Unveiling the Chemical Constituents and Inhibitory Roles of Extracts from Pinus pinea L. Nut and Nutshell: A Novel Source for Pharmaceutical Antimicrobials.

Antibiotic resistance in infectious diseases has been a serious problem for the last century, and scientists have focused on discovering new natural antimicrobial agents. Pinus pinea has been used as a natural pharmacotherapeutic agent with antimutagenic, anticarcinogenic, and high antioxidant properties. In this study, GC-MS and LC-HR/MS were employed to analyze Pinus pinea L. nut and nutshell extracts. DPPH radical scavenging assay was performed to analyze the antioxidant properties of the extracts, but no activity was determined. GC-MS analysis showed that linoleic, oleic, and palmitic acids were the three most dominant fatty acids in nut and nutshell extracts, with ratios between 6.75% and 47.06% (v/v). LC-HR/MS revealed that the nutshell methanol extract had a higher phenolic content than other extracts, with vanillic acid (1.4071 mg/g). Antimicrobial activity assays showed that the minimum inhibitory concentrations (MIC) of the extracts varied between 5.94 and 190 mg/mL, and the most significant inhibition was seen in the nutshell methanol extract (MICs: between 5.94 and 47.5 mg/mL). Consequently, the antimicrobial activity of the extracts can be attributed to the dense fatty acids they contain, and the nutshell methanol extract showed the most potent inhibition related to the abundance of phenolic compounds in the extract.

Comparative evaluation of cytotoxic and anti-metastatic function of microbial chondroitin sulfate and animal-originated commercial chondroitin sulfate in cancer cells.

Cancer has the second-highest mortality rate worldwide after cardiovascular disease. In addition, breast and cervical cancer are two of the leading causes of cancer-related deaths among women. The tumor microenvironment, which consists of fibroblasts, immune cells, cells that form blood vessels, and proteins, is a therapeutic target for cancer therapy. As part of the cellular microenvironment, glycosaminoglycan chondroitin sulfate is associated with various aspects of tumor progression and metastasis depending on the sulfate pattern of chondroitin sulfate. This study evaluated the roles of Microbial Chondroitin Sulfate (CS) and Commercial CS in tumor growth and metastasis comparatively using MDA-MB-231 metastatic breast cancer cells, HeLa cervical cancer cells, and normal fibroblasts. In addition, the role of CS types in wound healing was also assessed comparatively.  Microbial CS was more cytotoxic in MDA-MB-231 cells than HeLa compared to Commercial CS. Although both CS reduced cell viability in normal cells, the selective index of Microbial CS in MDA-MB-213 cells was higher than its commercial counterpart. In addition, the role of CS types in wound healing was also assessed comparatively. Both types of CS decreased the cell migration in MDA-MB-231 cancer cells, but HeLa cells were more sensitive to Microbial CS than Commercial CS to heal the wound. The wound healing of NIH3T3 cells after Microbial CS was similarly high to the healing after Commercial CS. This preliminary study shows that microbial CS produced by biotechnological methods from a recombinant source created by our team can be an effective therapeutic agent in various types of cancer.

Comparative Analysis of Antioxidant, Anticholinesterase, and Antibacterial Activity of Microbial Chondroitin Sulfate and Commercial Chondroitin Sulfate.

Chondroitin synthesis was performed using the recombinant Escherichia coli(C2987) strain created by transforming the plasmid pETM6-PACF-vgb, which carries the genes responsible for chondroitin synthesis, kfoA, kfoC, kfoF, and the Vitreoscilla hemoglobin gene (vgb). Then, Microbial chondroitin sulfate (MCS)'s antioxidant, anticholinesterase, and antibacterial activity were compared with commercial chondroitin sulfate (CCS). The antioxidant studies revealed that the MCS and CCS samples could be potential targets for scavenging radicals and cupric ion reduction. MCS demonstrated better antioxidant properties in the ABTS assay with the IC50 value of 0.66 mg than CCS. MCS showed 2.5-fold for DPPH and almost 5-fold for ABTS⋅+ (with a value of 3.85 mg/mL) better activity than the CCS. However, the compounds were not active for cholinesterase enzyme inhibitions. In the antibacterial assay, the Minimum inhibitory concentration (MIC) values of MCS against S. aureus, E. aerogenes, E. coli, P. aeruginosa, and K. pneumoniae (0.12, 0.18, 0.12, 0.18, and 0.18 g/mL, respectively) were found to be greater than that of CCS (0.42, 0.48, 0.36, 0.36, and 0.36 g/mL, respectively). This study demonstrates that MCS is a potent pharmacological agent due to its physicochemical properties, and its usability as a therapeutic-preventive agent will shed light on future studies.

The test-retest reliability and minimal clinically important difference of the Dubousset Functional Test and its correlation with Rolland Morris disability questionnaire in chronic non-specific low back pain.

OBJECTIVE: This study examines the test-retest reliability, the minimal clinically important difference (MCID), and its correlation with the Rolland Morris Disability Questionnaire (RMDQ) of the Dubousset Functional Test (DFT) in evaluating the functional capacity and dynamic balance of patients with chronic non-specific low back pain (cnsLBP). METHODS: Seventy-five patients with cnsLBP aged 18 years and over were included. The Five-Repetition Sit-To-Stand Test (5R-STS), the subcomponents of the DFT (the Up and Walk Test, the Steps Test, the Down and Sitting Test, and the Dual-Tasking Test) were administered to the patients. Patients were rested for 1 h, and the DFT was applied again. Pain level was evaluated with the Visual Analogue Scale before the tests started and after the tests were completed. Self-report function assessment was made using the RMDQ. RESULTS: The test-retest reliability of the subcomponents of the DFT was excellent. The ICCs were: 0.91, 0.86, 0.89, and 0.89, respectively. The standard measurement errors of the subcomponents of the DFT were 0.32, 0.12, 0.14, and 0.25, respectively. The subcomponents of the DFT were highly correlated with the RMDQ and 5R-STS with the correlation coefficients of 0,83, 0,83, 0,79, 0,83 and 0,81, 0,75, 0,73, and 0,82, respectively (p < 0.01). The MCIDs of the subcomponents were 0,60, 0,23, 0,27, and 0,48, respectively. CONCLUSION: The DFT is reliable in evaluating patients' functional capacity and dynamic balance with cnsLBP without causing discomfort. It is simple, quick, and simultaneously assesses multiple areas contributing to spinal alignment, muscle integrity, and balance.

Rapid identification of Gram-positive pathogens and their resistance genes from positive blood culture broth using a multiplex tandem RT-PCR assay.

The early initiation of targeted antibiotic therapy in patients with bacteraemia and septic shock impacts favourably on outcomes. Rapid methods are therefore increasingly employed for bacterial identification directly from positive blood culture bottles, but with variable success. We evaluated the performance of the Gram Positive 12 multiplex tandem PCR (MT-PCR) assay (AusDiagnostics; catalogue no. 6202, version 07) containing targets for the identification of staphylococci including Staphylococcus aureus, streptococci including Streptococcus pneumoniae, enterococci including Enterococcus faecalis and Enterococcus faecium and their common antibiotic resistance genes (mecA, vanA, vanB). A total of 673 aerobic and anaerobic blood culture broths demonstrating Gram-positive cocci on microscopy were analysed in parallel with traditional phenotypic methods. Amplification of the internal control was inhibited in 79/673 (11.7 %) samples; however, MT-PCR identification was in concordance with phenotypic identification to the genus level in 96.6 % (537/556) of the remaining monomicrobial specimens and to the species level, where applicable, in 100 % (172/172) of samples. MT-PCR identification for 94.7 % (36/38) of polymicrobial samples matched traditional phenotypic identification. Meticillin and vancomycin susceptibility results determined by MT-PCR in blood culture broths demonstrated complete agreement with those determined by phenotypic methods in all 143 Staphylococcus aureus isolates and eight E. faecium isolates, respectively. Gram-positive pathogens and their key antibiotic resistance markers were reliably identified with the MT-PCR assay within 3 h of a positive blood culture result.