Biological hypoxia in pre-transplant human pancreatic islets induces transplant failure in diabetic mice.

Evaluating the quality of isolated human islets before transplantation is crucial for predicting the success in treating Type 1 diabetes. The current gold standard involves time-intensive in vivo transplantation into diabetic immunodeficient mice. Given the susceptibility of isolated islets to hypoxia, we hypothesized that hypoxia present in islets before transplantation could indicate compromised islet quality, potentially leading to unfavorable outcomes. To test this hypothesis, we analyzed expression of 39 hypoxia-related genes in human islets from 85 deceased donors. We correlated gene expression profiles with transplantation outcomes in 327 diabetic mice, each receiving 1200 islet equivalents grafted into the kidney capsule. Transplantation outcome was post-transplant glycemic control based on area under the curve of blood glucose over 4 weeks. In linear regression analysis, DDIT4 (R = 0.4971, P < 0.0001), SLC2A8 (R = 0.3531, P = 0.0009) and HK1 (R = 0.3444, P = 0.0012) had the highest correlation with transplantation outcome. A multiple regression model of 11 genes increased the correlation (R = 0.6117, P < 0.0001). We conclude that assessing pre-transplant hypoxia in human islets via gene expression analysis is a rapid, viable alternative to conventional in vivo assessments. This approach also underscores the importance of mitigating pre-transplant hypoxia in isolated islets to improve the success rate of islet transplantation.

Smoking is not closely related to revision for periprosthetic joint infection after primary total knee and hip arthroplasty.

BACKGROUND: The influence of smoking on the risk of periprosthetic joint infection (PJI) remains unclear. The objective was to explore the impact of smoking on PJI after primary total knee (TKA) and hip (THA) arthroplasty. HYPOTHESIS: Current smoking patients should have an increased risk of PIJ compared with nonsmoking patients. PATIENTS AND METHODS: A prospective registry-based observational cohort study was performed. A total of 4591 patients who underwent primary TKA (3076 patients) or THA (1515) were included. According to the smoking status at the time of arthroplasty, patients were classified as nonsmokers (3031 patients), ex-smokers (688), and smokers (872). Multivariate analysis included smoking status, age, gender, education level, body mass index, American Society of Anesthesiologists class, diagnosis (osteoarthritis, rheumatism), diabetes, chronic obstructive pulmonary disease, perioperative blood transfusion, site of arthroplasty (knee, hip), length of operation, and length of stay. RESULTS: There were PJI after 59 (1.9%) TKA and 27 (1.8%) THA (p=0.840). There were PJI in 47 (1.6%) nonsmokers, 12 (1.7%) ex-smokers, and 17 (1.9%) smokers (p=0.413). There were wound complications (delayed wound healing and superficial wound infection) in 34 (0.7%) nonsmokers, 9 (1.3%) in ex-smokers, and 17 (1.9%) in smokers (p=0.045). In multivariate analysis, only the female gender was a significant predictor of PJI (OR 1.3, 95% CI 1.1-2.4 [p=0.039]). Specifically, the categories of ex-smokers (OR 0.8, 95% CI 0.2-1.7 [p=0.241]) and smokers (OR 1.1, 95% CI 0.6-1.5 [p=0.052]) were not significant predictors. The 4-year arthroplasty survival with PJI as the endpoint was 99.1% (95% CI: 99.0-99.7) for nonsmokers, 99.0% (95% CI: 98.8-99.2) for ex-smokers, and 98.7% (95% CI: 98.2-99.0) for smokers was not significantly different between smoking status groups (p=0.318). DISCUSSION: Smoking was not identified as a significant predictor for PJI following primary TKA or THA. LEVEL OF EVIDENCE: III, prospective cohort study.

The relative importance of salinization in lowland stream zooplankton: Implications of the ecosystem nutrient status.

Salinization of aquatic systems is predicted to increase due to climate and land use changes. Nevertheless, community responses may be different according to the ecosystem characteristics and contextual scenarios. Small flowing waters are particularly vulnerable to salinization, which may impact on the biodiversity and ecosystem processes, but this remains unclear. We conducted a study in 42 lowland streams characterized by overall high nutrient levels along a salinity gradient between 2 and 160 g L-1 to analyze changes in zooplankton structural and functional metrics, and the grazing effects of zooplankton on phytoplankton affecting the energy transfer. Generalized additive models revealed that the analyzed metrics were relatively influenced by salinity, with factors related to trophic conditions playing an important role as well. Total abundance and biomass decreased along the salinity gradient while increasing at intermediate soluble reactive phosphorous concentrations (SRP) in the former and with a linear increase in the SRP in the latter case. Taxonomic richness decreased with salinity and dissolved inorganic nitrogen, with species replacement toward saline-tolerant ones according to the compositional and optimums analyses. In opposite, functional richness did not display any specific trend within the environmental gradients. This explains why zooplankton compositional changes were not reflected into shifts in the grazing pattern on phytoplankton, which was in turn driven by SRP and dissolved oxygen concentrations. Further research is a critical requirement in these poorly studied ecosystems for planning mitigation actions to the co-occurrence of eutrophication and salinization in a fast changing world.

A robust knock-in approach using a minimal promoter and a minicircle.

Knock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated that minicircles serve as an efficient donor DNA in zebrafish, significantly enhancing KI events compared to plasmids containing bacterial backbones. In an attempt to generate a KI reporter for scn8ab, we precisely integrated a fluorescence gene at the start codon. However, the seamlessly integrated reporter was unable to direct expression that recapitulates endogenous scn8ab expression. To overcome this obstacle, we introduced the hsp70 minimal promoter to provide an ectopic transcription initiation site and succeeded in establishing stable KI transgenic reporters for scn8ab. This strategy also created a fgf20b KI reporter line with a high success rate. Furthermore, our data revealed that an unexpectedly edited genome can inappropriately influence the integrated reporter gene expression, highlighting the importance of selecting a proper KI line. Overall, our approach utilizing a minicircle and an ectopic promoter establishes a robust and efficient strategy for KI generation, expanding our capacity to create KI animals.

A robust knock-in approach using a minimal promoter and a minicircle.

Knock-in reporter (KI) animals are essential tools in biomedical research to study gene expression impacting diverse biological events. While CRISPR/Cas9-mediated genome editing allows for the successful generation of KI animals, several factors should be considered, such as low expression of the target gene, prevention of bacterial DNA integration, and in-frame editing. To circumvent these challenges, we developed a new strategy that utilizes minicircle technology and introduces a minimal promoter. We demonstrated that minicircles serve as an efficient donor DNA in zebrafish, significantly enhancing KI events compared to plasmids containing bacterial backbones. In an attempt to generate a KI reporter for scn8ab, we precisely integrated a fluorescence gene at the start codon. However, the seamlessly integrated reporter was unable to direct expression that recapitulates endogenous scn8ab expression. To overcome this obstacle, we introduced the hsp70 minimal promoter to provide an ectopic transcription initiation site and succeeded in establishing stable KI transgenic reporters for scn8ab. This strategy also created a fgf20b KI reporter line with a high success rate. Furthermore, our data revealed that an unexpectedly edited genome can inappropriately influence the integrated reporter gene expression, highlighting the importance of selecting a proper KI line. Overall, our approach utilizing a minicircle and an ectopic promoter establishes a robust and efficient strategy for KI generation, expanding our capacity to create KI animals.

Beneficios del consumo de dieta mediterránea en individuos con elevado riesgo cardiovascular: un patrón que debe convertirse en el común denominador

Las enfermedades cardiovasculares siguen siendo la principal causa de morbilidad y mortalidad en el mundo. Y aunque existen herramientas terapéuticas y farmacológicas que ayudan a controlar la incidencia de desenlaces cardiovasculares con la reducción del riesgo cardiovascular, las inequidades en el acceso oportuno a servicios de salud y de determinantes sociales de la salud impiden que toda la población con esta necesidad pueda recibir esta ayuda terapéutica. Es así, como se da prioridad a las intervenciones no farmacológicas, dentro de las que se destaca, la dieta, la cual ha demostrado influir significativamente en la salud cardiovascular y en la expectativa de vida del ser humano. Particularmente, la dieta mediterránea, basada esencialmente en el consumo de frutas, vegetales, granos, legumbres, aceite de oliva, especias, seguido de proteína derivada de pescado y comida de mar, ha demostrado proveer cierto grado de protección frente a enfermedades cardiometabólicas. Sin embargo, se desconoce con certeza cuál es el impacto en la población con elevado riesgo cardiovascular y, por ende, enfermedad cardiovascular establecida. Considerando la evolución de la evidencia en los patrones dietarios, así como la necesidad de conocer qué tanto impactan las dietas viables en nuestro medio, sobre todo en aquellos con mayor riesgo de morbilidad y mortalidad por enfermedad cardiovascular, el objetivo de esta revisión consiste en exponer evidencia actualizada sobre los beneficios de la dieta mediterránea en individuos con elevado riesgo cardiovascular. Para esto, se realizó una búsqueda bibliográfica en las bases de datos PubMed, Science Direct y Scielo.

Cardiovascular diseases continue to be the leading cause of morbidity and mortality worldwide. Although there are therapeutic and pharmacological tools that help control the incidence of cardiovascular outcomes by reducing cardiovascular risk, inequities in timely access to healthcare services and social determinants of health prevent the entire population in need from receiving this therapeutic assistance. Thus, non-pharmacological interventions are prioritized, among which the diet stands out, as it has been shown to significantly influence cardiovascular health and life expectancy. Particularly, the Mediterranean diet, primarily based on the consumption of fruits, vegetables, grains, legumes, olive oil, spices, followed by fish and seafood-derived protein, has demonstrated a certain degree of protection against cardiometabolic diseases. However, the impact on a population with high cardiovascular risk and established cardiovascular disease remains uncertain. Considering the evolving evidence on dietary patterns and the need to understand how viable diets in our context affect individuals with a higher risk of morbidity and mortality from cardiovascular disease, the objective of this review is to present updated evidence on the benefits of the Mediterranean diet in individuals with high cardiovascular risk. For this purpose, a literature search was performed in the PubMed, Science Direct, and Scielo databases.

Phytochemical characterization and antifungal potential of leaf extracts of Mosiera bullata.

The investigation of natural alternatives to conventional fungicides is of imminent need. Mosiera bullata (Britton & P. Wilson) Bisse is a Cuban endemic plant species belonging to the Myrtaceae family. The objective of the present study was to perform a bioassay-guided fractionation to explore the potential of extracts and fractions from M. bullata leaves against a panel of fungal plant pathogens. The M. bullata total extract was confirmed to have good antifungal activity against R. oryzae (IC50 = 4.86 µg/mL) and moderate activity against F. oxysporum (IC50 = 352.40 µg/mL) and F. solani (IC50 = 427.38 µg/mL) and fungicidal effect against R. oryzae. Five compounds belonging to the class of phloroglucinol dimers were tentatively characterized by UHPLC-HRMS and reported for the first time in M. bullata and the genus Mosiera. These results suggest the potential of M. bullata total extract as a natural antifungal product for the control of diseases in agriculture.

Exoprotease exploitation and social cheating in a Pseudomonas aeruginosa environmental lysogenic strain with a noncanonical quorum sensing system.

Social cheating is the exploitation of public goods that are costly metabolites, like exoproteases. Exoprotease exploitation in Pseudomonas aeruginosa has been studied in reference strains. Experimental evolution with reference strains during continuous growth in casein has demonstrated that nonexoprotease producers that are lasR mutants are selected while they behave as social cheaters. However, noncanonical quorum-sensing systems exist in P. aeruginosa strains, which are diverse. In this work, the exploitation of exoproteases in the environmental strain ID4365 was evaluated; ID4365 has a nonsense mutation that precludes expression of LasR. ID4365 produces exoproteases under the control of RhlR, and harbors an inducible prophage. As expected, rhlR mutants of ID4365 behave as social cheaters, and exoprotease-deficient individuals accumulate upon continuous growth in casein. Moreover, in all continuous cultures, population collapses occur. However, this also sometimes happens before cheaters dominate. Interestingly, during growth in casein, ID4565's native prophage is induced, suggesting that the metabolic costs imposed by social cheating may increase its induction, promoting population collapses. Accordingly, lysogenization of the PAO1 lasR mutant with this prophage accelerated its collapse. These findings highlight the influence of temperate phages in social cheating.

Myriad applications of bacteriophages beyond phage therapy.

Bacteriophages are the most abundant biological entity on the planet, having pivotal roles in bacterial ecology, animal and plant health, and in the biogeochemical cycles. Although, in principle, phages are simple entities that replicate at the expense of their bacterial hosts, due the importance of bacteria in all aspects of nature, they have the potential to influence and modify diverse processes, either in subtle or profound ways. Traditionally, the main application of bacteriophages is phage therapy, which is their utilization to combat and help to clear bacterial infections, from enteric diseases, to skin infections, chronic infections, sepsis, etc. Nevertheless, phages can also be potentially used for several other tasks, including food preservation, disinfection of surfaces, treatment of several dysbioses, and modulation of microbiomes. Phages may also be used as tools for the treatment of non-bacterial infections and pest control in agriculture; moreover, they can be used to decrease bacterial virulence and antibiotic resistance and even to combat global warming. In this review manuscript we discuss these possible applications and promote their implementation.

Epigenetic and transcriptomic characterization reveals progression markers and essential pathways in clear cell renal cell carcinoma.

Identifying tumor-cell-specific markers and elucidating their epigenetic regulation and spatial heterogeneity provides mechanistic insights into cancer etiology. Here, we perform snRNA-seq and snATAC-seq in 34 and 28 human clear cell renal cell carcinoma (ccRCC) specimens, respectively, with matched bulk proteogenomics data. By identifying 20 tumor-specific markers through a multi-omics tiered approach, we reveal an association between higher ceruloplasmin (CP) expression and reduced survival. CP knockdown, combined with spatial transcriptomics, suggests a role for CP in regulating hyalinized stroma and tumor-stroma interactions in ccRCC. Intratumoral heterogeneity analysis portrays tumor cell-intrinsic inflammation and epithelial-mesenchymal transition (EMT) as two distinguishing features of tumor subpopulations. Finally, BAP1 mutations are associated with widespread reduction of chromatin accessibility, while PBRM1 mutations generally increase accessibility, with the former affecting five times more accessible peaks than the latter. These integrated analyses reveal the cellular architecture of ccRCC, providing insights into key markers and pathways in ccRCC tumorigenesis.

Work-from-home is here to stay: Call for flexibility in post-pandemic work policies.

In early 2020, the Covid-19 pandemic forced employees in tech companies worldwide to abruptly transition from working in offices to working from their homes. During two years of predominantly working from home, employees and managers alike formed expectations about what post-pandemic working life should look like. Many companies are experimenting with new work policies that balance employee- and manager expectations regarding where, when and how work should be done in the future. In this article, we gather experiences of the new trend of remote working based on the synthesis of 22 company-internal surveys of employee preferences for WFH, and 26 post-pandemic work policies from 17 companies and their sites, covering 12 countries in total. Our results are threefold. First, through the new work policies, all companies formally give employees more flexibility regarding working time and location. Second, there is a great variation in how much flexibility the companies are willing to yield to the employees. The paper details the different formulations that companies adopted to document the extent of permitted WFH, exceptions, relocation permits and the authorisation procedures. Third, we document a change in the psychological contract between employees and managers, where the option of working from home is converted from an exclusive perk that managers could choose to give to the few, to a core privilege that all employees feel they are entitled to. Finally, there are indications that as the companies learn and solicit feedback regarding the efficiency of the chosen strategies, we will see further developments and changes in the work policies concerning how much flexibility to work whenever and from wherever they grant. Through these findings, the paper contributes to a growing literature about the new trends emerging from the pandemic in tech companies and spells out practical implications onwards.

Inhibition of Bacterial Adhesion and Biofilm Formation by Seed-Derived Ethanol Extracts from Persea americana Mill.

The increase in antibiotic resistance demands innovative strategies to combat microorganisms. The current study evaluated the antibacterial and antivirulence effects of ethanol extracts from Persea americana seeds obtained by the Soxhlet (SE) and maceration (MaE) methods. The UHPLC-DAD-QTOF analysis showed mainly the presence of polyphenols and neolignan. Ethanol extracts were not cytotoxic to mammalian cells (CC50 > 500 µg/mL) and displayed a moderate antibacterial activity against Pseudomonas aeruginosa (IC50 = 87 and 187 µg/mL) and Staphylococcus aureus (IC50 = 144 and 159 µg/mL). Interestingly, no antibacterial activity was found against Escherichia coli. SE and MaE extracts were also able to significantly reduce the bacterial adhesion to A549 lung epithelial cells. Additionally, both extracts inhibited the biofilm growth at 24 h and facilitated the release of internal cell components in P. aeruginosa, which might be associated with cell membrane destabilization. Real-time PCR and agarose electrophoresis gel analysis indicated that avocado seed ethanol extracts (64 µg/mL) downregulated virulence-related factors such as mexT and lasA genes. Our results support the potential of bioproducts from P. americana seeds as anti-adhesive and anti-biofilm agents.

Electrosprayed Agar Nanocapsules as Edible Carriers of Bioactive Compounds.

Electrosprayed agar nanocapsules were developed using an acetic acid solution as solvent. The role of solution properties (viscosity, surface tension, and conductivity) in the formation of agar particles was assessed, together with the effect of both agar and acetic acid concentrations on the size and morphology of the resulting particles. Agar solutions with a concentration below 10% w/v were not suitable for electrospraying. Furthermore, the agar-acetic acid ratio was also critical for the formation of agar nanostructures (with an optimum ratio of 1:2). A decrease in particle size was also observed when decreasing agar concentration, with particle diameter values ranging between 50 and 400 nm. Moreover, the suitability of the electrosprayed agar nanocapsules as carriers for a model bioactive compound, chlorophyllin sodium copper salt (CHL), was also evaluated. The release profile of encapsulated CHL, with an estimated encapsulation efficiency of around 40%, was carried out in food simulants with different hydrophilicity (10% v/v and 50% v/v ethanol). While the release of the bioactive was negligible in the hydrophilic food simulant, an initial burst release followed by a slower sustained release was observed when the capsules were immersed in 50% ethanol solution. The results open up a broad range of possibilities that deserve further exploration related to the use of these edible polysaccharide-based nanocapsules.

Voltage-gated sodium channel scn8a is required for innervation and regeneration of amputated adult zebrafish fins.

Teleost fishes and urodele amphibians can regenerate amputated appendages, whereas this ability is restricted to digit tips in adult mammals. One key component of appendage regeneration is reinnervation of the wound area. However, how innervation is regulated in injured appendages of adult vertebrates has seen limited research attention. From a forward genetics screen for temperature-sensitive defects in zebrafish fin regeneration, we identified a mutation that disrupted regeneration while also inducing paralysis at the restrictive temperature. Genetic mapping and complementation tests identify a mutation in the major neuronal voltage-gated sodium channel (VGSC) gene scn8ab. Conditional disruption of scn8ab impairs early regenerative events, including blastema formation, but does not affect morphogenesis of established regenerates. Whereas scn8ab mutations reduced neural activity as expected, they also disrupted axon regrowth and patterning in fin regenerates, resulting in hypoinnervation. Our findings indicate that the activity of VGSCs plays a proregenerative role by promoting innervation of appendage stumps.

Functional tremor developing after successful MRI-guided focused ultrasound thalamotomy for essential tremor.

OBJECTIVE: To describe a case of functional tremor occurring after a successful MR-guided focused ultrasound thalamotomy (MRgFUS) for essential tremor. METHODS: A 71-year-old right-handed man with essential tremor was referred to us for consideration of deep brain stimulation surgery for worsening bilateral upper limb tremor after a successful left MRgFUS for essential tremor. RESULTS: On clinical exam, signs compatible with a functional tremor were noted, including entertainability and suppressibility. Electrophysiological studies were consistent with essential tremor and superimposed tremor fulfilling the laboratory-supported criteria for functional tremor. DISCUSSION: We describe the first reported case of a functional movement disorder occurring after successful MRgFUS procedure for essential tremor. Recognising this entity and its development after such therapeutic interventions is essential to avoid further unnecessary invasive therapies.

A validated outcome categorization of the knee society score for total knee arthroplasty.

PURPOSE: To determine cutoff values for the Knee Society Scores (KSS) indicative of a categorical scale of medium-term outcomes. METHODS: One hundred and fifty-five patients who underwent primary cruciate-retaining TKA with a patellar button for osteoarthritis at a single-centre were assessed prospectively by the KSS and short-form Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) simultaneously at the 3-year follow-up. A validated categorization of the WOMAC score was used as a standard. The area under the curve (AUC) of receiver-operating characteristic (ROC) was used to assess the discriminative analysis accuracy of the, and the Youden index estimated the optimal cutoff point. RESULTS: For the KSS-knee score, the cutoff for an excellent outcome was 90.3 (AUC 0.75, 95% CI 0.71-0.78), 76.6 (AUC 76.6, 95% CI 0.70-076) for good, 64.8 (AUC 0.76, 95% CI 0.72-0.79) for fair, and < 64.8 (AUC 0.69, 95% CI 0.67-0.73) for poor. For the KSS-function score, the cutoff values were 85.2 (AUC 0.71, 95% CI 0.69-0.75), 73.1 (AUC 0.72, 95% CI, 0.70-0.76), 55.7 (AUC 0.70, 95% CI 0.71-0.74), and < 55.7 (AUC 0.68, 95% CI 0.66-0.72), respectively. CONCLUSION: A KSS-knee score ≥ of 90 was considered an excellent outcome, 77 good, 65 fair, and < 65 poor. For the KSS-function, those values are 85, 73, 56 and < 56, respectively. The treatment outcome's judgement may be clearer for the surgeon concerning a particular patient when using cutoff values for the scoring system employed, such as those determined in the present study. LEVEL OF EVIDENCE: II.

Combined use of monensin and virginiamycin to improve rumen and liver health and performance of feedlot-finished steers.

Monensin and virginiamycin are included in beef cattle finishing diets as prophylaxis to minimize the incidence of ruminal acidosis and liver abscesses. Due to different and probably complementary modes of action, this study aimed to determine the effects of a combination of monensin and virginiamycin, both included in the diet at recommended doses, on ruminal health, the occurrence of liver abscesses, and growth performance of feedlot-finished cattle. One hundred and forty-four steers (6 animals/pen) were fed 1 of 3 corn-based finishing diets containing 30 mg of monensin (MN), 25 mg of virginiamycin (VM), or 30 and 25 mg of monensin and virginiamycin (MN + VM), respectively, per kilogram of dry matter. Ruminal pH probes were inserted into two animals per pen and set to record pH every 10 min. On d 100, animals were slaughtered, and rumens and livers were recovered, on which occurrence and degree of ruminal damage, prevalence and number of liver abscesses, and liver scores (A-: livers with no more than two small abscesses; A+: livers with at least one large abscess or more than four medium abscesses; A: any other abscessed liver) were determined. Simultaneous inclusion of monensin and virginiamycin resulted in a 4.3% decrease (P < 0.04) in dry matter intake (DMI; 8.8, 9.2, and 9.2 ± 0.19 kg/d for MN + VM, MN, and VM-fed animals, respectively) and similar (P > 0.13) average daily body weight gain (ADG; 1.49 ± 0.021 kg/d) and hot carcass weight (HCW; 269 ± 1.7 kg), compared with feeding diets containing one additive or the other. Therefore, in terms of ADG, a 9.4% improvement (P < 0.01) in feed efficiency was observed in MN + VM-fed animals. Backfat thickness (5.6 ± 0.08 mm) and ribeye area (69.9 ± 0.53 cm2) remained unaffected (P ≥ 0.74), as well as the minimum (4.98 ± 0.047), mean (6.11 ± 0.037), and maximum ruminal pH (7.23 ± 0.033) values and the time (125 ± 22.3 min/d), area (57.67 ± 12.383 pH × h), and episodes (22 ± 3.8 bouts) of pH below 5.6 (P ≥ 0.12). Overall, prevalence (24 ± 3.4%) and the number of liver abscesses (1.6 ± 0.14 abscesses/abscessed liver), liver scores (20 ± 3.1% of A- and 4 ± 1.8% of A livers), and prevalence (67 ± 3.5%) and degree of damage to the ruminal epithelium (2.5 ± 0.22% affected surface) were similar (P ≥ 0.18) across treatments; however, the occurrence of ruminal lesions tended (P ≤ 0.07) to be associated with that of liver abscesses and reduced ADG when feeding monensin alone.

Antifungal Activity of Extracts, Fractions, and Constituents from Coccoloba cowellii Leaves.

Coccoloba cowellii Britton (Polygonaceae, order Caryophyllales) is an endemic and critically endangered plant species that only grows in the municipality of Camagüey, a province of Cuba. A preliminary investigation of its total methanolic extract led to the discovery of promising antifungal activity. In this study, a bioassay-guided fractionation allowed the isolation of quercetin and four methoxyflavonoids: 3-O-methylquercetin, myricetin 3,3',4'-trimethyl ether, 6-methoxymyricetin 3,4'-dimethyl ether, and 6-methoxymyricetin 3,3',4'-trimethyl ether. The leaf extract, fractions, and compounds were tested against various fungi and showed strong in vitro antifungal activity against Cryptococcus neoformans and various Candida spp. with no cytotoxicity (CC50 > 64.0 µg/mL) on MRC-5 SV2 cells, determined by a resazurin assay. A Candida albicans SC5314 antibiofilm assay indicated that the antifungal activity of C. cowellii extracts and constituents is mainly targeted to planktonic cells. The total methanolic extract showed higher and broader activity compared with the fractions and mixture of compounds.

Bioactive extracts from persimmon waste: influence of extraction conditions and ripeness.

In this work, a bioactive persimmon extract was produced from discarded fruits. A central composite design was used to evaluate the effect of different extraction parameters and ripeness stages of persimmon fruits on the total phenolic content and antioxidant activity of the resulting extracts. Significantly greater phenolic contents were obtained from immature persimmon (IP) fruits. The optimum IP extract with the conditions set by the experimental design was industrially up-scaled and its composition and functional properties were evaluated and compared with those obtained under lab-scale conditions. Both extracts contained significant protein (>20%) and phenolic contents (∼11-27 mg GA/g dry extract) and displayed significant antiviral activity against murine norovirus and hepatitis A virus. Moreover, the extract showed no toxicity and significantly reduced the fat content and the cellular ageing of Caenorhabditis elegans (C. elegans) without affecting the worm development. These effects were mediated by down-regulation of fat-7, suggesting an anti-lipogenic activity of this extract.