Red biocolorant from endophytic Talaromyces minnesotensis: production, properties, and potential applications.

Fungal colorants are gradually entering the global color market, given their advantages of being less harmful to human health, as well as having greater stability and biotechnological potential, compared to other natural sources. The present work concerns the isolation and identification of an endophytic filamentous fungus, together with the chemical characterization and assessment of the fluorescence, toxicity, stability, and application potential of its synthesized red colorant. The endophytic fungus was isolated from Hymenaea courbaril, a tree from the Brazilian savannah, and was identified as Talaromyces minnesotensis by phenotypic and genotypic characterization. Submerged cultivation of the fungus resulted in the production of approximately 12 AU500 of a red biocolorant which according to LC-DAD-MS analysis is characterized by being a complex mixture of molecules of the azaphilone class. Regarding cytotoxicity assays, activity against human hepatoblastoma (HepG2) cells was only observed at concentrations above 5.0 g L-1, while antimicrobial effects against pathogenic bacteria and yeast occurred at concentrations above 50.0 g L-1. The biocolorant showed high stability at neutral pH values and low temperatures (10 to 20 °C) and high half-life values (t1/2), which indicates potential versatility for application in different matrices, as observed in tests using detergent, gelatin, enamel, paint, and fabrics. The results demonstrated that the biocolorant synthesized by Talaromyces minnesotensis has potential for future biotechnological applications. KEY POINTS: • An endophytic fungus, which was isolated and identified, synthesize a red colorant. • The colorant showed fluorescence property, low toxicity, and application potential. • The red biocolorant was highly stable at pH 8.0 and temperatures below 20°C.

Purification and characterization of two new antimicrobial molecules produced by an endophytic strain of Paenibacillus polymyxa.

An endophytic bacterium inhibiting pathogenic bacteria was isolated and the strain was genetically identified as Paenibacillus polymyxa. Biochemical characterization of fermentation broth indicated the presence of peptidic antimicrobial molecules. Liquid-liquid partition resulted in an organic fraction (OF) and an aqueous fraction (AF). OF presented a broad spectrum of activity against a panel of pathogenic bacteria and a fungus whereas the AF was active only against Gram-negative bacteria. AF was sequentially submitted to ion-exchange, desalting and reverse phase (RP) chromatography. A molecule with an RT of 2.45 min exhibited activity against all Gram-negative pathogenic strains tested beside P. mirabilis. The primary structure of the molecule, named AMP-Pp, was determined as Gly-Glu-Hyp-Gly-Ala by N-terminal sequencing. The molecular mass and amino acid sequence were confirmed by MS/MS. With a molecular mass of 463 Da, AMP-Pp is one of the smallest active natural peptides reported, yet. RP chromatography of OF resulted in four peaks. The first three peaks corresponded to known antimicrobials. MS analysis of peak 4 revealed the presence of an ion with m/z 3,376.4 Da, whose proposed molecular formula is C182H321N29O29. The compound, named polycerradin, showed a spectrum of activity against Gram-positive bacteria, Gram-negative bacteria (beside P. mirabilis) and a fungus.

Bone turnover markers in sheep and goat: A review of the scientific literature.

Bone turnover markers (BTMs) are product of bone cell activity and are generally divided in bone formation and bone resorption markers. The purpose of this review was to structure the available information on the use of BTMs in studies on small ruminants, especially for monitoring their variations related to diet, exercise, gestation and metabolic lactation state, circadian and seasonal variations, and also during skeletal growth. Pre-clinical and translational studies using BTMs with sheep and goats as animal models in orthopaedic research studies to help in the evaluation of the fracture healing process and osteoporosis research are also described in this review. The available information from the reviewed studies was systematically organized in order to highlight the most promising BTMs in small ruminant research, as well as provide a wide view of the use of sheep and goat as animal models in orthopaedic research, type of markers and commercial assay kits with cross-reactivity in sheep and goat, method of sample and storage of serum and urine for bone turnover markers determination and the usefulness and limitations of bone turnover markers in the different studies, therefore an effective tool for researchers that seek answers to different questions while using BTMs in small ruminants.

Bone turnover markers for early detection of fracture healing disturbances: A review of the scientific literature.

Imaging techniques are the standard method for assessment of fracture healing processes. However, these methods are perhaps not entirely reliable for early detection of complications, the most frequent of these being delayed union and non-union. A prompt diagnosis of such disorders could prevent prolonged patient distress and disability. Efforts should be directed towards the development of new technologies for improving accuracy in diagnosing complications following bone fractures. The variation in the levels of bone turnover markers (BTMs) have been assessed with regard to there ability to predict impaired fracture healing at an early stage, nevertheless the conclusions of some studies are not consensual. In this article the authors have revised the potential of BTMs as early predictors of prognosis in adult patients presenting traumatic bone fractures but who did not suffer from osteopenia or postmenopausal osteoporosis. The available information from the different studies performed in this field was systematized in order to highlight the most promising BTMs for the assessment of fracture healing outcome.

Serum total and bone alkaline phosphatase levels and their correlation with serum minerals over the lifespan of sheep.

This study aimed to assess serum total alkaline phosphatase (ALP) and its bone isoform (BALP) levels during the ageing and in different physiologic states of sheep, in order to expand the knowledge about the variation of these biomarkers over the sheep lifespan. Ninety female sheep were divided into nine groups of various ages and physiological states (dry, lactation and pregnancy). Serum ALP, BALP and mineral levels were determined by commercial immunoassay, molecular absorbance spectrophotometry and chemical luminescence for BALP determination. Serum ALP and BALP decreased as sheep aged, and no statistically significant differences were obtained between ewes in different physiologic states. The continuous decline of serum BALP concentration along the sheep lifespan, namely in mature and old sheep, is a sign of decreasing bone turnover associated with ageing. Serum calcium concentrations increased slightly until 2 years of age and then showed a tenuous but statistically significant decrease in mature sheep, while serum phosphorus maintained an uninterrupted decrease as sheep matured. The knowledge of serum values of bone biomarkers throughout the sheep lifespan may be useful in preclinical orthopaedic research studies and for animal science studies using sheep.

Comparative Efficacy of iPACK vs Popliteal Sciatic Nerve Block for Pain Management Following Total Knee Arthroplasty: A Retrospective Analysis.

Introduction Total knee arthroplasty (TKA) is associated with severe acute postoperative pain. The use of tourniquets and drains (T/D) is common in TKA but may have an influence on postoperative pain and muscular strength. The infiltration of local anesthetic between the popliteal artery and capsule of the knee (iPACK block) is a motor-sparing block that provides analgesia to the posterior aspect of the knee. However, evidence regarding its efficacy is scarce. This study aims to assess the effectiveness of iPACK block and the impact of T/D use on pain and muscular strength after TKA. Material and methods A retrospective study was carried out including patients who underwent TKA from January 2020 to April 2023. Patients were allocated into groups according to the peripheral nerve block performed and T/D use. Results We included 415 patients in this study. No differences were found in pain at rest or the need for rescue analgesia between patients who received an iPACK block or sciatic nerve block (SNB) with T/D applied. Patients who received a SNB reported lower pain scores on movement (p = 0.019), but with a higher prevalence of motor block (p < 0.001). Patients who underwent surgery without using T/D reported lower pain scores on movement (p = 0.021) and reduced need for rescue analgesia (p = 0.041). Conclusion These findings indicate that iPACK block can facilitate early mobilization after TKA without significant impact on postoperative muscle strength. Furthermore, the use of a T/D may be a source of postoperative pain that could compromise early rehabilitation.

When the Conception of Symmetry Deceives Us: A Case Report on the Perioperative Diagnosis of Subclavian Artery Stenosis.

Subclavian artery stenosis is a relatively uncommon manifestation of peripheral artery disease with significant morbidity. We describe a case of left subclavian artery stenosis that became apparent in the intraoperative setting, in a patient who presented for emergency surgery after a fall and subsequent left femoral neck fracture. Initial non-invasive blood pressure readings on the left upper extremity were in the normal range but after placement of the arterial line on the right upper extremity, the discrepancy was evident pointing towards a structural abnormality as the causative factor. CT angiography was found to confirm the diagnosis of left subclavian artery stenosis of an atherosclerotic nature. A thorough physical examination can point to the presence of subclavian stenosis through pulse amplitude and blood pressure asymmetries. This study highlights the importance of obtaining blood pressure readings ideally on all four limbs to avoid misinterpretation of blood pressure values that could ultimately lead to the use of unnecessary and inadequate interventions and possible complications in the perioperative setting.

Anesthetic Management of a Patient With Prinzmetal Angina.

Prinzmetal angina (PA) is characterized by the development of reversible vasoconstriction of the coronary arteries, transient ischemic electrocardiographic changes in the ST segment, chest pain at rest, and prompt response to nitrates. Spasms of the coronary arteries can be precipitated during the perioperative period by an imbalance of vasodilator and vasoconstrictor factors of smooth muscle cells, which can lead to myocardial ischemia, cardiac arrhythmias, and death. Nevertheless, this is a relatively unrecognized topic, and literature is scarce about it. We present a case report detailing the successful anesthetic management of a patient diagnosed with PA and a documented nitrate allergy, who underwent bilateral ureterorenoscopy.

Clavipectoral Fascia Plane Block Combined With Superficial Cervical Plexus Block for the Removal of Osteosynthesis Material From Clavicle Fracture.

Clavipectoral fascial plane block combined with superficial cervical plexus block has been used as an anesthetic and analgesic technique in mid-clavicle fracture surgeries. The authors describe two cases in which patients underwent extraction of osteosynthesis material from the clavicle, using clavipectoral fascial plane block combined with superficial cervical plexus block as an anesthetic and analgesic technique in the postoperative period. The mentioned block presented itself as an easy-to-perform technique, apparently safe and effective, allowing to obtain satisfactory results.

Polypropylene Modified with Ag-Based Semiconductors as a Potential Material against SARS-CoV-2 and Other Pathogens.

The worldwide outbreak of the coronavirus pandemic (COVID-19) and other emerging infections are difficult and sometimes impossible to treat, making them one of the major public health problems of our time. It is noteworthy that Ag-based semiconductors can help orchestrate several strategies to fight this serious societal issue. In this work, we present the synthesis of α-Ag2WO4, ß-Ag2MoO4, and Ag2CrO4 and their immobilization in polypropylene in the amounts of 0.5, 1.0, and 3.0 wt %, respectively. The antimicrobial activity of the composites was investigated against the Gram-negative bacterium Escherichia coli, the Gram-positive bacterium Staphylococcus aureus, and the fungus Candida albicans. The best antimicrobial efficiency was achieved by the composite with α-Ag2WO4, which completely eliminated the microorganisms in up to 4 h of exposure. The composites were also tested for the inhibition of SARS-CoV-2 virus, showing antiviral efficiency higher than 98% in just 10 min. Additionally, we evaluated the stability of the antimicrobial activity, resulting in constant inhibition, even after material aging. The antimicrobial activity of the compounds was attributed to the production of reactive oxygen species by the semiconductors, which can induce high local oxidative stress, causing the death of these microorganisms.

Antigenotoxic potential of the fermentation broth produced by Paenibacillus polymyxa RNC-D in vitro.

Aim: Evaluate the chemopreventive potential of the extract from P. polymyxa RNC-D. Methods: Concentrations of P. polymyxa RNC-D extract were tested in HepG2/C3A cells to assess their genotoxic (comet assay), mutagenic (micronucleus test) and antigenotoxic potential (comet assay) in vitro. Results: 400 and 40 µg/ml concentrations induced DNA lesions, whereas the 4 µg/ml induced a desmutagenic effect. Complementary tests indicated that the extract minimized the formation of reactive oxygen species induced by methyl methanesulfonate and normalized the loss of membrane potential. The quantification of cytokines indicated that TNF-α was immunostimulated by the extract. However, when administered in conjunction with the methyl methanesulfonate, the extract blocked the TNF-α release. Conclusion: The fermentation broth from P. polymyxa RNC-D showed an antigenotoxic effect, and thus the potential to be used as chemopreventive compound.

Bioactive Ag3PO4/Polypropylene Composites for Inactivation of SARS-CoV-2 and Other Important Public Health Pathogens.

The current unprecedented coronavirus pandemic (COVID-19) is increasingly demanding advanced materials and new technologies to protect us and inactivate SARS-CoV-2. In this research work, we report the manufacture of Ag3PO4 (AP)/polypropylene (PP) composites using a simple method and also reveal their long-term anti-SARS-CoV-2 activity. This composite shows superior antibacterial (against Staphylococcus aureus and Escherichia coli) and antifungal activity (against Candida albicans), thus having potential for a variety of technological applications. The as-manufactured materials were characterized by XRD, Raman spectroscopy, FTIR spectroscopy, AFM, UV-vis spectroscopy, rheology, SEM, and contact angle to confirm their structural integrity. Based on the results of first-principles calculations at the density functional level, a plausible reaction mechanism for the initial events associated with the generation of both hydroxyl radical •OH and superoxide radical anion •O2- in the most reactive (110) surface of AP was proposed. AP/PP composites proved to be an attractive avenue to provide human beings with a broad spectrum of biocide activity.

Cytotoxic Effects and Production of Cytokines Induced by the Endophytic Paenibacillus polymyxa RNC-D In Vitro.

BACKGROUND: Prominent among all the organisms that have a potential value for the production of new medicines, are endophytes, fungi and bacteria that live inside plants without harming them. In this study, a total lyophilized extract (TLE) of Paenibacillus polymyxa RNC-D was used. The P. polymyxa lineages are known for their capacity to segregate a large number of extracellular enzymes and bioactive substances. METHODS: The TLE of Paenibacillus polymyxa RNC-D was tested in cell viability assays for cytotoxicity and cytokine production in BALB/3T3 and J774A.1 cell lineages. RESULTS: A 50% mortality rate of fibroblasts (BALB/3T3) was observed in the 1.171±0.161 mg/mL and 0.956±0.112 mg/mL doses after 48 and 72 hours, respectively, as well as a 50% mortality rate of macrophage cells (J774A.1) in the 0.994±0.170 mg/mL and 0.945±0.280 mg/mL doses after 48 and 72 hours, respectively. The ≈1 mg/mL concentration significantly affected the kinetic of growth in all the measured periods. The extract induced apoptosis and necrosis 24 hours after the ≈1 mg/mL concentration in both tested lineages. The treatment with the ≈1 mg/mL concentration led to the production of TNF-α and IFN-γ cytokines in 24 hours. IL-12 and IL-10 began to be detected as a result of the treatment with 0.1 mg/mL. However, with the 0.5 mg/mL dose in 24 hours, a significant reduction in IL-10 was observed. CONCLUSION: Our data suggest that the TLE of P. polymyxa RNC-D modulated the production of cytokines with different patterns of immune response in a dose-dependent way.

Evaluation of bone turnover markers and serum minerals variations for predicting fracture healing versus non-union processes in adult sheep as a model for orthopedic research.

Bone turnover markers (BTMs) have been considered as an auxiliary method of following the fracture healing process and for early prediction of impaired bone healing. A better understanding of the potential of BTMs in this application could allow for earlier interventions and improved patient care. The aim of this study with a large animal experimental model was to assess the variation of bone formation markers - namely the total alkaline phosphatase (ALP) and its bone-specific isoform (BALP), serum concentration of intact osteocalcin (OC), N-terminal propeptide type III procollagen (PIIINP) and of bone resorption markers - namely tartrate resistant acid phosphatase (TRAP) and deoxypyridinoline crosslink (DPD) during the first stages of a normal fracture healing process and of a segmental critical size defect (CSD), which progresses to a non-union process. Thirty healthy female sheep (Portuguese Churra-da-Terra-Quente breed), approximately 4-years-old, were enrolled in this study. Jugular venous blood samples were collected pre-operatively and at 1, 2, 3, 4, 6, 8, 10 and 12 post-operative weeks. The animals of the CSD group showed significant lower serum levels of BALP, OC and significant higher serum PIIINP levels at early stages of the fracture healing process, compared with animals that progressed in a normal fracture healing process. Serum BALP, OC and PIIINP levels could be useful as non-invasive auxiliary tools with other complementary methods for predicting the outcome of traumatic bone fractures.

Short-term variability in biomarkers of bone metabolism in sheep.

Changes in bone remodeling during pathological states and during their treatment can be assessed noninvasively by measuring biomarkers of bone metabolism. Their application is limited, however, by the potential biological variability in the levels of these biomarkers over time. To determine the short-term variability in biomarkers of bone metabolism in adult sheep, the authors measured serum levels of alkaline phosphatase (ALP), bone-specific alkaline phosphatase (BALP), osteocalcin (OC), N-terminal propeptide of type-III procollagen (PIIINP), deoxypyridinoline (DPD), tartrate-resistant acid phosphatase (TRAP), calcium and phosphorus intermittently over a 12-week period. There were significant differences in mean ALP activity and in phosphorus concentrations over time, but all other biomarkers showed no significant short-term variability. The results suggest that biomarkers of bone metabolism in sheep, especially the bone resorption marker DPD and the bone formation marker BALP, can be used reliably to detect changes in bone cellular activity.

Bone turnover markers in sheep and goat: A review of the scientific literature

ABSTRACT Bone turnover markers (BTMs) are product of bone cell activity and are generally divided in bone formation and bone resorption markers. The purpose of this review was to structure the available information on the use of BTMs in studies on small ruminants, especially for monitoring their variations related to diet, exercise, gestation and metabolic lactation state, circadian and seasonal variations, and also during skeletal growth. Pre-clinical and translational studies using BTMs with sheep and goats as animal models in orthopaedic research studies to help in the evaluation of the fracture healing process and osteoporosis research are also described in this review. The available information from the reviewed studies was systematically organized in order to highlight the most promising BTMs in small ruminant research, as well as provide a wide view of the use of sheep and goat as animal models in orthopaedic research, type of markers and commercial assay kits with cross-reactivity in sheep and goat, method of sample and storage of serum and urine for bone turnover markers determination and the usefulness and limitations of bone turnover markers in the different studies, therefore an effective tool for researchers that seek answers to different questions while using BTMs in small ruminants.