Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis.

High-resolution respirometry (HRR) can assess peripheral blood mononuclear cell (PBMC) bioenergetics, but no standardized medium for PBMC preparation and HRR analysis exist. Here, we study the effect of four different media (MiR05, PBS, RPMI, Plasmax) on the count, size, and HRR (Oxygraph-O2k) of intact PBMCs. Remarkably, the cell count was 21 % higher when PBMCs were resuspended in MiR05 than in PBS or Plasmax, causing O2 flux underestimation during HRR due to inherent adjustments. Moreover, smaller cell size and cell aggregation was observed in MiR05. Based on our findings, we propose that Plasmax, PBS or RPMI is more suitable than MiR05 for HRR of intact PBMCs. We provide oxygen solubility factors for Plasmax and PBS and encourage further optimization of a standardized HRR protocol for intact PBMCs.

Effect of Parenteral Supplementation of Minerals and Vitamins on Oxidative Stress Biomarkers and Hepatic Fatty Acid Metabolism in Dairy Cows During the Transition Period.

In the present work we aimed to study the effects of parenteral vitamin and mineral supplementation on hepatic fatty acid metabolism as well as on the oxidative stress biomarkers in biological samples of transition cows. The supplemented group (SG, n = 11) received a subcutaneous injection of 5 mL of vitamin A palmitate 35 mg/mL, vitamin E acetate 50 mg/mL plus other injection of 5 mL of copper edetate 10 mg/mL, zinc edetate 40 mg/mL, manganese edetate 10 mg/mL, and sodium selenite 5 mg/mL on days - 60, - 30, and 7 (± 3) relative to calving. The control group (CG, n = 11) received two subcutaneous injections of 5 mL of 9 mg/mL sodium chloride at the same times of the SG. Blood, urine, and liver biopsies were sampled 21 (± 3) days before the expected calving date and 7 and 21 (± 3) days after calving. Results revealed that supplemented animals had higher glutation peroxidase (GSH-Px) activity, lower and higher concentration of 3-nitrotyrosine (3-NT) in the liver and plasma, respectively, higher expression of the mitochondrial beta-oxidation enzyme carnitine palmitoyltransferase 1 in the liver, and lower content of hepatic triacylglycerol, mirroring plasma liver function parameters. No differences between groups were found in the superoxide dismutase activity, MDA concentrations, the protein abundance of peroxisomal acyl-CoA oxidase 1, diacylglycerol O-acyltransferase 1, and peroxisome proliferator-activated receptor alpha. These results suggest that the vitamin and mineral supplementation provided to dairy cows had a beneficial effect on GSH-Px activity, hepatic 3-NT concentration, and on the metabolic adaptation during the peripartum period.

Cytotoxic, antioxidant, and cytoprotective properties of polyphenol-enriched extracts from pecan nutshells in MDA-MB-231 breast cancer cells.

Phenolic compounds present in plants have demonstrated several biological properties such as antioxidant, antitumor, cardioprotective, and antiproliferative. On the other hand, doxorubicin, a chemotherapeutic widely used to treat breast cancer, usually exhibits chronic cardiotoxicity associated with oxidative stress. Therefore, we aimed to study the effects of phenolic compound-enriched extract (PCEE) with doxorubicin in breast cancer. To achieve this, after an SPE-C18 -column purification process of crude extracts obtained from pecan nutshells (Carya illinoinensis), the resulting PCEE was used to evaluate the cytotoxicity and antioxidant properties against the human breast cancer cell line MDA-MB-231 and the normal-hamster ovary cell line CHO-K1. PCEE was selectively cytotoxic against both cell lines, with an IC50 value (≈26.34 mg/L) for MDA-MB-231 lower than that obtained for CHO-K1 (≈55.63 mg/L). As a cytotoxic mechanism, PCEE inhibited cell growth by G2/M cell cycle arrest in MDA-MB-231 cells. Simultaneously, the study of the antioxidant activity showed that PCEE had a cytoprotective effect, evidenced by reduced ROS production in cells with oxidative stress caused by doxorubicin. The results highlight PCEE as a potential antitumor agent, thus revaluing it as an agro-industrial residue.

Peripheral blood mononuclear cells exhibit increased mitochondrial respiration after adjuvant chemo- and radiotherapy for early breast cancer.

BACKGROUND: Adjuvant chemo- and radiotherapy cause cellular damage to tumorous and healthy dividing cells. Chemotherapy has been shown to cause mitochondrial respiratory dysfunction in non-tumorous tissues, but the effects on human peripheral blood mononuclear cells (PBMCs) remain unknown. AIM: We aimed to investigate mitochondrial respiration of PBMCs before and after adjuvant chemo- and radiotherapy in postmenopausal patients with early breast cancer (EBC) and relate these to metabolic parameters of the patients. METHODS: Twenty-three postmenopausal women diagnosed with EBC were examined before and shortly after chemotherapy with (n = 18) or without (n = 5) radiotherapy. Respiration (O2 flux per million PBMCs) was assessed by high-resolution respirometry of intact and permeabilized PBMCs. Clinical metabolic characteristics and mitochondrial DNA (mtDNA) content of PBMCs (mtDN relative to nuclear DNA) were furthermore assessed. RESULTS: Respiration of intact and permeabilized PBMCs from EBC patients significantly increased with adjuvant chemo- and radiotherapy (p = 6 × 10-5 and p = 1 × 10-7 , respectively). The oxygen flux attributed to specific mitochondrial complexes and respiratory states increased by 17-43% compared to before therapy initiation. Similarly, PBMC mtDNA content increased by 40% (p = 0.002). Leukocytes (p = 0.0001), hemoglobin (p = 0.0003), and HDL cholesterol (p = 0.003) concentrations decreased whereas triglyceride (p = 0.01) and LDL (p = 0.02) concentrations increased after treatment suggesting a worsened metabolic state. None of the metabolic parameters or the mtDNA content of PBMCs correlated significantly with PBMC respiration. CONCLUSION: This study shows that mitochondrial respiration and mtDNA content in circulating PBMCs increase after adjuvant chemo- and radiotherapy in postmenopausal patients with EBC. Besides the increased mtDNA content, a shift in PBMC subpopulation proportions towards cells relying on oxidative phosphorylation, who may be less sensitive to chemotherapy, might influence the increased mitochondrial respiration observed iafter chemotherapy.

Biomarkers of oxidative stress and liver function in early lactation and their relationship with the reproductive efficiency of multiparous grazing dairy cows in Argentina. A retrospective study.

This study aimed to analyze the possible relationship between days to conception and different oxidative stress (OS) biomarkers and liver functional parameters in multiparous dairy cows. Besides, a fast reliable method for the accurate measurement of malondialdehyde (MDA) by liquid chromatography-tandem mass spectrometry was developed in several matrices. During lactation, the days to conception of 28 cows were determined for a retrospective study. According to this parameter, cows were divided into two groups: high and low days to conception (HDC and LDC, respectively). Blood, urine and liver biopsies were sampled 21 days before the expected calving date, and 7 and 21 days after calving. The method developed for MDA was validated according to international requirements. The lower limit of quantification was 0.25 µmol/L for plasma and urine and 10.00 µmol/L for liver tissue. No differences between groups were observed in the systemic concentration of non-esterified fatty acids, ß-hydroxybutyric acid and liver triacylglycerol content (P > 0.05). Cholesterol concentration was higher in the LDC than in the HDC group (P < 0.05). Plasma 3-nitrotyrosine (3-NT) concentration was lower in the LDC than in the HDC group on day 21 post-calving (P < 0.05). Superoxide dismutase activity was higher in the LDC than in the HDC group (P < 0.05). Particularly, in the liver, 3-NT and MDA concentrations were lower in the LDC than in the HDC group (P < 0.05). These results allow inferring that the amelioration of OS biomarkers in plasma and liver could be related to a better reproductive performance of dairy cows.

Extraction of phenolic compounds from the shells of pecan nuts with cytotoxic activity through apoptosis against the colon cancer cell line HT-29.

The water extraction of phenolic compounds from two varieties ("Mahan" and "Marameck") of pecan nutshells (Carya illinoinensis) without and with sonication, varying the solvent/solid ratio (S), the pH, and the refluxing time (t), was studied. Additionally, the in vitro cytotoxicity and the determination of the cell death mechanism of the extracts against the colon cancer cell line HT-29 were investigated. The content of total phenolic compounds (TPC) of "Marameck" nutshells resulted higher than for the "Mahan" variety, and the pH increase resulted in higher TPC contents for both cultivars. The optimized conditions for TPC extraction without and with sonication resulted: S = 33 ml/g, pH = 12, and t = 9.6 min, and yielded ≈ 70 and 90 mg/g of TPC for "Mahan" and "Marameck" nutshells, respectively. The optimized extracts of pecan nutshells without sonication from both cultivars presented similar cytotoxicity against HT-29 colon cancer cells (IC50  ≈ 50 µg/ml), higher than for sonicated extracts (IC50 ≈ 88 and 138 µg/ml for "Mahan" and "Marameck," respectively). Cell death through apoptosis was the main mechanism of cell death induced by the nutshell extracts. PRACTICAL APPLICATION: The extraction of phenolic compounds (TPC) from the residues of two varieties of pecan nutshells ("Mahan" and "Marameck") was studied. An optimal combination of variables within the pH range that minimizes the solvent-to-solid ratio (S) and the time of refluxing (t), saving at the same time, water and energy, was set up. The phenolic compound extracts obtained from the residues of the pecan nuts exhibit cytotoxic effects against colon cancer cells and could be of interest as an alternative treatment of different types of cancer. Additionally, these extracts may be of importance to the food industry as they can be used as antioxidant agents in food formulation. Also, the high levels of anthocyanidins obtained from the pecan nut extracts after proanthocyanidins' strong acid hydrolysis can be purified and employed as natural red dyes.

Social interaction layers in complex networks for the dynamical epidemic modeling of COVID-19 in Brazil.

We are currently living in a state of uncertainty due to the pandemic caused by the SARS-CoV-2 virus. There are several factors involved in the epidemic spreading, such as the individual characteristics of each city/country. The true shape of the epidemic dynamics is a large, complex system, considerably hard to predict. In this context, Complex networks are a great candidate for analyzing these systems due to their ability to tackle structural and dynamic properties. Therefore, this study presents a new approach to model the COVID-19 epidemic using a multi-layer complex network, where nodes represent people, edges are social contacts, and layers represent different social activities. The model improves the traditional SIR, and it is applied to study the Brazilian epidemic considering data up to 05/26/2020, and analyzing possible future actions and their consequences. The network is characterized using statistics of infection, death, and hospitalization time. To simulate isolation, social distancing, or precautionary measures, we remove layers and reduce social contact's intensity. Results show that even taking various optimistic assumptions, the current isolation levels in Brazil still may lead to a critical scenario for the healthcare system and a considerable death toll (average of 149,000). If all activities return to normal, the epidemic growth may suffer a steep increase, and the demand for ICU beds may surpass three times the country's capacity. This situation would surely lead to a catastrophic scenario, as our estimation reaches an average of 212,000 deaths, even considering that all cases are effectively treated. The increase of isolation (up to a lockdown) shows to be the best option to keep the situation under the healthcare system capacity, aside from ensuring a faster decrease of new case occurrences (months of difference), and a significantly smaller death toll (average of 87,000).

Electrochemical and optical detection and machine learning applied to images of genosensors for diagnosis of prostate cancer with the biomarker PCA3.

The development of simple detection methods aimed at widespread screening and testing is crucial for many infections and diseases, including prostate cancer where early diagnosis increases the chances of cure considerably. In this paper, we report on genosensors with different detection principles for a prostate cancer specific DNA sequence (PCA3). The genosensors were made with carbon printed electrodes or quartz coated with layer-by-layer (LbL) films containing gold nanoparticles and chondroitin sulfate and a layer of a complementary DNA sequence (PCA3 probe). The highest sensitivity was reached with electrochemical impedance spectroscopy with the detection limit of 83 pM in solutions of PCA3, while the limits of detection were 2000 pM and 900 pM for cyclic voltammetry and UV-vis spectroscopy, respectively. That detection could be performed with an optical method is encouraging, as one may envisage extending it to colorimetric tests. Since the morphology of sensing units is known to be affected in detection experiments, we applied machine learning algorithms to classify scanning electron microscopy images of the genosensors and managed to distinguish those exposed to PCA3-containing solutions from control measurements with an accuracy of 99.9%. The performance in distinguishing each individual PCA3 concentration in a multiclass task was lower, with an accuracy of 88.3%, which means that further developments in image analysis are required for this innovative approach.

Accessory muscle of the flexor digitorum superficialis and its clinical implications.

Anatomical variations of the flexor digitorum superficialis (FDS) muscle and tendon unit are frequently reported by anatomists and clinicians. Anatomical muscle variations of the FDS and its tendons may include variations of muscle belly, presence of accessory or duplicate tendons, abnormal tendon connections, and absence of muscle or tendon components. Such variations may or may not have clinical implications. This report presents a case not described previously: a unilateral accessory muscle of the flexor digitorum superficialis which was connected by a thick tendon to the flexor digitorum superficialis muscle; it was directed proximally to the insertion of the medial epicondyle of the humerus, next to the superficialis head of the pronator teres muscle. The belly of the accessory muscle was positioned anterior to the median and anterior interosseous nerve. This anatomical variation is known as type V in the classification of Elliot et al. The knowledge of these anatomical variations helps hand surgeons interpret the clinical examination, particularly in the evaluation of patients who have suffered tendon injuries or show sign s of possible peripheral nerve entrapment.

Variações anatômicas (anomalias) da unidade musculotendínea do flexor superficial dos dedos (FSD) têm sido relatadas com frequência na literatura em tratados, artigos clínicos e anatômicos. Podem ocorrer variações do corpo muscular, presença de tendões acessórios ou duplicados, conexões musculotendinosas anormais e ausência do componente muscular ou tendinoso. Essas variações podem ou não ter implicações clínicas. Os autores apresentam um caso não descrito previamente de um músculo acessório do músculo FSD unilateral que estava conectado através de um tendão espesso ao músculo FSD e dirigia-se proximalmente para inserir-se no epicôndilo medial do úmero ao lado da cabeça superficial do músculo pronador redondo. O músculo flexor superficial acessório posicionava anteriormente aos nervos mediano e interósseo anterior. Essa variação se enquadra no tipo V da classificação de Elliot et al. O conhecimento dessas variações anatômicas auxilia o cirurgião da mão a interpretar o exame clínico, em especial na avaliação de pacientes que sofreram lesões tendinosas ou apresentam sinais de possíveis compressão de algum nervo periférico.

Accessory muscle of the flexor digitorum superficialis and its clinical implications / Músculo acessório do músculo flexor superficial e sua implicação clínica

ABSTRACT Anatomical variations of the flexor digitorum superficialis (FDS) muscle and tendon unit are frequently reported by anatomists and clinicians. Anatomical muscle variations of the FDS and its tendons may include variations of muscle belly, presence of accessory or duplicate tendons, abnormal tendon connections, and absence of muscle or tendon components. Such variations may or may not have clinical implications. This report presents a case not described previously: a unilateral accessory muscle of the flexor digitorum superficialis which was connected by a thick tendon to the flexor digitorum superficialis muscle; it was directed proximally to the insertion of the medial epicondyle of the humerus, next to the superficialis head of the pronator teres muscle. The belly of the accessory muscle was positioned anterior to the median and anterior interosseous nerve. This anatomical variation is known as type V in the classification of Elliot et al. The knowledge of these anatomical variations helps hand surgeons interpret the clinical examination, particularly in the evaluation of patients who have suffered tendon injuries or show sign s of possible peripheral nerve entrapment.

RESUMO Variações anatômicas (anomalias) da unidade musculotendínea do flexor superficial dos dedos (FSD) têm sido relatadas com frequência na literatura em tratados, artigos clínicos e anatômicos. Podem ocorrer variações do corpo muscular, presença de tendões acessórios ou duplicados, conexões musculotendinosas anormais e ausência do componente muscular ou tendinoso. Essas variações podem ou não ter implicações clínicas. Os autores apresentam um caso não descrito previamente de um músculo acessório do músculo FSD unilateral que estava conectado através de um tendão espesso ao músculo FSD e dirigia-se proximalmente para inserir-se no epicôndilo medial do úmero ao lado da cabeça superficial do músculo pronador redondo. O músculo flexor superficial acessório posicionava anteriormente aos nervos mediano e interósseo anterior. Essa variação se enquadra no tipo V da classificação de Elliot et al. O conhecimento dessas variações anatômicas auxilia o cirurgião da mão a interpretar o exame clínico, em especial na avaliação de pacientes que sofreram lesões tendinosas ou apresentam sinais de possíveis compressão de algum nervo periférico.