Characterization of <i>Lacticaseibacillus rhamnosus</i>, <i>Levilactobacillus brevis</i> and <i>Lactiplantibacillus plantarum</i> Metabolites and Evaluation of Their Antimicrobial Activity against Food Pathogens

Lactic acid bacteria (LAB) play an important role as natural food preservatives. However, the characterization of the variety of their metabolites is limited. The objective of this study was to determine the production of specific metabolites of Lacticaseibacillus rhamnosus, Levilactobacillus brevis and Lactiplantibacillus plantarum by an optimized liquid chromatography with an ultraviolet/diode detection (HPLC-UV/DAD) method and to investigate their potential antimicrobial activity against specific food pathogens. Based on the results of this study, the main metabolites detected in Levilactobacillus brevis were 103.4 μg mL−1 DL-p-Hydroxyphenyllactic acid (OH-PLA) and 2.59 μg mL−1 vanillic acid, while 216.2 μg mL−1 OH-PLA, 19.0 μg mL−1 salicylic acid, 3.7 μg mL−1 vanillic acid, 6.9 μg mL−1 ferulic acid, 4.2 μg mL−1 benzoic acid and 1.4 μg mL−1 4-Hydrocinnamic acid were identified in the Lactiplantibacillus plantarum strain and 147.6 μg mL−1 OH-PLA and 4.9 μg mL−1 ferulic acid were identified in Lacticaseibacillus rhamnosus. This study provides alternative approaches for the molecules involved in the antimicrobial activity of food microorganism fermentation. These molecules may be used as antimicrobial ingredients in the food industry instead of conventional chemical preservatives

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Mutational Pattern in the Fourth Pandemic Phase in Greece

The aim of this study is to investigate the circulating variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from Athens and from rural areas in Greece during July and August 2021. We also present a rapid review of literature regarding significant SARS-CoV-2 mutations and their impact on public health. A total of 2500 nasopharyngeal swab specimens were collected from suspected COVID-19 cases (definition by WHO 2021b). Viral nucleic acid extraction was implemented using an automatic extractor and the RNA recovered underwent qRT-PCR in order to characterize the specimens as positive or negative for SARS-CoV-2. The positive specimens were then used to identify specific Spike gene mutations and characterize the emerging SARS-CoV-2 variants. For this step, various kits were utilized. From the 2500 clinical specimens, 220 were tested positive for SARS-CoV-2 indicating a prevalence of 8.8% among suspected cases. The RT-PCR Ct (Cycle threshold) Value ranged from 19 to 25 which corresponds to medium to high copy numbers of the virus in the positive samples. From the 220 positive specimens 148 (67.3%) were from Athens and 72 (32.7%) from Greek rural areas. As far as the Spike mutations investigated: N501Y appeared in all the samples, D614G mutation appeared in 212 (96.4%) samples with a prevalence of 87.2% in Athens and 98.6% in the countryside, E484K had a prevalence of 10.8% and 12.5% in Athens and the rural areas, respectively. K417N was found in 18 (12.2%) samples from Athens and four (5.6%) from the countryside, P681H was present in 51 (34.5%) Athenian specimens and 14 (19.4%) specimens from rural areas, HV69-70 was carried in 32.4% and 19.4% of the samples from Athens and the countryside, respectively. P681R had a prevalence of 87.2% in Athens and 98.6% in rural areas, and none of the specimens carried the L452R mutation. 62 (28.2%) samples carried the N501Y, P681H, D614G and HV69-70 mutations simultaneously and the corresponding variant was characterized as the Alpha (UK) variant (B 1.1.7). Only six (2.7%) samples from the center of Athens had the N501Y, E484K, K417N and D614G mutations simultaneously and the virus responsible was characterized as the Beta (South African) variant (B 1.351). Our study explored the SARS-CoV-2 variants using RT-PCR in a representative cohort of samples collected from Greece in July and August 2021. The prevalent mutations identified were N501Y (100%), D614G (96.4%), P681R (90.1%) and the variants identified were the Delta (90.1%), Alpha (28.2%) and Beta (2.7%)

Application of microfluidic paper-based analytical devices (μPADs) for food microbial detection.

Microfluidic technology is an emerging scientific area, aiming to provide inexpensive, sensitive and efficient devices which can, among other applications, may play a significant role in food monitoring. Microfluidic Paper-based analytical Devices (μPADs) have been proposed as potentially effective tools for safety and quality monitoring, in terms of microbial detection. Several μPADs have been developed using alternative fabrication methods, each one exhibiting different features, advantages and limitations. These devices have been tested for their applicability for the detection of foodborne pathogens in cultured cells, water or food matrices. Limited studies report the utilization of μPADs for quality monitoring of food, targeting microbial or chemical quality indexes. The aim of the article is to describe and discuss the most common types of μPADs for their effectiveness in microbial detection in food matrices and thus theirs applicability in the food industry sector as simple and cost effective tools for rapid safety and quality monitoring. This article is protected by copyright. All rights reserved

Comparison of Q223R leptin receptor polymorphism to the leptin gene expression in Greek young volunteers Running title: Leptin polymorphism and leptin gene expression

Objective: The objective of the present study was to identify the leptin gene expression and the leptin receptor polymorphisms in blood samples and to correlate gene expression values with an-thropometric characteristics. Methods: Blood from 140 Greek young volunteers was subjected to pol-ymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP), for the genomic re-gion of Q223R polymorphism at codon 223 in the leptin receptor gene (LEPR) coding region. RNA extraction, cDNA synthesis and Quantitative Real-Time PCR was performed for assessing the expres-sion of the leptin gene (LEP). Results: Leptin gene was identified in all tested specimens and the gene was expressed in 88.9% of all volunteers with BMI &lt; 25. In addition, it was observed that gene ex -pression is affected by various external factors, such as Body Mass Index (BMI), eating behavior, gender and age. It was also shown that as for the Q223R polymorphism (A to G) allele G occurs with a frequency of 100% in men with BMI &gt; 30 and 75.9% in men and 88.9% in women with BMI 25-30.Volunteers with BMI 25-30 who were homozygous on the G allele were 50% and 77.8% in men and women respectively. All subjects with a BMI &gt; 30 were homozygous on the G allele at 100%. Conclusions: In this small-scale study, results have shown that the leptin gene expression correlates with BMI and that the allele G in Q223R polymorphism is linked to overweight individuals

The Impact of Obesity on the Fitness Performance of School-Aged Children Living in Rural Areas&mdash;The West Attica Project

This study aimed to investigate the relationship of body mass index (BMI) with muscle and cardiorespiratory fitness in children living within rural areas (regional unit of West Attica) in Greece. Participants included 399 students (187 boys, 212 girls), ages 8&ndash;12 years old, and were evaluated in physical performance tests. The point prevalence of overweight and obesity was 21.39% and 26.20% in boys, and 19.90% and 23.79% in girls. Significant differences were observed in all physical performance tests (handgrip, long jump, shuttle run, trunk flexors, and extensors endurance) between normal weight and obese participants. BMI was positively correlated with handgrip (r = 0.442&minus;0.462, p &lt; 0.001). There was a negative association with long jump (r = &minus;0.206, p &lt; 0.001), 20 m shuttle run (r = &minus;0.394, p &lt; 0.001), trunk flexors (r = &minus;0.403, p &lt; 0.001) and trunk extensors endurance (r = &minus;0.280, p &lt; 0.001). The regression analysis showed that 20&ndash;30% of the overall variation for physical performance assessments could be accounted for by BMI, age, and sex. With the exception of the long jump and the endurance of the trunk extensors, BMI alone may explain more than 10% of the outcome of most tests. This study highlights the determinant of BMI on muscle and cardiorespiratory fitness. The management of obesity should begin early in childhood to prevent adult chronic cardiovascular and metabolic diseases

Association of <i>TNF</i>-α 308G/A and <i>LEPR</i> Gln223Arg Polymorphisms with the Risk of Type 2 Diabetes Mellitus

The objective of the present study was to identify the association of the TNF-α- 308G/A and leptin receptor (LEPR) Gln223Arg polymorphisms with the risk of development of type 2 diabetes mellitus (T2DM). Methods: A total of 160 volunteers were studied: 108 with T2DM and 52 participants as control, who served as the control group. Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) for the genomic region of TNF-α- 308G/A and LEPR Gln223Arg were carried out. Results: The frequency of LEPR Gln223Arg genotypes in T2DM and control groups showed significant differences in the distribution of genotypes (p TNF-α- 308G/A genotypes in T2DM and control subjects showed significant differences in the distribution of genotypes (p p < 0.05)

Platelet, Fibrinolytic and Other Coagulation Abnormalities in Newly-Diagnosed Patients with Chronic Thromboembolic Pulmonary Hypertension

The pathophysiological background of chronic thromboembolic pulmonary hypertension (CTEPH) has not been fully elucidated. Evidence suggests that abnormal platelet function and ineffective fibrinolysis may play a key role in the development of the disease. The purpose of this study was to evaluate platelet and coagulation function in CTEPH, using non-conventional global coagulation assays, and platelet activation and endothelial dysfunction laboratory markers. A total of 40 newly-diagnosed CTEPH patients were studied, along with 35 healthy controls. Blood samples from CTEPH patients were taken directly from the pulmonary artery. All subjects were assessed with platelet function analyzer-100, light transmission aggregometry, thromboelastometry, endogenous thrombin potential. von Willebrand antigen and activity, p-selectin, thromboxane A2 and serotonin levels were also assessed. The results showed that CTEPH patients present diminished platelet aggregation, presence of disaggregation, decreased rate of fibrinolysis, defective thrombin generation and increased levels of thromboxane A2, p-selectin, von Willebrand antigen and activity. Serotonin levels did not present any differences between the two groups. The results of this study suggest that CTEPH patients present platelet function, fibrinolytic, thrombin generation and other clot formation abnormalities. Well-designed clinical studies are needed to further evaluate the complex hemostatic abnormalities in the CTEPH setting and assess their potential clinical applications

&lt;p&gt;Association of TNF-a 308G/A and LEPR Gln223Arg Polymorphisms with the Risk of Type 2 Diabetes Mellitus&lt;/p&gt;

The objective of the present study was to identify the association of the TNF-alpha- 308G/A and leptin receptor (LEPR) Gln223Arg polymorphisms with the risk of development of type 2 diabetes mellitus (T2DM). Methods: A total of 160 volunteers were studied: 108 with T2DM and 52 participants as control, who served as the control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for the genomic region of TNF-alpha- 308G/A and LEPR Gln223Arg were carried out. Results: The frequency of LEPR Gln223Arg genotypes in T2DM and control groups showed significant differences in the distribution of genotypes (p &lt; 0.05). The frequency also of TNF-alpha- 308G/A genotypes in T2DM and control subjects showed significant differences in the distribution of genotypes (p &lt; 0.05). Conclusion: Our results indicate that there are significant differences in the distribution of genotypes and alleles between the individuals with T2DM and control subjects (p &lt; 0.05)

Identification of Polymorphisms of the <i>CSN2</i> Gene Encoding β-Casein in Greek Local Breeds of Cattle

This e research focused on the detection and identification of genetic polymorphisms in exon 7 of the β-casein CSN2 gene in blood samples from Greek Holstein cows and from local breeds of cattle, such as Vrachykeratiki, Katerinis, and Sykias. For this purpose, DNA was isolated from 780 blood samples obtained from Greek Holstein cows, 86 from three local breeds of cattle, namely Brachyceros, Katerinis, and Sykias, and 14 from Greek buffalo. The desired region of exon 7 was amplified by PCR, resulting in 121 and 251 bp products in bovine and buffalo samples. The PCR product was digested with restriction fragment length polymorphism (RFLP) on agarose gels. The restriction enzymes DdeI and TaqI were used. All of the blood samples had the amplified size. The results showed that 74.4% of the Greek Holstein cows had the A2A2 β-casein genotype, the three native breads Vrachykeratiki had 57.7%, and the other two had 100% of the A2A2 β-casein. From the 14 Greek buffalo, 100% had the A2A2 β-casein