Comparative Probiogenomics Analysis of Limosilactobacillus fermentum 3872.

Limosilactobacillus fermentum is an important member of the lactic acid bacteria group and holds immense potential for probiotic properties in human health and relevant industries. In this study, a comparative probiogenomic approach was applied to analyze the genome sequence of L. fermentum 3872, which was extracted from a commercially available yogurt sample, along with 20 different publicly available strains. Results indicate that the genome size of the characterized L. fermentum 3892 strain is 2,057,839 bp, with a single- and circular-type chromosome possessing a G + C content of 51.69%. The genome of L. fermentum 3892 strain comprises a total of 2120 open reading frames (ORFs), two genes encoding rRNAs, and 53 genes encoding tRNAs. Upon comparative probiogenomic analysis, two plasmid sequences were detected among the study strains, including one for the L. fermentum 3872 genome, which was found between position 1,288,203 and 1,289,237 with an identity of 80.98. The whole-genome alignment revealed 2223 identical sites and a pairwise identity of 98.9%, indicating a significant difference of 1.1% among genome strains. Comparison of amino acid encoding genes among strains included in this study suggests that the strain 3872 exhibited the highest degree of amino acids present, including glutamine, glutamate, aspartate, asparagine, lysine, threonine, methionine, and cysteine. The comparative antibiotic resistome profiling revealed that strain 3872 exhibited a high resistant capacity only to ciprofloxacin antibiotics as compared to other strains. This study provides a genomic-based evaluation approach for comparative probiotic strain analysis in commercial foods and their significance to human health.

Promising phytopharmacology, nutritional potential, health benefits, and traditional usage of Tribulus terrestris L. herb.

Traditional medicines are becoming more popular as people become more aware of the dangers of synthetic pharmaceuticals. Tribulus terrestris L., (Gokharu) an annual herbaceous plant, has been extensively utilized by herbalists for numerous medicinal purposes. T. terrestris has been studied for its multiple therapeutic effects, including immunomodulatory, aphrodisiac, anti-urolithic, absorption enhancer, cardioprotective, antidiabetic, anti-inflammatory, hypolipidemic, neuro-protective, anticancer, and analgesic properties. Saponins and flavonoids are two examples of beneficial substances that have recently been found in T. terrestris. These chemicals are very important for a variety of therapeutic effects. Numerous studies have shown that T. terrestris products and various parts may have antioxidant, anti-inflammatory, anti-cancer, anti-diabetic, testosterone-boosting, and liver protective effects. According to the published evidence, T. terrestris boosts testosterone secretion, regulates blood pressure, and protects the human body against injuries. The cardiovascular, reproductive, and urinary systems are all severely impacted. Due to its potent bioactive compounds, the literature evaluated from a wide range of sources including books, reports, PubMed, ScienceDirect, Wiley, Springer, and other databases demonstrated the extraordinary potential to treat numerous human and animal ailments. Our review is different from other published articles because we explored its importance for humans and especially in veterinary like poultry health. It could also be used as an aphrodisiac to treat different fertility-related disorders in human and animal science. More research into the pharmacodynamics of herbs like T. terrestris is needed so that it can be used in a wider variety of nutraceutical products for humans and poultry.

The Impact of Flap Vascular Pressurization Technique on the Repair of Large-Area Soft Tissue Defects in Limbs.

Objective: This study investigated the impact of the flap vascular pressurization technique on repairing large-area soft tissue defects in the limbs. Methods: This study employed a randomized controlled trial design to enroll patients with large-area skin defects in the limbs, accompanied by exposed deep tissues such as nerves, blood vessels, bones, and tendons, for various reasons between July 2020 to July 2022. The patients were randomly assigned into two groups using a random number table method. The control group (n = 30) underwent traditional anterior lateral thigh flap repair, while the experimental group (n = 30) underwent flap repair using the vascular pressurization technique. Clinical indicators, flap survival, scar formation, and satisfaction were compared between the two groups. Results: There were no significant differences in operation time, intraoperative blood loss, and length of hospital stay between the two groups (P > .05). The flap survival rate in the experimental group (90.00%, 27/30) was significantly higher than that in the control group (66.67%, 20/30) (P < .05). The Manchester Scar Scale (MSS) scores in the experimental group were significantly higher than those in the control group (P < .05). The satisfaction rate in the experimental group (93.33%, 28/30) was significantly higher than that in the control group (73.33%, 22/30) (P < .05). Conclusion: The use of the flap vascular pressurization technique for the repair of soft tissue defects in the limbs can significantly increase flap survival rate, improve scar formation, and enhance patient satisfaction, thereby demonstrating good clinical value. The flap vascular pressurization technique can be promoted as a reliable method for repairing large-area skin defects in the limbs, thereby contributing to the advancement of specialized fields.

Snapshot of the Probiotic Potential of Kluveromyces marxianus DMKU-1042 Using a Comparative Probiogenomics Approach.

Kluyveromyces marxianus is a rapidly growing thermotolerant yeast that secretes a variety of lytic enzymes, utilizes different sugars, and produces ethanol. The probiotic potential of this yeast has not been well explored. To evaluate its probiotic potential, the yeast strain Kluyveromyces marxianus DMKU3-1042 was analyzed using next-generation sequencing technology. Analysis of the genomes showed that the yeast isolates had a GC content of 40.10-40.59%. The isolates had many genes related to glycerol and mannose metabolism, as well as genes for acetoin and butanediol metabolism, acetolactate synthase subunits, and lactic acid fermentation. The strain isolates were also found to possess genes for the synthesis of different vitamins and Coenzyme A. Genes related to heat and hyperosmotic shock tolerance, as well as protection against reactive oxygen species were also found. Additionally, the isolates contained genes for the synthesis of lysine, threonine, methionine, and cysteine, as well as genes with anticoagulation and anti-inflammatory properties. Based on our analysis, we concluded that the strain DMKU3-1042 possesses probiotic properties that make it suitable for use in food and feed supplementation.

Cervical Range of Motion after Anterior Corpectomy and Fusion versus Laminoplasty for Multilevel Cervical Spondylotic Myelopathy: A Three-Dimensional Comparison Based on the Coda Motion System.

OBJECTIVE: To compare the change in cervical range of motion (CROM) after the 2 most widely used techniques for multilevel cervical spondylotic myelopathy (CSM): anterior cervical corpectomy and fusion (ACCF) and laminoplasty. METHODS: Patients with multilevel CSM treated in our hospital between 2014 and 2018 were divided into an ACCF group and a laminoplasty (LAMP) group according to the treatment received. Their demographic data, preoperative and postoperative Japanese Orthopedic Association (JOA) scores, and CROM, measured using the Coda Motion system, were analyzed and compared. RESULTS: A total of 53 patients were enrolled, including 29 patients in the ACCF group and 24 patients in the LAMP group. Age, sex, duration of follow-up, and preoperative and postoperative JOA scores were comparable in the 2 groups. Compared with preoperative measurements, ACCF group lost an average of 9.8%, 28.5%, 8.9%, 9.9%, 10.6%, and 7.8% of their CROM in flexion, extension, left and right lateral flexion, and left and right rotation, respectively, at the latest follow-up. For the LAMP group, these average losses were 3.5%, 16.4%, 3.2%, 6.3%, 7.0%, and 5.7%, respectively. Thus, the ACCF group exhibited greater average CROM loss than the LAMP group in all directions at the latest follow-up. CONCLUSIONS: Both ACCF and laminoplasty cause significant CROM loss in patients with multilevel CSM. The laminoplasty technique preserved more CROM than ACCF in all 6 directions after at least 1 year of follow-up. These results can be used when counseling patients undergoing surgery.

Hyper-osmolarity environment-induced oxidative stress injury promotes nucleus pulposus cell senescence in vitro.

Nucleus pulposus (NP) cell senescence is involved in disc degeneration. The in situ osmolarity within the NP region is an important regulator of disc cell's biology. However, its effects on NP cell senescence remain unclear. The present study was aimed to investigate the effects and mechanism of hyper-osmolarity on NP cell senescence. Rat NP cells were cultured in the in situ-osmolarity medium and hyper-osmolarity medium. The reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) was added along with the medium to investigate the role of oxidative injury. Cell cycle, cell proliferation, senescence associated ß-galactosidase (SA-ß-Gal) activity, telomerase activity, expression of senescence markers (p16 and p53) and matrix molecules (aggrecan and collagen II) were tested to assess NP cell senescence. Compared with the in situ-osmolarity culture, hyper-osmolarity culture significantly decreased cell proliferation and telomerase activity, increased SA-ß-Gal activity and cell fraction in the G0/G1 phase, up-regulated expression of senescence markers (p16 and p53) and down-regulated expression of matrix molecules (aggrecan and collagen II), and increased intracellular ROS accumulation. However, addition of NAC partly reversed these effects of hyper-osmolarity culture on cellular senescence and decreased ROS content in NP cells. In conclusion, a hyper-osmolarity culture promotes NP cell senescence through inducing oxidative stress injury. The present study provides new knowledge on NP cell senescence and helps us to better understand the mechanism of disc degeneration.

Impact of orientation and flexibility of peptide linkers on T. maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner.

Fusion protein construction often requires peptide linkers for prolonged conformation, extended stability and enzyme activity. In this study a series of fusion between Thermotoga maritima lipase Tm1350 and Bacillus subtillis coat protein CotB, comprising of several peptide linkers, with different length, flexibility and orientations were constructed. Effects of temperature, pH and chemicals were examined, on the activity of displayed enzyme. The fusion protein with longer flexible linkers L9 [(GGGGS)4] and L7 (GGGGS-GGGGS-EAAAK-EAAAK-GGGGS-GGGGS) possess 1.29 and 1.16-fold higher activity than the original, under optimum temperature and pH respectively. Moreover, spore surface displaying Tm1350 with L3 (EAAAK-GGGGS) and L9 ((GGGGS)4) showed extended thermostably, maintaining 1.40 and 1.35-fold higher activity than the original respectively, at 80 °C after 5 h of incubation. The enzyme activity of linkers with different orientation, including L5, L6 and L7 was determined, where L7 maintained 1.05 and 1.27-fold higher activity than L5 and L6. Effect of 0.1% proteinase K, bromelain, 20% ethanol and 30% methanol was investigated. Linkers with appropriate Glycine residues (flexible) showed higher activity than Alanine residues (rigid). The activity of the displayed enzyme can be improved by maintaining orientation and flexibility of peptide linkers, to evaluate high activity and stability in industrial processes.

Expression and Characterization of a Novel Thermo-Alkalistable Lipase from Hyperthermophilic Bacterium Thermotoga maritima.

A gene coding for lipase (Tm1350) from the hyperthermophilic bacterium Thermotoga maritima MSB8 was cloned and overexpressed by Escherichia coli. The enzyme can degrade substrates with both short and long acyl chain lengths. The apparent Km and Vmax values for p-nitrophenyl butyrate were 8 mM and 333 U/mg, respectively. The enzyme displayed optimal activity at pH 7.5 and 70 °C, maintained 66 % of the original activity after 8 h of incubation, and its half-lives at pHs 9 and 10 were 8 and 1 h. The activity of Tm1350 was stimulated up to 131 or 151 % of the original activity by incubating with 4 M urea or 20 % (v/v) methanol, and 90.1 or 70.2 % of the activity was maintained after 8 h incubation of the enzyme in 20 or 75 % (v/v) of the methanol, showing potential for biodiesel production. The activity of the enzyme without cysteine residue was stimulated up to 618 and 550 % of the original activity by incubating with dithiothreitol (DTT) and reduced glutathione (GSH) at a concentration of 1 mM. However, the circular dichroism spectra of the enzyme have no obvious change after DTT treatment. It is speculated that DTT interacts with potential residues in some key active sites without influence of structure.

Characterization of ATPase activity of class II chaperonin from the hyperthermophilic archaeon Pyrococcus furiosus.

To understand how molecular damage under harsh environmental conditions can be controlled, we investigated the properties of ATPase activity of the chaperonin molecular machinery from the hyperthermophilic archaeon Pyrococcus furiosus (PfCPN). PfCPN ATPase activity depended on K(+) and Mg(2+) and its optimal pH was 7.5. PfCPN had almost no ADPase activity. ADP strongly competitively inhibited PfCPN ATPase activity. Inhibition of PfCPN ATPase decreased its chaperonin activity in protecting lysozyme from heat-induced inactivation.

Expression and characterization of the chaperonin molecular machine from the hyperthermophilic archaeon Pyrococcus furiosus.

The chaperonin molecular machine from hyperthermophilic archaeon Pyrococcus furiosus was studied in this paper. The Pyrococcus furiosus chaperonin gene (PfCPN) was amplified by PCR from the Pyrococcus furiosus genomic DNA, and expressed in Escherichia coli BL21-Codonplus(DE)(3)-RIL. The recombinant PfCPN was purified to homogeneity by using ion-exchange and size-exclusion chromatography. It was found that the ATPase activity of the PfCPN was highest at 88 degrees C, and there existed a nested cooperativity of the ATPase activity of the PfCPN. This result suggested that nested allosteric behavior may be common to chaperonin molecular machines from archaea. The half-life (t(1/2)) of the ATPase activity of the PfCPN at 100 degrees C was about 60 min. The PfCPN displayed chaperone activity in preventing lysozyme from thermal inactivation. This chaperone activity was in an ATP-dependent manner.

Angiopoietin-related growth factor (AGF) supports adhesion, spreading, and migration of keratinocytes, fibroblasts, and endothelial cells through interaction with RGD-binding integrins.

Angiopoietin-related growth factor (AGF) is a newly identified member of angiopoietin-related proteins (ARPs)/angiopoietin-like proteins (Angptls). AGF has been considered as a novel growth factor in accelerating cutaneous wound healing, as it is capable of stimulating keratinocytes proliferation as well as angiogenesis. But in our paper, we demonstrate that AGF stimulates keratinocytes proliferation only at high protein concentration, however, it can potently promote adhesion, spreading, and migration of keratinocytes, fibroblasts, and endothelial cells. Furthermore, we confirm that the adhesion and migration cellular events are mediated by RGD-binding integrins, most possibly the alpha(v)-containing integrins, by in vitro inhibition assays using synthetic competitive peptides. Our results strongly suggest that AGF is an integrin ligand as well as a mitogenic growth factor and theoretically participates in cutaneous wound healing in a more complex mechanism.