In vitro anti-tumoral and immunomodulatory effects of acidic polysaccharides isolated from the fungus Gloeosoma mirabile.

Polysaccharides from wood-rooting fungi have attracted attention due to their broad pharmacological properties. Herein, we report the antitumor and immunomodulatory activities of acid polysaccharides isolated from fungi Gloeosoma mirabile. The polysaccharide extracts displayed significant antiproliferative activity against cancer cell lines (MCF-7, HCT-116, U-937) in a dose-dependent manner and induction of IL-6 in macrophage RAW 264.7. Furthermore, flow cytometry analysis showed that high polysaccharide concentrations induced apoptosis by 83% in HL-60 cells. Based on gas chromatography-mass spectrometry (GC-MS) and Fourier transform infra-red (FT-IR) spectroscopy studies, acidic polysaccharides from G. mirabile were mainly composed of arabinose, α-D-galactopyranose and methyl ß-D-galactopyranoside.

Antioxidant Responses and Phytochemical Accumulation in Raphanus Species Sprouts through Elicitors and Predictive Models under High Temperature Stress.

Crop production is being impacted by higher temperatures, which can decrease food yield and pose a threat to human nutrition. In the current study, edible and wild radish sprouts were exposed to elevated growth temperatures along with the exogenous application of various elicitors to activate defense mechanisms. Developmental traits, oxidative damage, glucosinolate and anthocyanin content, and antioxidant capacity were evaluated alongside the development of a predictive model. A combination of four elicitors (citric acid, methyl jasmonate-MeJa, chitosan, and K2SO4) and high temperatures were applied. The accumulation of bioactives was significantly enhanced through the application of two elicitors, K2SO4 and methyl jasmonate (MeJa). The combination of high temperature with MeJa prominently activated oxidative mechanisms. Consequently, an artificial neural network was developed to predict the behavior of MeJa and temperature, providing a valuable projection of plant growth responses. This study demonstrates that the use of elicitors and predictive analytics serves as an effective tool to investigate responses and enhance the nutritional value of Raphanus species sprouts under future conditions of increased temperature.

Nanocarrier of α-Tocopheryl Succinate Based on a Copolymer Derivative of (4,7-dichloroquinolin-2-yl)methanol and Its Cytotoxicity against a Breast Cancer Cell Line.

In order to improve the water solubility and, therefore, bioavailability and therapeutic activity of anticancer hydrophobic drug α-tocopherol succinate (α-TOS), in this work, copolymers were synthesized via free radicals from QMES (1-[4,7-dichloroquinolin-2-ylmethyl]-4-methacryloyloxyethyl succinate) and VP (N-vinyl-2-pirrolidone) using different molar ratios, and were used to nanoencapsulate and deliver α-TOS into cancer cells MCF-7. QMES monomer was chosen because the QMES pendant group in the polymer tends to hydrolyze to form free 4,7-dichloro-2-quinolinemethanol (QOH), which also, like α-TOS, exhibit anti-proliferative effects on cancerous cells. From the QMES-VP 30:70 (QMES-30) and 40:60 (QMES-40) copolymers obtained, it was possible to prepare aqueous suspensions of empty nanoparticles (NPs) loaded with α-TOS by nanoprecipitation. The diameter and encapsulation efficiency (%EE) of the QMES-30 NPs loaded with α-TOS were 128.6 nm and 52%; while for the QMES-40 NPs loaded with α-TOS, they were 148.8 nm and 65%. The results of the AlamarBlue assay at 72 h of treatment show that empty QMES-30 NPs (without α-TOS) produced a marked cytotoxic effect on MCF-7 breast cancer cells, corresponding to an IC50 value of 0.043 mg mL-1, and importantly, they did not exhibit cytotoxicity against healthy HUVEC cells. Furthermore, NP-QMES-40 loaded with α-TOS were cytotoxic with an IC50 value of 0.076 mg mL-1, demonstrating a progressive release of α-TOS; however, the latter nanoparticles were also cytotoxic to healthy cells in the range of the assayed concentrations. These results contribute to the search for a new polymeric nanocarrier of QOH, α-TOS or other hydrophobic drugs for the treatment of cancer or others diseases treatable with these drugs.

Comparison of serum creatinine, point-of-care symmetric dimethylarginine and renal imaging with glomerular filtration rate measured by renal scintigraphy in healthy and early chronic kidney diseased cats.

The aim of this study was to evaluate routinely used tests to diagnose cats in early stages of chronic kidney disease (CKD) and to describe a model for evaluating these variables simultaneously. Apparently healthy cats were screened using serum creatinine (sCr), point-of-care symmetric dimethylarginine (POC SDMA), urinalysis, urine protein/creatinine ratio (UPC) and imaging evaluation. Those parameters were compared to glomerular filtration rate (GFR) assessed by renal scintigraphy. Forty-four cats were included and consisted of 14 (31.8%) healthy cats (absence of abnormalities in renal morphology and sCr less than 1.6 mg/dL), 20 (45.5%) cats classified as CKD I (presence of abnormalities in renal morphology and sCr less than 1.6 mg/dL) and ten (22.7%) as CKD II (sCr equal to or greater than 1.6 mg/dL, with or without abnormalities in renal morphology). A large number (40.9%) of apparently healthy cats presented reduction in GFR, which included half of CKD I patients. Point-of-care SDMA was not a good predictor for decreased GFR, nor was it correlated with the variables GFR and sCr. Glomerular filtration rate was significantly lower in CKD I and II groups in comparison with healthy cats, but there was no significant difference between the CKD I and II groups. Multivariate logistic regression model identified three variables that affected the odds of a cat having decreased GFR (< 2.5 mL/min/kg): sCr (OR = 18.3; p = 0.019; CI = 1.6-207.2), and the ultrasonographic findings 'reduced corticomedullary definition' (OR = 19.9; p = 0.022; CI = 1.6-254.0) and 'irregular contour' (OR = 65.6; p = 0.003; CI = 4.2-1038.2). Renal ultrasonography evaluation should always be considered for screening early CKD in apparently healthy cats.

Influence of Constant Rate Infusions of Fentanyl Alone or in Combination With Lidocaine and Ketamine on the Response to Surgery and Postoperative Pain in Isoflurane Anesthetized Dogs Undergoing Unilateral Mastectomy: A Randomized Clinical Trial.

The aim of this study was to compare the effects of constant rate infusions (CRI) of fentanyl alone or combined with lidocaine and ketamine (FLK), on physiological parameters, isoflurane requirements and the number of postoperative analgesic rescues in dogs undergoing unilateral mastectomy. Twenty-two dogs were premedicated with acepromazine 0.02 mg/kg and morphine 0.5 mg/kg and anesthetized with propofol and isoflurane. Dogs were randomly assigned to 1 of 2 groups: Fentanyl group (fentanyl 5 µg/kg loading dose [LD] and 9 µg/kg/h CRI; n = 11); FLK group (fentanyl [same doses]; lidocaine 2 mg/kg LD and 3 mg/kg/h CRI; ketamine 1.0 mg/kg LD and 0.6 mg/kg/h CRI; = 11). Intraoperative evaluations were performed before the start of surgery and administration of the treatments (T0); three minutes after the LD (T1); during incision and tissue divulsion (T2); during closure of the surgical wound (T3). Meloxicam (0.1 mg/kg) was administered at T3. Blood samples were collected for determination of plasma concentrations of fentanyl, lidocaine and ketamine. Pain scores and the number of postoperative analgesic rescues with morphine (0.5 mg/kg) were evaluated for 24 hours postoperatively using the short form of the Glasgow Composite Measure Pain Scale. Compared to T0, significant decreases in heart rate (from 84 ± 28 to 53 ± 16 bpm in the Fentanyl group and from 93 ± 16 to 63 ± 15 bpm in FLK) and mean arterial pressure (from 61 ± 5 to 49 ± 10 mmHg in Fentanyl and from 59 ± 3 to 38 ± 6 mmHg in FLK) were observed at T1. Arterial hypotension was transient, with normalization of values at T2 and T3. The expired fraction of isoflurane did not differ significantly between the groups. Plasma concentrations of fentanyl, lidocaine and ketamine remained within the therapeutic range. Postoperatively, the number of dogs requiring analgesic rescue was significantly lower in the FLK (0/11, 0%) than in the Fentanyl group (5/11, 45%). In dogs administered morphine and meloxicam as part of the anesthesia protocol, an intraoperative CRI of FLK abolished the requirement for postoperative analgesic rescue for 24 hours in dogs undergoing mastectomy.

In vitro cytotoxic capacity against tumor cell lines and antioxidant activity of acidic polysaccharides isolated from the Andean Patagonian fungus Phylloporia boldo.

Fungal polysaccharides possess a broad biological activity, including cytotoxic and antioxidant activities. This work aimed to evaluate the cytotoxic and antioxidant activity of the acidic polysaccharides of Phylloporia boldo strain (named PBAP40). Cytotoxic activity of polysaccharide was evaluated determining the viability of three tumor cell lines by MTT assay. The effect of acidic polysaccharide on the cell cycle of HL-60 cell line was evaluated by flow cytometry, and the antioxidant activity was determined by DPPH and ABTS assays. PBAP40 showed cytotoxic effects in tumor cell lines. Results suggest that P. boldo acidic polysaccharides arrested tumor cells in the cell cycle Sub G1 phase. The acidic polysaccharides of PBAP40 strain were not cytotoxic for the non-tumor cell line. PBAP40 also showed excellent antioxidant activity. The FT-IR analysis of the acidic polysaccharides indicated the presence of glucans bearing α- and ß- type glycosidic bonds.

Neuroprotective Properties of Eudesmin on a Cellular Model of Amyloid-ß Peptide Toxicity.

BACKGROUND: Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive cognitive impairment and memory loss. One of the hallmarks in AD is amyloid-ß peptide (Aß) accumulation, where the soluble oligomers of Aß (AßOs) are the most toxic species, deteriorating the synaptic function, membrane integrity, and neuronal structures, which ultimately lead to apoptosis. Currently, there are no drugs to arrest AD progression, and current scientific efforts are focused on searching for novel leads to control this disease. Lignans are compounds extracted from conifers and have several medicinal properties. Eudesmin (Eu) is an extractable lignan from the wood of Araucaria araucana, a native tree from Chile. This metabolite has shown a range of biological properties, including the ability to control inflammation and antibacterial effects. OBJECTIVE: In this study, the neuroprotective abilities of Eu on synaptic failure induced by AßOs were analyzed. METHODS: Using neuronal models, PC12 cells, and in silico simulations we evaluated the neuroprotective effect of Eu (30 nM) against the toxicity induced by AßOs. RESULTS: In primary cultures from mouse hippocampus, Eu preserved the synaptic structure against AßOs toxicity, maintaining stable levels of the presynaptic protein SV2 at the same concentration. Eu also averted synapsis failure from the AßOs toxicity by sustaining the frequencies of cytosolic Ca2+ transients. Finally, we found that Eu (30 nM) interacts with the Aß aggregation process inducing a decrease in AßOs toxicity, suggesting an alternative mechanism to explain the neuroprotective activity of Eu. CONCLUSION: We believe that Eu represents a novel lead that reduces the Aß toxicity, opening new research venues for lignans as neuroprotective agents.

Traditional Medicinal Plants as a Source of Inspiration for Osteosarcoma Therapy.

Osteosarcoma is one of the most common types of bone cancers among paediatric patients. Despite the advances made in surgery, chemo-, and radiotherapy, the mortality rate of metastatic osteosarcoma remains unchangeably high. The standard drug combination used to treat this bone cancer has remained the same for the last 20 years, and it produces many dangerous side effects. Through history, from ancient to modern times, nature has been a remarkable source of chemical diversity, used to alleviate human disease. The application of modern scientific technology to the study of natural products has identified many specific molecules with anti-cancer properties. This review describes the latest discovered anti-cancer compounds extracted from traditional medicinal plants, with a focus on osteosarcoma research, and on their cellular and molecular mechanisms of action. The presented compounds have proven to kill osteosarcoma cells by interfering with different pathways: apoptosis induction, stimulation of autophagy, generation of reactive oxygen species, etc. This wide variety of cellular targets confer natural products the potential to be used as chemotherapeutic drugs, and also the ability to act as sensitizers in drug combination treatments. The major hindrance for these molecules is low bioavailability. A problem that may be solved by chemical modification or nano-encapsulation.

Evaluation of Adipose Tissue Zinc-Alpha 2-Glycoprotein Gene Expression and Its Relationship with Metabolic Status and Bariatric Surgery Outcomes in Patients with Class III Obesity.

Zinc-α2 glycoprotein (ZAG) is an adipokine involved in adipocyte metabolism with potential implications in the pathogenesis of metabolic disorders. Our aim was to evaluate the relationship between visceral (VAT) and subcutaneous adipose tissue (SAT) ZAG expression and metabolic parameters in patients with class III obesity, along with the impact of basal ZAG expression on short- and medium-term outcomes related to bariatric surgery. 41 patients with class III obesity who underwent bariatric surgery were included in this study. ZAG gene expression was quantified in SAT and VAT. Patients were classified into two groups according to SAT and VAT ZAG percentile. Anthropometric and biochemical variables were obtained before and 15 days, 45 days, and 1 year after surgery. The lower basal SAT ZAG expression percentile was associated with higher weight and waist circumference, while the lower basal VAT ZAG expression percentile was associated with higher weight, waist circumference, insulin, insulin resistance, and the presence of metabolic syndrome. Basal SAT ZAG expression was inversely related to weight loss at 45 days after surgery, whereas no associations were found between basal VAT ZAG expression and weight loss after surgery. Additionally, a negative association was observed between basal SAT and VAT ZAG expression and the decrease of gamma-glutamyl transferase after bariatric surgery. Therefore, lower SAT and VAT ZAG expression levels were associated with an adverse metabolic profile. However, this fact did not seem to confer worse bariatric surgery-related outcomes. Further research is needed to assess the clinical significance of the role of ZAG expression levels in the dynamics of hepatic enzymes after bariatric surgery.

Sulfated Polysaccharide Extracted from the Green Algae Codium bernabei: Physicochemical Characterization and Antioxidant, Anticoagulant and Antitumor Activity.

Codium bernabei is a green alga that grows on Chilean coasts. The composition of its structural polysaccharides is still unknown. Hence, the aim of this work is to isolate and characterize the hot water extracted polysaccharide fractions. For this purpose, the water extracts were further precipitated in alcohol (TPs) and acid media (APs), respectively. Both fractions were characterized using different physicochemical techniques such as GC-MS, GPC, FTIR, TGA, and SEM. It is confirmed that the extracted fractions are mainly made of sulfated galactan unit, with a degree of sulfation of 19.3% (TPs) and 17.4% (ATs) and a protein content of 3.5% in APs and 15.6% in TPs. Other neutral sugars such as xylose, glucose, galactose, fucose, mannose, and arabinose were found in a molar ratio (0.05:0.6:1.0:0.02:0.14:0.11) for TPs and (0.05:0.31:1.0:0.03:0.1:0.13) for ATs. The molecular weight of the polysaccharide samples was lower than 20 kDa. Both polysaccharides were thermally stable (Tonset > 190 °C) and showed antioxidant activity according to the ABTS•+ and DPPH tests, where TPs fractions had higher scavenging activity (35%) compared to the APs fractions. The PT and APTTS assays were used to measure the anticoagulant activity of the polysaccharide fractions. In general, the PT activity of the TPs and APs was not different from normal plasma values. The exception was the TPs treatment at 1000 µg mL−1 concentration. The APTTS test revealed that clotting time for both polysaccharides was prolonged regarding normal values at 1000 µg mL−1. Finally, the antitumor test in colorectal carcinoma (HTC-116) cell line, breast cancer (MCF-7) and human leukemia (HL-60) cell lines showed the cytotoxic effect of TPs and APs. Those results suggest the potential biotechnological application of sulfate galactan polysaccharides isolated from a Chilean marine resource.

Secretory Profile of Adipose-Tissue-Derived Mesenchymal Stem Cells from Cats with Calicivirus-Positive Severe Chronic Gingivostomatitis.

The feline calicivirus (FCV) causes infections in cats all over the world and seems to be related to a broad variety of clinical presentations, such as feline chronic gingivostomatitis (FCGS), a severe oral pathology in cats. Although its etiopathogeny is largely unknown, FCV infection is likely to be a main predisposing factor for developing this pathology. During recent years, new strategies for treating FCGS have been proposed, based on the use of mesenchymal stem cells (MSC) and their regenerative and immunomodulatory properties. The main mechanism of action of MSC seems to be paracrine, due to the secretion of many biomolecules with different biological functions (secretome). Currently, several pathologies in humans have been shown to be related to functional alterations of the patient's MSCs. However, the possible roles that altered MSCs might have in different diseases, including virus-mediated diseases, remain unknown. We have recently demonstrated that the exosomes produced by the adipose-tissue-derived MSCs (fAd-MSCs) from cats suffering from FCV-positive severe and refractory FCGS showed altered protein contents. Based on these findings, the goal of this work was to analyze the proteomic profile of the secretome produced by feline adipose-tissue-derived MSCs (fAd-MSCs) from FCV-positive patients with FCGS, in order to identify differences between them and to increase our knowledge of the etiopathogenesis of this disease. We used high-resolution mass spectrometry and functional enrichment analysis with Gene Ontology to compare the secretomes produced by the fAd-MSCs of healthy and calicivirus-positive FCGS cats. We found that the fAd-MSCs from cats with FCGS had an increased expression of pro-inflammatory cytokines and an altered proteomic profile compared to the secretome produced by cells from healthy cats. These findings help us gain insight on the roles of MSCs and their possible relation to FCGS, and may be useful for selecting specific biomarkers and for identifying new therapeutic targets.

Suicide gene therapy by canine mesenchymal stem cell transduced with thymidine kinase in a u-87 glioblastoma murine model: Secretory profile and antitumor activity.

The role played by certain domestic species such as dogs as a translational model in comparative oncology shows great interest to develop new therapeutic strategies in brain tumors. Gliomas are a therapeutic challenge that represents the most common form of malignant primary brain tumors in humans and the second most common form in dogs. Gene-directed enzyme/prodrug therapy using adipose mesenchymal stem cells (Ad-MSCs) expressing the herpes simplex virus thymidine kinase (TK) has proven to be a promising alternative in glioblastoma therapy, through its capacity to migrate and home to the tumor and delivering local cytotoxicity avoiding other systemic administration. In this study, we demonstrate the possibility for canine Ad-MSCs (cAd-MSCs) to be genetically engineered efficiently with a lentiviral vector to express TK (TK-cAd-MSCs) and in combination with ganciclovir (GCV) prodrug demonstrated its potential antitumor efficacy in vitro and in vivo in a mice model with the human glioblastoma cell line U87. TK-cAd-MSCs maintained cell proliferation, karyotype stability, and MSCs phenotype. Genetic modification significantly affects its secretory profile, both the analyzed soluble factors and exosomes. TK-cAd-MSCs showed a high secretory profile of some active antitumor immune response cytokines and a threefold increase in the amount of secreted exosomes, with changes in their protein cargo. We also found that the prodrug protein is not released directly into the culture medium by TK-cAd-MSCs. We believe that our work provides new perspectives for glioblastoma gene therapy in dogs and a better understanding of this therapy in view of its possible implantation in humans.

Chitosan-collagen-hydroxyapatite membranes for tissue engineering.

Tissue engineering is growing in developing new technologies focused on providing effective solutions to degenerative pathologies that affect different types of connective tissues. The search for biocompatible, bioactive, biodegradable, and multifunctional materials has grown significantly in recent years. Chitosan, calcium phosphates collagen, and their combination as composite materials fulfill the required properties and could result in biostimulation for tissue regeneration. In the present work, the chitosan/collagen/hydroxyapatite membranes were prepared with different concentrations of collagen and hydroxyapatite. Cell adhesion was evaluated by MTS assay for two in vitro models. Additionally, cytotoxicity of the different membranes employing hemolysis of erythrocytes isolated from human blood was carried out. The structure of the membranes was analyzed by X-rays diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and thermal stability properties by thermogravimetric methods (TGA). The highest cell adhesion after 48 h was obtained for chitosan membranes with the highest hydroxyapatite and collagen content. All composite membranes showed good cell adhesion and low cytotoxicity, suggesting that these materials have a significant potential to be used as biomaterials for tissue engineering. Graphical abstract.

Effect of Canine Adipose Mesenchymal Stem Cell Secretome on a Model of Second-Intention Wound Healing in the Red-Eared Slider Turtle (Trachemys scripta).

Mesenchymal stem cell (MSC) secretome refers to a variety of bioactive compounds that represents the more important pathway by which MSCs participate in tissue regeneration. Many of these compounds have shown variable functional activity even across nonmammalian vertebrate species, although MSCs in turtles have not yet been described. Canine adipose MSC secretome has been successfully used experimentally in skin healing. Our aim was to conduct a blinded controlled study to evaluate the effect of canine adipose MSC secretome (cS-MSC) as an alternative for the healing of soft skin, second intention wounds of red-eared slider turtles (Trachemys scripta). Under general anesthesia, one circular, 6-mm full thickness wound was made in each rear leg of 14 females. After randomization, cS-MSC was injected subcutaneously around one wound at days 1, 7, and 14, whereas the other wound acted as control. Biopsies from three animals' wounds were obtained at days 21, 28, 42, and 63. Differences in mean wound retraction at days 21 (n=14) and 28 (n=11) were statistically nonsignificant. The clinical and histopathologic scores performed blind by two different investigators were similar for treated and control wounds. In conclusion, we could not detect a significant functional activity of cS-MSC on wound healing of Trachemys scripta.

Oridonin enhances antitumor effects of doxorubicin in human osteosarcoma cells.

BACKGROUND: Doxorubicin is the chemotherapeutic drug of choice in osteosarcoma treatment, but its cumulative administration causes dilated cardiomyopathy. Combination therapy represents a potential strategy to reduce the therapeutic dosage of the chemotherapeutic agent and minimize its side effects. The aim of this study was to evaluate the potential of oridonin, a natural product from the medicinal herb Rabdosia rubescens, to act in combination with doxorubicin for osteosarcoma treatment. To date, there are no reports of the simultaneous administration of both drugs in osteosarcoma therapy. METHODS: The combined administration of different doses of oridonin and doxorubicin, as compared with the drugs alone, were tested in an in vitro model of osteosarcoma. The synergistic effect of the drugs on cell death was assessed by alamarBlue™ and by CompuSyn software. Early and late apoptosis markers (JC-1 fluorescence and Annexin V immunofluorescence), as well as the production of reactive oxygen species, were evaluated by flow cytometry. Western blot was used to assess the expression of anti-apoptotic proteins. RESULTS: Oridonin and doxorubicin presented a synergistic cytotoxic effect in osteosarcoma cells. In the presence of sub-cytotoxic concentrations of the natural product, there was an increased accumulation of intracellular doxorubicin, increased levels of reactive oxygen species (ROS), alteration of mitochondria membrane potential and a higher rate of apoptosis. CONCLUSION: The combined use of oridonin and doxorubicin could help to reduce the clinical dosage of doxorubicin and its dangerous side effects.

High antioxidant activity of phenolic compounds dampens oxidative stress in Espinosa nothofagi galls induced on Nothofagus obliqua buds.

Reactive oxygen species (ROS) are considered the first signaling molecules involved in gall development, linked to the establishment of cyto-histological gradients leading to gall tissue redifferentiation. ROS overproduction induces the failure of gall establishment or its premature senescence. Galls could therefore have efficient mechanisms of ROS dissipation and maintenance of homeostasis, such as polyphenol synthesis. The co-occurrence of ROS and polyphenols in the Espinosa nothofagi galls induced on Nothofagus obliqua buds was explored and was related to the antioxidant capacity of the inner (IC) and outer (OC) gall compartments. We hypothesize that: (i) ROS are produced and accumulated in both tissue compartments of E. nothofagi galls in co-occurrence with polyphenolic, flavonols, and lignin, conferring high antioxidant activity to inner and outer gall tissue compartment; (ii) antioxidant activity is higher in IC related to a higher polyphenol concentration in this compartment. The results show that ROS and polyphenols, mainly flavonols, are produced and accumulated in IC and OC, while lignin accumulated mainly in the IC. In both gall compartments, polyphenols mediate ROS elimination, confirmed by histochemical and spectrophotometry techniques. The IC extract has the highest antioxidant capacity, probably due to lignin deposition and a higher polyphenol concentration in this compartment.

Epidural administration of combinations of ropivacaine, morphine and xylazine in bitches undergoing total unilateral mastectomy: a randomized clinical trial.

OBJECTIVE: To investigate the epidural administration of combinations of ropivacaine, morphine and xylazine in bitches undergoing unilateral mastectomy. STUDY DESIGN: Prospective, randomized, blinded, clinical study. ANIMALS: A total of 22 bitches scheduled to undergo unilateral mastectomy for mammary tumor excision. METHODS: Dogs were anesthetized with acepromazine (0.02 mg kg-1) and morphine (0.3 mg kg-1) intramuscularly, propofol intravenously (IV) and isoflurane. Prior to the beginning of surgery, dogs were randomly administered one of three epidural treatments: ropivacaine (0.75 mg kg-1) with morphine (0.1 mg kg-1) (group RM, n = 7); ropivacaine with xylazine (0.1 mg kg-1) (group RX, n = 8); or ropivacaine with morphine and xylazine (group RMX, n = 7). Cardiopulmonary variables and the expired concentration of isoflurane (Fe'Iso) were recorded intraoperatively. Meloxicam (0.1 mg kg-1) was administered IV during skin closure. Postoperative pain scores were evaluated with the Glasgow composite measure pain scale short form for 24 hours, and rescue analgesia with morphine (0.5 mg kg-1) was administered intramuscularly when pain scores were ≥ 6/24. RESULTS: Fe'Iso was significantly higher in group RM than in groups RX and RMX. Heart rate decreased significantly in groups RX and RMX, but blood pressure remained within acceptable values. The number of dogs administered rescue analgesia within 24 hours was significantly higher in group RX (seven dogs, 87.5%) than in groups RM (one dog, 14.3%; p = 0.01) and RMX (two dogs, 28.6%; p = 0.04). Time to standing was significantly longer in group RX than in group RM. CONCLUSIONS AND CLINICAL RELEVANCE: All epidural treatments provided adequate antinociception with minimal cardiovascular adverse effects during mastectomy. The inclusion of morphine (groups RM and RMX) provided the best postoperative analgesia. Owing to the undesirable effect of xylazine on ambulation, the combination ropivacaine-morphine appeared to provide greater benefits in bitches undergoing unilateral mastectomy.

Inflammation, a common mechanism in frailty and COVID-19, and stem cells as a therapeutic approach.

As our life expectancy increases, specific medical conditions appear, and new challenges are met in terms of global health. Frailty has become a medical and scientific concept to define pathologies where inflammation, depressed immune system, cellular senescence, and molecular aging converge. But more importantly, frailty is the ultimate cause of death that limits our life span and deteriorates health in an increasing proportion of the world population. The difficulty of tackling this problem is the combination of factors that influence frailty appearance, such as stem cells exhaustion, inflammation, loss of regeneration capability, and impaired immunomodulation. To date, multiple research fields have found mechanisms participating in this health condition, but to make progress, science will need to investigate frailty with an interdisciplinary approach. This article summarizes the current efforts to understand frailty from their processes mediated by inflammation, aging, and stem cells to provide a new perspective that unifies the efforts in producing advanced therapies against medical conditions in the context of frailty. We believe this approach against frailty is particularly relevant to COVID-19, since people in a state of frailty die more frequently due to the hyperinflammatory process associated with this infection.

Renal scintigraphy as an early and efficient method for detecting loss of renal function in a cat.

CASE SUMMARY: A 6-year-old mixed-breed male cat was evaluated for a routine annual health assessment. No alterations on physical examination were observed other than mild pain on palpation of the right kidney. Complete blood count, serum biochemistry (including symmetric dimethylarginine), urinalysis and urine protein:creatinine ratio were within the reference intervals for the species. Abdominal ultrasonography showed the presence of asymmetric kidneys, decreased corticomedullary definition, presence of a cyst on the left kidney and moderate renal pelvis dilatation on the right kidney. Dynamic renal scintigraphy (technetium [99mTc]-diethylenetriamine pentaacetic acid) revealed a single functioning kidney on the left. Static renal scintigraphy (99mTc-dimercaptosuccinic acid) exhibited renal activity practically restricted to the left kidney (relative uptake was 99% for the left kidney and 1% for the right kidney). Results of renal scintigraphy showed that the left kidney was compensating for the lack of function of the right one. GFR was 2.17 ml/min/kg, which is considered subclinical renal insufficiency and is in accordance with the case, as the cat was asymptomatic and did not present alterations in laboratory parameters. RELEVANCE AND NOVEL INFORMATION: Renal scintigraphy was an important tool to determine the loss of renal function in one of the kidneys and mild reduction of global GFR. In this case report, renal scintigraphy proved to be more sensitive in the assessment of renal function than other tests routinely performed.

Antiproliferative activity of carotenoid pigments produced by extremophile bacteria.

Various microorganisms are able to synthesize pigments, which usually present antioxidant properties. The aim of this work was to evaluate the antiproliferative activity of bacterial pigments against cancer cells Neuro-2a, Saos-2 and MCF-7. Pigments were obtained from Deinococcus sp. UDEC-P1 and Arthrobacter sp. UDEC-A13. Both bacterial strains were isolated from cold environments (Patagonia and Antarctica, respectively). Pigments were purified and analyzed by HPLC. Antiproliferative activity was evaluated by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium (MTT) assay. Deinoxanthin carotenoid obtained from Deinococcus sp. UDEC-P1 was able to reduce significatively the viability of Saos-2 (37.1%), while no effect was observed against MCF-7 and Neuro-2a. Pigments obtained from Arthrobacter sp. UDEC-A13 showed a significant viability reduction of three tumour cells (20.6% Neuro-2a, 26.3% Saos-2 and 13.2% MCF-7). Therefore, carotenoid pigments produced by extremophilic bacteria Deinococcus sp. UDEC-P1 and Arthrobacter sp. UDEC-A13 could be proposed as novel complementary compounds in anticancer chemotherapy.