and a combination of both probiotics and prebiotics (known as synbiotics) are critical in preventing carcinogenesis. Despite limited clinically measurable data, findings from preclinical animal studies have recognized the functional role of synbiotics of varying genera, strains, and doses in nourishing beneficial human gut microbiome to evade cancer. Nevertheless, translating such heterogeneous-based data from different study settings and measured outcomes into evidence-based recommendations is a very challenging task. The emerging concept is that an ideal synbiotic combination may effectively modify the intestinal microflora composition which helps imprint the immune systems for lasting chemoprevention effect. The present article reviews the roles of gut microflora in colorectal carcinogenesis followed by discussing the updated evidence of prebiotic chemoprotective effects in modulating gut microflora and host immunity against CRC. Besides, it also summarized the limitations of the clinical use of probiotics and prebiotics to achieve synergism in formulating ideal synbiotics.

Microbiota is an aggregate of microorganisms that live in mammals including humans. These microorganisms, which include bacteria, viruses and fungi, reside in large numbers in the human intestine. Microbial metabolites resulting from microbiota play an important role in various types of cancer, including colorectal cancers, prostate, ovaries, and other types of cancers, and in various inflammatory diseases such as Inflammatory bowel disease (IBD), chronic kidney disease, cardiovascular diseases, etc. Types of microbial metabolites include reboxamycin, trimethylamine oxide (TMAO), lactacystine, and short-chain fatty acids which include butyrate, acetate, and propionate. All of these microbial metabolites play an important role in the physiological activity of the body and in inflammatory and cancerous diseases. Among various microbial metabolites, short-chain fatty acids have been studied and their role in immune cells, inflammatory diseases such as inflammatory bowel disease and cancers, is more pronounced than other microbial metabolites. In this regard, immune cells, especially those of acquired immunity, such as regulatory T-lymphocytes, play an important role in suppressing inflammation caused by inflammatory diseases and contribute to microbial metabolites in maintaining intestinal hemostasis. Microbial metabolites are effective elements in the development of gastrointestinal hemostasis and prevent unwanted inflammation after microbial infections, pathogens or any inflammatory disorders as far as possible. Microbial metabolites can help eliminate tumor cells by induction of apoptosis and specific mechanisms that will be discussed in this article. This review looks at the role of microbial metabolites in cancer and inflammatory diseases, especially IBD, and their association with the immune system.

Solenostemma argel is a Saharan plant used in traditional medicine to cure pain. The present work investigated the quantitative analysis of the composition of the essential oil of S. argel leaves (EOSA) as well as its acute toxicity and anti-nociceptive activity. The chemical characterization of EOSA was carried out by GC-MS and NMR. EOSA acute oral toxicity study was performed according to the OECD-420 method. EOSA anti-nociceptive activities were evaluated by acetic acid-induced abdominal writhing test, hot plate test, and formalin test. Twenty components were identified by GC-MS including Linanool (57.10%), terpineol (12.95%), trans-geraniol (12.65%), and nerol (4.67%). The main compound linalool was isolated by NMR. The EOSA at 250 and 400 mg/kg significantly attenuated acetic acid-induced writhing by 72.71 and 92.41%, respectively. Moreover, Ingestion of EOSA at doses of 250 and 400 mg/kg caused a significant and dose-dependent anti-nociceptive effect in both neurogenic and inflammatory phases of formalin-induced licking. EOSA impacts the pain latency in the hot plate test. The results of this study showed that EOSA has an anti-nociceptive effect on central and peripheral pain.

Bacterial food poisoning is considered a global concern in terms of economical and human health. Staphylococcus aureus produces low molecular weight extracellular toxins. These enterotoxins (SEs) are similar in structure and bioactivity. Today, there are several methods to detect SE genes. Nevertheless, culture-based and immunological methods are simple and cheaper than molecular techniques, they are time-consuming, with low sensitivity and specificity. In this field, loop-mediated isothermal amplification (LAMP) is a fast and simple method for gene amplification. The aim of this work was to optimize the LAMP reaction using the Taguchi method. For this, in order to improve and accelerate the LAMP diagnostic process, essential factors for the identification of enterotoxin, including MgSO4 concentration, time, and temperature reaction were optimized separately and using Taguchi experimental design. The results showed that 57 µg/ml of S. aureus genome as template is suitable for the replication, and in optimization of LAMP assay in separate condition the best replication rate was observed at 6 mM MgSO4, 45 min, and 65 °C. Whereas using Taguchi methods, the optimum condition was at 8 mM MgSO4, 60 min and 65 °C. The one-step-visual developed LAMP assay with the optimum conditions could be of interest for screening functions in food analytical laboratories, and a portable detection method could be used to design a suitable identification kit for S. aureus without the need for special equipment or trained personnel.

Arbutus unedo L., is an evergreen plant belonging to the Ericaceae family, an endemic species of the Mediterranean flora. The aim of this study was to characterize the polyphenolic extract of A. unedo fruits with phytochemical analysis followed by evaluation of antioxidant and anti-inflammatory activities. Antioxidant activity was determined using the scavenging activity of DPPH free radical and ferric-reducing antioxidant power assay. Then, the anti-inflammatory potential of A. unedo extracts was evaluated using Human Red Blood Cells (HRBC) membrane stabilization, and egg albumin denaturation assays. The highest total phenolic, flavonoid, and tannin content was recorded in the methanolic extract with 61.96 ± 5.33 mg GAE/g, 51.16 ± 0.57 mg QE/g, and 2.40 ± 0.14 mg CE/g, respectively. On the other hand, the best radical scavenging activity (IC50 = 0.459 ± 0.022 mg/ml) and the highest reducing power activity (EC50 = 0.471 ± 0.022 mg/ml) were exhibited by the methanolic extract A. unedo. Whereas regarding the anti-inflammatory activities, A. unedo aqueous extract exerted the highest HRBC stabilization of 70.86 ± 0.61% and Egg albumin denaturation inhibition of 70.06 ± 0.68%. Overall, the results suggest that aqueous and methanolic extracts of A. unedo fruits can be used as future ethnomedicinal antioxidants and anti-inflammatories due to their rich content of bioactive molecules.

Botulinum neurotoxin is one of the most potent toxins. This neurotoxin is a biological weapon due to its high lethality and simplicity of preparation. The receptor-binding domain of botulinum neurotoxin is a promising vaccine candidate. In this study, the immunogenicity of the C-terminal domain of botulinum neurotoxin type A binding domain was investigated.

The synthetic gene encoding 285 C-terminal amino acids of BoNT/A binding domain fused to trxA gene, was constructed. The sequence was optimized codon usage for expression in E. coli and subcloned into pET-17b expression vector. The recombinant protein was expressed using 1 mM IPTG and purified by affinity chromatography on a column of Ni-NTA. The recombinant protein was confirmed by Western blotting and was used to immunize mice. The indirect ELISA and t-test were used to assess and compare antibody titers against recombinant protein.

The codon adaptation index of the contract was altered from 0.62 to 0.90 after optimization. The minimum energy of the predicted mRNA structure was (-308.39) kcal/mol. SDS-PAGE and Western blotting confirmed the 44/6 kDa recombinant protein. Following immunization, mice elicited significant IgG antibodies in serum compared to control mice (p<0.05).

The results indicated a highly expressed and purified recombinant protein, which is able to evoke high antibody titers in mice. Future studies may develop the recombinant antigen as a potential immunogenic candidate against botulinum neurotoxin type A.

Radiotherapy is a standard and effective modality for breast cancer treatment, through induction of DNA damages notably DNA double-strand breaks which are involved in radiation-induced cell death. All radiation-induced DNA damages are subjected to various repair processes, therefore, interference in the DNA repair pathways might result in radio-resistance. Non-coding RNAs are a diverse group of functional RNA molecules that are not translated into proteins. Recent studies have shown that radiation can cause expression changes in noncoding RNAs. MCF-7 and MDA-MB-231 cell lines were grown in a DMEM culture medium supplemented with fetal bovine serum and antibiotics. At exponential growth, cells were exposed to various doses of megavoltage X-rays. 24 and 48 h after irradiation cells were harvested, RNA was extracted and cDNA was synthesized. The expression level of lncRNAs was measured using quantitative real-time PCR. This study showed that radiation could increase DANCR and TUG1 lncRNAs expression in breast cancer cell lines 24 and 48 h after receiving radiation. Also, the results suggested that after radiation, the expression of DANCR in the radioresistant cell line was higher than the radiosensitive one; in the case of TUG1, it’s unlike DANCR. The radiation increased the expression of DANCR and TUG1 lncRNAs in breast cancer cell lines because the expression of DANCR in the MDA-MB- 231 was higher than in the MCF-7. In contrast, the expression level of TUG1 in MCF-7 was higher than the MDA-MB-231. Therefore, lncRNAs, DNACR, and TUG1 might play a role in the radioresistance and radiosensitivity of breast cancer, respectively.

COVID-19 virus has caused the biggest pandemic in a decade. The acute respiratory syndrome caused by this virus can lead to the death of patients. Death is very likely in people with severe forms of lung disease. Early identification of patients with severe disease can be very effective in the prevention of death outcomes with improves triage strategies and timely medical actions. The aim of this study was the prediction of COVID-19 models for death with three different decision algorithms with analysis of hospitalized patients. In this study, in a retrospective analysis of 600 COVID-19 patients, we apply three decision tree algorithms including the C5.0, CRT, and CHAID using all related factors to the disease including demographic data, history of exposure, clinical signs, and symptoms, laboratory results, chest X-ray or computed tomography (CT) scans, underlying illness, treatment steps, and outcomes of each patient to build several models predicting the death of Covid-19 infection. The accuracy of the models was above 90%. Overall, in our retrospective analysis, age, hypertension, lung disease, O2Sat, diabetes, and body temperature, respectively are the most important factors that can affect the mortality rate of COVID-19 patients. Among them, age, and hypertension are common in our applied three models. The design of such models and apply in hospitals can help to improve disease management and decrease the mortality rate spatially in about recent pandemic.

Invertase belongs to the class of enzymes called glycosidase. The enzyme is responsible for the catalytic hydrolysis of sucrose to release monosaccharides known as invert sugars. The aim of this study was the isolation, purification, and physicochemical properties of intracellular invertase from palm wine yeast (Saccharomyces sp.) as an alternative enzyme in several industrial applications. The yeast was harvested from the palm wine through flocculation and the intracellular invertase was isolated from the yeast cell by mechanical grinding using acid washed sand. The intracellular invertase was purified using combination of ion-exchange chromatography on DEAE-trisacryl and gel filtration on Sephacryl S-300. The kinetics and other physicochemical properties of the purified enzyme were determined. The two-step purification scheme employed gave a final yield of 168% and a purification fold of 3.0. Denaturing polyacrylamide gel electrophoresis (SDS-PAGE) of the intracellular invertase gave six subunits with the molecular mass of 73.5 ± 2.1 kDa, 52.3 ± 8.1 kDa, 48.6 ± 3.0 kDa, 37.4 ± 4.8 kDa, 26.5 ± 3.6, and 24 ± 4.3 kDa, respectively, while non-denaturing PAGE revealed the presence of single entity invertase with the molecular mass of 267 kDa. Also, native molecular mass estimated on calibrated Sephacryl S-300 was 266 ± 28 kDa, revealing that the purified palm wine Saccharomyces invertase (PWSInv) is heterohexameric in nature. The Km and Vmax of the purified invertase were 30.8 ± 3.2 mM and 9672 ± 169.0 units/mg protein, respectively, leading to catalytic efficiency, kcat/Km of 1.43 × 106 M-1 s-1. The optimum temperature and pH were 60 °C and 3.0, respectively. The activation energy (Ea) of the intracellular invertase for the hydrolysis of sucrose was estimated to be 280.42 kJ/mol. The study established the presence of intracellular invertase from palm wine yeast and investigated some properties and characteristics of the purified invertase, which could be exploited for several biotechnological and industrial processes.