Avicenna Journal of Medical Biochemistry
Volume:10 Issue: 1, Jun 2022

Camel milk has been recognized for its health benefits since ancient times and has recently been attracting increasing attention as a form of medical treatment for diverse human diseases. Studies on the health benefits of camel milk attributed its medicinal effects to nutritional status, but the molecular mechanisms of proteins involved in such effects remain unknown.

The aim of this study was to explore the anticancer properties of camel milk proteins (CMPs).

CMPs were fractionated into camel casein proteins (CCPs) and camel whey proteins (CWPs). The CWP exhibited the most potent anticancer activity against colon (HCT-116) and breast (MCF-7) cancer cells. The CWP was further fractionated into cationic and anionic proteins using HiTrap cationic (SP-XL) and anionic (QFF) exchange columns.

QFF-bound proteins (QFF-B) exhibited the strongest anticancer activities against both cancer cells. QFF-B proteins produced three peaks (P1~P3) on RP-HPLC, whereas P3 showed superior anticancer activity. The cytotoxic effects of CWP and QFF-B proteins are associated with increased production of intracellular ROS and subsequent apoptosis in both cancer cells. MALDI-TOF-MS identified lactophorin, glycation-dependent cell adhesion molecule1 (GlyCAM-1), and its three driven fragments as dominant peptides in QFF-B, while RP-HPLC-P3 contained two of them with molecular masses of 8080.3 and 9395.6 Da. The two peptides, both derived from the C-terminal of lactophorin, were the most representative peptides in the most active protein fractions (QFF-B and RP-HPLC-P3).

The results highlight for the first time that lactophorin is the major anti-cancer ingredient in camel milk and its unique C-terminal peptides present potential candidacy as anticancer agents in nutraceutical and pharmacological applications.

Genistein is an isoflavone that has been reported to have various anti-cancer properties.

This study aimed to reveal whether or not the anti-cancer properties of genistein in AGS gastric cancer cell line were mediated through caspase-3 enzyme.

AGS gastric cancer cells were treated with different concentrations of genistein for 12, 24, and 48 hours and, then, the viability of the cells and IC50 were determined. To determine the effect of genistein on AGS cell migration potency, the wound healing assay was performed. The genistein-induced apoptosis in AGS gastric cells was evaluated by flow cytometry. Caspase-3 gene (CASP3) expression level and its enzyme activity level were determined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and colorimetric techniques, respectively.

The IC50 value was calculated as 70 µM concentration for 24 hours of incubation with genistein. Genistein significantly reduced AGS cell migration compared to the untreated control cells (P<0.001). Genistein increased the early and late apoptosis of the cells (P<0.001) and upregulated the caspase-3 gene expression (P<0.001), but did not significantly enhance the caspase-3 enzyme activity in treated cells.

Genistein exhibited anti-cancer effects on AGS cells to some extent by reducing cellular migration, increasing apoptosis, and upregulating CASP3 gene expression; however, it did not alter the caspse-3 activity. Therefore, it was recommended that more studies should be carried out to delineate the role of caspase-3 in health benefits attributed to the genistein.

Combining various essential oils (EOs) for developing pharmaceutical formulations has been the focus of attention in recent years.

This study aimed to determine the antioxidant effect of the combination of three Eos obtained from clove (Syzygium aromaticum L.), lemon peel (Citrus limon L.), and thyme (Thymus vulgaris L.) by using mixture design.

The EOs of lemon peel (EOL), clove (EOC), and thyme (EOT) as well as their combination were analyzed using a gas chromatograph with flame ionization detector (GC/FID). The antioxidant activities of the EOs from EOL, EOC, and EOT as well as their combination were measured adopting DPPH assay. The construction and statistical analysis of the experiment were designed using the NemrodW (LPRAI, version 2000) software.

EOL, EOC, and EOT were found capable of neutralizing DPPH radical. EOC was distinguished by its strongest antiradical activity with IC50=15.02±0.02 µg/mL. EOT had an IC50=29.20±0.12 µg/mL while EOL had 188.69±0.95 µg/mL. The positive standard BHT was detected to be IC50=24±0.02 µg/ mL. The optimal, combinative mixture of essential oils may have been determined based on these isoresponse curves which allowed fixing the ideal combinations of ingredient in terms of quantity to obtain an EO mixture possessing appreciable and optimal antioxidant characteristics. The predicted antioxidant properties determined by the mixing plan model were retained and the experiments were carried out respecting the contents of proposed ingredients of 25.7% EOT, 32.3% EOL, and 41.9% EOC equivalent to 15.42 mg, 19.38 mg and 25.14 mg, respectively. This resulted in arriving at an essential oil mixture with an experimental IC50=11.023±0.145 µg/mL which was similar to those of the predicted antioxidant properties with an order of 10.907±0.212 µg/mL and a non-significant difference of P<0.05, based on which the validity of the proposed mixing plan model was determined. The combined EO was also found to be rich in eugenol (32.35±1.13%), thymol (25.49±0.03%), and limonene (21.30±0.02%).

Statistical planning and the development of utility profiles for mixtures of essential oils may have been used to predict the optimal composition as well as to determine their antioxidant profile.

This study assessed the effects of combined Funtumia africana and Abutilon mauritianum (CFAAM) leaves on haematological and antioxidant parameters of benign prostate hyperplasia (BPH) induced rats. The study was divided into 5 groups with 6 rats in each group. Group 1 contained the standard control, while group 2 comprised BPH-induced rats that received no treatment. Group 3 had BPH induced rats treated with 5 mg/kg finasteride, whereas groups 4 and 5 consisted of BPH induced rats treated with 200 and 600 mg/kg body weight of CFAAM, respectively. BPH in the rats was induced by daily subcutaneous administration of 5 mg/kg testosterone propionate over a period of 28 days, while treatments with either Finasteride or CFAAM were via the oral route. The obtained results showed a significant (P<0.05) decline in the haemoglobin (Hb), red blood cell (RBC), packed cell volume (PCV), and platelet counts in the BPH-induced untreated rats compared to the normal control. Glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) activities also declined significantly (P<0.05) in the BPH-induced rats compared to the normal control. Further, treatment with CFAAM significantly reversed the observed haematological and antioxidant anomalies in the BPH induced rats, but their total white blood cell (WBC) and differential WBC count values were not significantly altered (P>0.05). These findings suggested that CFAAM maintains a healthy haematological profile and improves the levels of antioxidant parameters under BPH conditions; furthermore, it may be of value in the search for new agents for inhibiting the development and progression of BPH.

Glycation of human serum albumin (HSA) leads to disturbances in its stability, activity, and other properties which, in turn, affect the functional properties of HSA. Modification of albumin by glycation shows considerable potential as a significant biochemical biomarker for diagnosing diabetes. The characteristics of the glycation process in proteins have not been fully examined yet and, therefore, there is insufficient knowledge about them in the field.

This study aimed to clarify the differences between diabetic and non-diabetic HSA as well as their structure-function relationship.

The physiological and laboratory characteristics of glycated albumin as well as HSA were explored. A total of 30 subjects were enrolled in this study in which 15 normal healthy individuals were assigned into the control group, and 15 type-2 diabetic patients were included in the diabetic group. Patients with type-1 diabetes, pregnant women, and individuals with other diseases were excluded from the study. Protein estimation, polyacrylamide gel electrophoresis, ammonium sulphate fractionation, dialysis, glycation of HSA followed by gel electrophoresis of glycated samples, digestion of BSA, as well as HSA by α-chymotrypsin and their documentation and stoichiometry were all performed.

Various characteristic differences were observed between diabetic and non-diabetic HSA including proteolytic susceptibility and in vitro glycation efficiency. Hypoalbuminemia was, particularly, observed in diabetic patients, which was suggestive of a relationship between hyperglycemia and hypoalbuminemia.

Peculiar contrariety between diabetic and non-diabetic HSA, specific differences in their glycation efficiencies, as well as proteolytic susceptibility and their innuendos were precisely traced out. It was concluded that albumin may have been regarded as a significant clinical biomarker for diagnosing diabetes.

An elevated level of uric acid (UA), also known as hyperuricemia (HUA), contributes to the occurrence of cardiovascular diseases (CVDs). However, epidemiolocal features of HUA in populations of Benin are rare.

We identified clinical and metabolic factors associated with HUA in taxi-motorbike drivers (TMDs) of Cotonou.

A total of 134 participants with a mean age of 39.3 years were analyzed using a retrospective cross-sectional study design. Data from a self-administered questionnaire and biochemical markers including, plasma UA, glucose, insulin, creatinine, and lipids were obtained from each participant. HUA was defined as plasma UA greater than 416 μmol/L. Insulin resistance (IR) was determined using the homeostatic model assessment (HOMA). Logistic regression analysis was performed to determine the association of various risk factors with HUA. Odds ratio (OR) and 95% confidence interval (CI) were calculated for HUA.

The overall prevalence of HUA was 19.4% (95% CI: 12.7-26.1) in TMDs. Multivariable logistic regression showed that IR (OR=3.60, 95% CI: 1.27-10.22, P=0.02), hypertension (OR=2.75, 95% CI: 1.00- 7.54, P=0.05), and triglycerides (TG; OR=4.25, 95% CI: 1.39-12.98, P=0.01) were risk factors for HUA. Furthermore, creatinine was inversely associated with HUA (OR=0.62, 95% CI: 0.41-0.94, P=0.02).

HUA was found in 19.4% of the patients. In addition, hypertension, IR, creatinine, and TG levels were independently associated with HUA in TMDs. Therefore, the monitoring of these markers may help prevent HUA.

Dental caries is one of the most common causes threatening human health globally. Sortase A (Srt A) as a transpeptidase, mediates the attachment of the Streptococcus mutans cell wall to dental surfaces by biofilm formation. Due to the development of multidrug-resistance bacteria, attempting to discover growth inhibitors is logical and promising.

The current study aimed at the experimental and docking-based virtual screening of myricetinlike inhibitors for the inhibition of Srt A enzyme in S. mutans isolates.

Sixty-three S. mutans were isolated from pupils based on cultural, morphological, and biochemical characteristics (N=150). After identifying the srtA gene using the polymerase chain reaction (PCR) with specific primers, a broth microdilution test was conducted according to CLSI-2020 criteria to determine the minimum inhibitory concentration (MIC) of myricetin. The in silico exploration of Srt A inhibitors was performed using AutoDock 4.2.6.

The frequency of S. mutans isolates containing the srtA gene was 87.3% of which, fifty isolates (79.4%) were categorized as susceptible to myricetin (MIC,≤16 μg/mL). Of 20 ligands having a high degree of similarity with myricetin, the best docking results were related to ligand 2.

It was concluded that myricetin has an inhibitory effect on oral bacteria in vitro, and ligand 2 had the most negative binding energy (-4.66 kcal/mol) and favorably interacts with the key amino acid residues at the active site of Srt A. Accordingly, this ligand can be utilized as a lead compound for further studies to discover novel inhibitors targeting Srt A in S. mutans.

Nitric oxide (NO) is a signaling molecule that is required for the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). According to previous reports, high concentrations of sodium nitroprusside (SNP) inhibit the osteogenic differentiation of BMSCs, while its low concentration promotes this process.

The present investigation focused on evaluating the underlying mechanism of the osteogenic differentiation of BMSCs treated with low concentrations of SNP as an NO generating agent.

The BMSCs after the 3rd passage was differentiated to osteoblasts when treated with 100 µM for 1 hour every 48 hours until 5, 10, 15, and 20 days of incubation. Then, the matrix production was estimated by quantitative alizarin red assay and calcium determination. The expression of different genes involved in osteogenic differentiation was statistically determined using the reverse transcriptase polymerase chain reaction. Finally, alkaline phosphatase activity was measured by a commercial kit.

The exogenous NO caused a significant (P<0.05) increase in the matrix production of differentiated BMSCs from day 5 to 20. The results showed the elevation of alkaline phosphatase activity and the up-regulation of its gene. Eventually, an increase was observed in the expression of a cascade of other genes such as osteonectin, Bmp7, Smad1, Runx2, and Raf1 in treated BMSCs.

Overall, short-time treatment with a low concentration of exogenous NO increases the matrix production via gene up-regulation and protein production, which might open a new window in treating the low-density bone complication.

Nepeta crispa Willd., a member of Lamiaceae family, is an annual plant native to western Iran, especially Hamedan, with many traditional uses. The plant effects have not been investigated yet. Iron chelating activity is a suitable test for measuring antioxidant activity.

The present study aimed to evaluate Iron-chelating activity of extract, fractions, and essential oil of the N. crispa, in vitro.

The methanolic extract of N. crispa was prepared adopting maceration method. Then, the total methanolic extract was fractionated using hexane, chloroform, ethyl acetate, and water. The essential oil was isolated by hydrodistillation using a Clevenger-type apparatus. Chelating activity of total extract, each fraction and essential oil of N. crispa on Fe2+ ions was determined using Iron-chelating assay. EDTA was employed as positive control.

EDTA as positive control had the best activity with IC50=0.02±0.001 mg/mL, and ascorbic acid came second in this regard (IC50=0.386±0.021 mg/mL). The hexane fraction was the most active fraction among the different fractions of N. crispa (IC50=0.435±0.032 mg/mL). Chelating activity of hexane fraction was followed by aqueous, ethyl acetate, chloroform fractions, and methanolic extract with ICs50 of 1.433±0.098, 2.158±0.074 mg/mL, 3.624±0.112, and 3.051±0.174, respectively. The essential oil showed extremely poor activity in the tested concentrations.

It was concluded that the n-hexane fraction had promising Iron chelating activity, and was likely capable of reducing Fe2+ ions concentration and preventing oxidative damage.

Lactose intolerance is a common pathology that occurs due to the reduced activity of β-galactosidase leaving undigested lactose in the intestine. About 70% of the world population suffers from this condition. The gastro intestinal symptoms associated with this condition are diarrhoea, pain, nausea, bloating, flatulence, etc. It has been reported that these individuals are at a risk of developing several other pathologies like irritable bowel disease, osteoporosis, etc. Hence, proper diagnosis and treatment is essential for dealing with this condition. Various methods are used for providing an accurate diagnosis, such as hydrogen breath test (HBT), lactose intolerance test, genetic test, intestinal biopsy, etc. Depending on the type of intolerance, several methods are adopted for treating it, such as replacing enzyme, using exogenous enzymes, following lactose free diet, as well as consuming prebiotics and probiotics. Different methods are applied to synthesize lactose free dairy products to help lactose intolerant individuals suffering from important vitamins and minerals deprivation. Recently, plant-based milks are also used as a substitute for providing calcium and vitamins. The last few years have seen improvement in the quality and availability of lactose-free dairy products offering tempting foodstuffs to consumers. This narrative article aimed to review the existing science on lactose intolerance, along with its epidemiology, diagnosis, and clinical management.