Liposomal quercetin: A promising strategy to combat hepatic insulin resistance and inflammation in type 2 diabetes mellitus.

In type 2 diabetes mellitus, hepatic insulin resistance is intricately associated with oxidative stress and inflammation. Nonetheless, the lack of therapeutic interventions directly targeting hepatic dysfunction represents a notable gap in current treatment options. Flavonoids have been explored due to their potential antidiabetic effects. However, these compounds are associated with low bioavailability and high metabolization. In the present study, four flavonoids, kaempferol, quercetin, kaempferol-7-O-glucoside and quercetin-7-O-glucoside, were studied in a cellular model of hepatic insulin resistance using HepG2 cells. Quercetin was selected as the most promising flavonoid and incorporated into liposomes to enhance its therapeutic effect. Quercetin liposomes had a mean size of 0.12 µm, with an incorporation efficiency of 93 %. Quercetin liposomes exhibited increased efficacy in modulating insulin resistance. This was achieved through the modulation of Akt expression and the attenuation of inflammation, particularly via the NF-κB pathway, as well as the regulation of PGE2 and COX-2 expression. Furthermore, quercetin liposomes displayed a significant advantage over free quercetin in attenuating the production of reactive pro-oxidant species. These findings open new avenues for developing innovative therapeutic strategies to manage diabetes, emphasizing the potential of quercetin liposomes as a promising approach for targeting both hepatic insulin resistance and associated inflammation.

Autophagy is the main driver of radioresistance of HNSCC cells in mild hypoxia.

Hypoxia poses a significant challenge to the effectiveness of radiotherapy in head and neck squamous cell carcinoma (HNSCC) patients, and it is imperative to discover novel approaches to overcome this. In this study, we investigated the underlying mechanisms contributing to x-ray radioresistance in HPV-negative HNSCC cells under mild hypoxic conditions (1% oxygen) and explored the potential for autophagy modulation as a promising therapeutic strategy. Our findings show that HNSCC cells exposed to mild hypoxic conditions exhibit increased radioresistance, which is largely mediated by the hypoxia-inducible factor (HIF) pathway. We demonstrate that siRNA knockdown of HIF-1α and HIF-1ß leads to increased radiosensitivity in HNSCC cells under hypoxia. Hypoxia-induced radioresistance was not attributed to differences in DNA double strand break repair kinetics, as these remain largely unchanged under normoxic and hypoxic conditions. Rather, we identify autophagy as a critical protective mechanism in HNSCC cells following irradiation under mild hypoxia conditions. Targeting key autophagy genes, such as BECLIN1 and BNIP3/3L, using siRNA sensitizes these cells to irradiation. Whilst autophagy's role in hypoxic radioresistance remains controversial, this study highlights the importance of autophagy modulation as a potential therapeutic approach to enhance the effectiveness of radiotherapy in HNSCC.

Synopsis of the species of Ortholinea Shulman, 1962 (Cnidaria: Myxosporea: Ortholineidae).

A synopsis of Ortholinea Shulman, 1962 (Cnidaria: Myxosporea: Ortholineidae) is presented and identifies 26 nominal species presently allocated within this genus. Species morphological and morphometric features, tissue tropism, type-host, and type-locality are provided from original descriptions. Data from subsequent redescriptions and reports is also given. Accession numbers to sequences deposited in GenBank are indicated when available, and the myxospores were redrawn based on original descriptions. The information gathered shows that Ortholinea infect a wide taxonomic variety of freshwater and marine fish. Nonetheless, the broad host specificity reported for several species is not fully supported by morphological descriptions and requires molecular corroboration. The members of this genus are coelozoic and mainly parasitize the urinary system, with few species occurring in the gallbladder. Ortholinea visakhapatnamensis is the only exception, being histozoic in the visceral peritoneum. Molecular data of the small subunit ribosomal RNA gene (SSU rDNA) is available for about one third of Ortholinea species, with genetic interspecific variation ranging between 1.65% and 29.1%. Phylogenetic analyses reveal Ortholinea to be polyphyletic, with available SSU rDNA sequences clustering within the subclades of the highly heterogenous freshwater urinary clade of the oligochaete-infecting lineage. The life cycles of two Ortholinea species have been clarified based on molecular inferences and identify triactinomyxon actinospores as counterparts, and marine oligochaetes of the family Naididae as permissive hosts to this genus.

Global acetylome profiling indicates EPA impedes but OA promotes prostate cancer motility through altered acetylation of PFN1 and FLNA.

Prostate cancer (PCa) is one of the leading causes of cancer morbidity and mortality in men. Metastasis is the main cause of PCa-associated death. Recent evidence indicated a significant reduction in PCa mortality associated with higher ω-3 polyunsaturated fatty acids (PUFAs) consumption. However, the underlying mechanisms remained elusive. In this study, we applied global acetylome profiling to study the effect of fatty acids treatment. Results indicated that oleic acid (OA, monounsaturated fatty acid, MUFA, 100 µM) elevates while EPA (eicosapentaenoic acid, 100 µM) reduces the acetyl-CoA level, which alters the global acetylome. After treatment, two crucial cell motility regulators, PFN1 and FLNA, were found with altered acetylation levels. OA increased the acetylation of PFN1 and FLNA, whereas EPA decreased PFN1 acetylation level. Furthermore, OA promotes while EPA inhibits PCa migration and invasion. Immunofluorescence assay indicated that EPA impedes the formation of lamellipodia or filopodia through reduced localization of PFN1 and FLNA to the leading edge of cells. Therefore, perturbed acetylome may be one critical step in fatty acid-affected cancer cell motility. This study provides some new insights into the response of ω-3 PUFAs treatment and a better understanding of cancer cell migration and invasion modulation.

Enhancing radiotherapy outcomes in rectal cancer: A systematic review of targeting hypoxia-induced radioresistance.

Introduction: Neoadjuvant radiotherapy is successfully used in rectal cancer to improve overall survival. However, treatment response is both unpredictable and variable. There is strong evidence to show that the phenomenon of tumour hypoxia is associated with radioresistance, however the mechanism(s) behind this are poorly understood. Consequently, there have only been a small number of studies evaluating methods targeting hypoxia-induced radioresistance. The purpose of this systematic review is to evaluate the potential effectiveness of targeting hypoxia-induced radioresistance in rectal cancer and provide recommendations for future research in this area. Methods: A comprehensive literature search was performed following the PRISMA guidelines. This study was registered on the Prospero database (CRD42023441983). Results: Eight articles met the inclusion criteria. All studies identified were in vitro or in vivo studies, there were no clinical trials. Of the 8 studies identified, 5 assessed the efficacy of drugs which directly or indirectly targeted hypoxia and three that identified potential targets. There was conflicting in vivo evidence for the use of metformin to overcome hypoxia induced radioresistance. Vorinostat, atovaquone, and evofosfamide showed promising preclinical evidence that they can overcome hypoxia-induced radioresistance. Discussion: The importance of investigating hypoxia-induced radioresistance in rectal cancer is crucial. However, to date, only a small number of preclinical studies exist evaluating this phenomenon. This systematic review highlights the importance of further research to fully understand the mechanism behind this radioresistance. There are promising targets identified in this systematic review however, substantially more pre-clinical and clinical research as a priority for future research is needed.

The role of autophagy in hypoxia-induced radioresistance.

Radiotherapy is a widely used treatment modality against cancer, and although survival rates are increasing, radioresistant properties of tumours remain a significant barrier for curative treatment. Tumour hypoxia is one of the main contributors to radioresistance and is common in most solid tumours. Hypoxia is responsible for many molecular changes within the cell which helps tumours to survive under such challenging conditions. These hypoxia-induced molecular changes are predominantly coordinated by the hypoxia inducible factor (HIF) and have been linked with the ability to confer resistance to radiation-induced cell death. To overcome this obstacle research has been directed towards autophagy, a cellular process involved in self degradation and recycling of macromolecules, as HIF plays a large role in its coordination under hypoxic conditions. The role that autophagy has following radiotherapy treatment is conflicted with evidence of both cytoprotective and cytotoxic effects. This literature review aims to explore the intricate relationship between radiotherapy, hypoxia, and autophagy in the context of cancer treatment. It provides valuable insights into the potential of targeting autophagy as a therapeutic strategy to improve the response of hypoxic tumours to radiotherapy.

Oxygen-regulated post-translation modifications as master signalling pathway in cells.

Oxygen is essential for viability in mammalian organisms. However, cells are often exposed to changes in oxygen availability, due to either increased demand or reduced oxygen supply, herein called hypoxia. To be able to survive and/or adapt to hypoxia, cells activate a variety of signalling cascades resulting in changes to chromatin, gene expression, metabolism and viability. Cellular signalling is often mediated via post-translational modifications (PTMs), and this is no different in response to hypoxia. Many enzymes require oxygen for their activity and oxygen can directly influence several PTMS. Here, we review the direct impact of changes in oxygen availability on PTMs such as proline, asparagine, histidine and lysine hydroxylation, lysine and arginine methylation and cysteine dioxygenation, with a focus on mammalian systems. In addition, indirect hypoxia-dependent effects on phosphorylation, ubiquitination and sumoylation will also be discussed. Direct and indirect oxygen-regulated changes to PTMs are coordinated to achieve the cell's ultimate response to hypoxia. However, specific oxygen sensitivity and the functional relevance of some of the identified PTMs still require significant research.

Comprehensive analysis of the lysine succinylome in fish oil-treated prostate cancer cells.

Prostate cancer (PCa) poses a significant health threat to males, and research has shown that fish oil (FO) can impede PCa progression by activating multiple mitochondria-related pathways. Our research is focused on investigating the impact of FO on succinylation, a posttranslational modification that is closely associated with mitochondria in PCa cells. This study employed a mass spectrometry-based approach to investigate succinylation in PCa cells. Bioinformatics analysis of these succinylated proteins identified glutamic-oxaloacetic transaminase 2 (GOT2) protein as a key player in PCa cell proliferation. Immunoprecipitation and RNA interference technologies validated the functional data. Further analyses revealed the significance of GOT2 protein in regulating nucleotide synthesis by providing aspartate, which is critical for the survival and proliferation of PCa cells. Our findings suggest that FO-dependent GOT2 succinylation status has the potential to inhibit building block generation. This study lays a solid foundation for future research into the role of succinylation in various biological processes. This study highlights the potential use of FO as a nutrition supplement for managing and slowing down PCa progression.

Targeting SOD1 via RNAi with PEGylated graphene oxide nanoparticles in platinum-resistant ovarian cancer.

Acquired platinum resistance poses a significant therapeutic impediment to ovarian cancer patient care, accounting for more than 200,000 deaths annually worldwide. We previously identified that overexpression of the antioxidant superoxide dismutase 1 (SOD1) in ovarian cancer is associated with a platinum-resistant phenotype via conferring oxidative stress resistance against platinum compounds. We further demonstrated that enzymatic inhibition using small-molecule inhibitors or silencing of SOD1 via RNA interference (RNAi) increased cisplatin sensitivity and potency in vitro. We launched this study to explore the potential therapeutic applications of SOD1 silencing in vivo in order to reverse cisplatin resistance using a graphene-based siRNA delivery platform. PEGylated graphene oxide (GO) polyethyleneimine (GOPEI-mPEG) nanoparticle was complexed with SOD1 siRNA. GOPEI-mPEG-siSOD1 exhibited high biocompatibility, siRNA loading capacity, and serum stability, and showed potent downregulation of SOD1 mRNA and protein levels. We further observed that cisplatin and PEI elicited mitochondrial dysfunction and transcriptionally activated the mitochondrial unfolded protein response (UPRmt) used as a reporter for their respective cytotoxicities. SOD1 silencing was found to augment cisplatin-induced cytotoxicity resulting in considerable tumour growth inhibition in cisplatin-sensitive A2780 and cisplatin-resistant A2780DDP subcutaneous mouse xenografts. Our study highlights the potential therapeutic applicability of RNAi-mediated targeting of SOD1 as a chemosensitizer for platinum-resistant ovarian cancers.

Frequency of Autoantibodies on Non-Hodgkin Lymphoma.

(1) Background: Non-Hodgkin Lymphoma is a neoplasm that can significantly compromise the immune system, but timely assessment can change the patient outcome. In cancer, the activation of the immune system could lead to the secretion of autoantibodies. (2) Methods: A retrospective cohort study was performed from 2017 to 2019 in patients with Non-Hodgkin Lymphoma diagnosed with a biopsy. (3) Results: We included 39 patients who were newly diagnosed, untreated, and without any autoimmune disease previously reported. Thirty patients had the presence of autoantibodies (antiphospholipid antibodies, anti-cytoplasmic neutrophils antibodies, antinuclear antibodies), and nine were without autoantibodies. There were no statistical differences among groups regarding clinical, demographic, staging, and prognosis characteristics. Also, there were no differences in the outcomes of the patients after finishing chemotherapy and one year after initiating treatment. (4) Conclusions: Further investigations must be conducted regarding an extended panel of autoantibodies because the panel of autoantibodies in this study did not show a relationship between the presence and the clinical outcome of the patients.

Myxozoan survey of thicklip grey mullet Chelon labrosus reinforces successful radiation of Myxobolus in mugiliform hosts.

A myxozoan survey was performed on specimens of thicklip grey mullet Chelon labrosus (Risso) captured from the Douro River estuary, northern Portugal. Eleven new species, all belonging to the genus Myxobolus Bütschli, 1882 (M. abdominalis n. sp., M. aestuarium n. sp., M. caudalis n. sp., M. chelonari n. sp., M. cucurbitiformis n. sp., M. douroensis n. sp., M. intestinicola n. sp., M. invictus n. sp., M. labicola n. sp., M. peritonaei n. sp., and M. pinnula n. sp.) are described based on microscopic and molecular data, confirming the known high radiation of these myxozoans in mullets. Additionally, Myxobolus pupkoi Gupta et al., 2022 is reported for the first time from C. labrosus, bringing forth a novel case of morphological plasticity between geographic isolates. We consider that molecular-based comparisons are imperative for the description of mugiliform-infecting Myxobolus, with distance estimation further matching two of the novel Myxobolus spp. with sphaeractinomyxon types previously reported from another Portuguese estuary. This finding supports sphaeractinomyxon as specific life cycle counterparts of Myxobolus that infect mullets. Phylogenetic analyses of 18S rDNA retrieved a monophyletic clade of mugiliform-infecting myxobolids comprising well-supported lineages of species parasitizing mullets from the genera Chelon, Mugil, Crenimugil, and Planiliza. The existence of more than one Chelon- and Planiliza-infecting lineage reveals that myxobolids parasitized members of these genera multiple times during their evolution. Lastly, the elevated number of unmatched sphaeractinomyxon sequences included in the Chelon-infecting lineages clearly shows that Myxobolus diversity hosted by this genus remains underrated.

Title: Un inventaire des myxozoaires du mulet lippu Chelon labrosus confirme le rayonnement réussi de Myxobolus chez les hôtes mugiliformes. Abstract: Un inventaire des myxozoaires a été réalisé sur des spécimens de mulets lippus Chelon labrosus (Risso) capturés dans l'estuaire du fleuve Douro, au nord du Portugal. Onze nouvelles espèces, toutes appartenant au genre Myxobolus Bütschli, 1882 (M. abdominalis n. sp., M. aestuarium n. sp., M. caudalis n. sp., M. chelonari n. sp., M. cucurbitiformis n. sp., M. douroensis n. sp., M. intestinicola n. sp., M. invictus n. sp., M. labicola n. sp., M. peritonaei n. sp. et M. pinnula n. sp.) sont décrites sur la base de données microscopiques et moléculaires, confirmant le rayonnement connu de ces myxozoaires chez les mulets. De plus, Myxobolus pupkoi Gupta et al., 2022 est signalé pour la première fois chez C. labrosus, démontrant un nouveau cas de plasticité morphologique entre des isolats géographiques. Nous considérons que les comparaisons moléculaires sont impératives pour la description des Myxobolus infectant les mugiliformes, l'estimation de la distance correspondant en outre à deux des nouveaux Myxobolus spp. avec des types de sphaeractinomyxons précédemment signalés dans un autre estuaire portugais. Cette découverte soutient les sphaeractinomyxons en tant que contreparties spécifiques du cycle de vie de Myxobolus qui infectent les mulets. Les analyses phylogénétiques de l'ADNr 18S ont montré un clade monophylétique de Myxobolidae infectant les mugiliformes, comprenant des lignées robustes d'espèces parasitant les mulets des genres Chelon, Mugil, Crenimugil et Planiliza. L'existence de plusieurs lignées infectant Chelon et Planiliza révèle que les Myxobolidae ont parasité des membres de ces genres plusieurs fois au cours de leur évolution. Enfin, le nombre élevé de séquences de sphaeractinomyxons non appariées incluses dans les lignées infectant Chelon montre clairement que la diversité de Myxobolus hébergée par ce genre reste sous-estimée.

Avaliação da qualidade de vida relacionada à saúde em um programa de reabilitação para pessoas com deficiência visual / Health-related quality of life assessment in a rehabilitation program for people with visual impairment

RESUMO Objetivo: Aplicar um protocolo para avaliar a qualidade de vida relacionada à saúde de participantes de um programa de reabilitação para pessoas com deficiência visual de um instituto de referência. Métodos: Foi realizado um estudo transversal com 60 adultos com deficiência visual participantes de um programa de reabilitação para pessoas com deficiência visual de um instituto de referência no Rio de Janeiro. O protocolo de pesquisa consistiu em um questionário com dados pessoais, sociais, demográficos e informações clínicas; no European Quality of Life 5 Dimensions 3 Level Version para medição genérica de qualidade de vida relacionada à saúde; no Patient Health Questionnaire-2 para rastrear a depressão e no Visual Function Questionnaire 25 para avaliar a qualidade de vida relacionada à saúde específica da função visual. A principal variável independente analisada foi o tempo de exposição ao programa. Modelos de regressão linear foram utilizados para investigar a relação entre o tempo no programa e a qualidade de vida relacionada à saúde do Visual Function Questionnaire 25 e do European Quality of Life 5 Dimensions 3 Level Version. Resultados: A maioria dos participantes (73%) possuía deficiência visual adquirida; 68% tinham menos de 60 anos e 53% perderam a visão há mais de 10 anos. A condição visual autorreferida mais comum foi cegueira em ambos os olhos (48%) e 42% frequentavam o programa há mais de 3 anos. A mediana do índice de utilidade do European Quality of Life 5 Dimensions 3 Level Version foi de 0,75. O instrumento específico para rastreamento de depressão, o Patient Health Questionnaire, identificou proporção de 27% de participantes positivos. O Visual Function Questionnaire 25 apresentou escores abaixo de 50 (escala de zero a cem) nos subdomínios visão geral, atividades de perto e atividades à distância. As medianas de dor ocular e aspectos sociais do Visual Function Questionnaire 25 foram significativamente menores entre aqueles que realizavam tratamento psiquiátrico. O tempo de reabilitação foi independentemente associado a melhores escores dos subdomínios saúde mental e atividades da vida diária. Conclusão: O protocolo demonstrou aplicabilidade para a avaliação de qualidade de vida relacionada à saúde em pessoas com deficiência visual, permitindo concluir que o maior tempo no programa de reabilitação foi associado a maiores escores de qualidade de vida.

ABSTRACT Purpose: This study aimed to implement a health-related quality of life (HRQoL) assessment protocol to measure the consequences of a rehabilitation program for visual impaired people at a leading reference institute in Brazil. Methods: A cross-sectional study was conducted with 60 visual impaired adults enrolled in a Rehabilitation Program of the Instituto Benjamin Constant. The research protocol consisted of a questionnaire with personal data, social, demographic, and clinical information; the EQ-5D-3L instrument for generic HRQoL measurement; the Patient Health Questionnaire-2 (PHQ- 2) to screen for depression, and the 25-Item National Eye Institute Visual Function Questionnaire (NEI VFQ-25) to assess specific HRQoL of visual function. The main independent variable analyzed was the exposure time to the rehabilitation. Linear regression models were used to investigate the relationship between rehabilitation time and HRQoL of the NEI VFQ-25 and EQ 5D-3L instruments. Results: Most participants (73%) have acquired visual impairment, 68% are under 60 years old, 53% lost their vision more than 10 years ago, the most common self-reported visual condition (48%) was blindness in both eyes and 42% are in the Rehabilitation Program for more than 3 years. The median HRQoL utility index for EQ 5D-3L was 0.75. The specific instrument for screening for depression, PHQ-2, identified 27% of participants above the cut-off point. The NEI VFQ-25 instrument showed scores below 50 (scale from 0 to 100) in subdomains: "general vision", "near activities" and "distance activities". The medians of "ocular pain" and "social aspects" of the VFQ-25 were significantly lower among those who have undergone psychiatric treatment. "Rehabilitation time" was independently associated with better scores of "mental health" and "role difficulties" subdomains. Conclusion: The protocol showed applicability for the assessment of HRQoL, allowing the conclusion that longer time in the rehabilitation program was associated with higher quality of life scores.

Overcoming the Impact of Hypoxia in Driving Radiotherapy Resistance in Head and Neck Squamous Cell Carcinoma.

Hypoxia is very common in most solid tumours and is a driving force for malignant progression as well as radiotherapy and chemotherapy resistance. Incidences of head and neck squamous cell carcinoma (HNSCC) have increased in the last decade and radiotherapy is a major therapeutic technique utilised in the treatment of the tumours. However, effectiveness of radiotherapy is hindered by resistance mechanisms and most notably by hypoxia, leading to poor patient prognosis of HNSCC patients. The phenomenon of hypoxia-induced radioresistance was identified nearly half a century ago, yet despite this, little progress has been made in overcoming the physical lack of oxygen. Therefore, a more detailed understanding of the molecular mechanisms of hypoxia and the underpinning radiobiological response of tumours to this phenotype is much needed. In this review, we will provide an up-to-date overview of how hypoxia alters molecular and cellular processes contributing to radioresistance, particularly in the context of HNSCC, and what strategies have and could be explored to overcome hypoxia-induced radioresistance.

Antiphospholipid Antibodies and Lipids in Hematological Malignancies.

One of the main groups of lipids is phospholipids, which are mainly involved in forming cell membranes. Neoplastic processes such as cell replication have increased lipid synthesis, making tumor cells dependent on this synthesis to maintain their requirements. Antiphospholipid antibodies attack phospholipids in the cell membranes. Three main types of antiphospholipid antibodies are recognized: anti-ß2 glycoprotein I (anti-ß2GP-I), anticardiolipin (aCL), and lupus anticoagulant (LA). These types of antibodies have been proven to be present in hematological neoplasms, particularly in LH and NHL. This review on antiphospholipid antibodies in hematological neoplasms describes their clinical relationship as future implications at the prognostic level for survival and even treatment.

Regulation of chromatin accessibility by hypoxia and HIF.

Reduced oxygen availability (hypoxia) can act as a signalling cue in physiological processes such as development, but also in pathological conditions such as cancer or ischaemic disease. As such, understanding how cells and organisms respond to hypoxia is of great importance. The family of transcription factors called Hypoxia Inducible Factors (HIFs) co-ordinate a transcriptional programme required for survival and adaptation to hypoxia. However, the effects of HIF on chromatin accessibility are currently unclear. Here, using genome wide mapping of chromatin accessibility via ATAC-seq, we find hypoxia induces loci specific changes in chromatin accessibility are enriched at a subset hypoxia transcriptionally responsive genes, agreeing with previous data using other models. We show for the first time that hypoxia inducible changes in chromatin accessibility across the genome are predominantly HIF dependent, rapidly reversible upon reoxygenation and partially mimicked by HIF-α stabilisation independent of molecular dioxygenase inhibition. This work demonstrates that HIF is central to chromatin accessibility alterations in hypoxia, and has implications for our understanding of gene expression regulation by hypoxia and HIF.

Hemagglutination in gill capillaries of sheepshead, Archosargus probatocephalus (Perciformes: Sparidae), infected by a myxosporidean.

During a survey Myxozoa, four specimens of the sheepshead (18 ± 1.5 cm and 59 ± 2.5 g) (Archosargus probatocephalus) were collected in the Ipioquinha river (Maceió/AL). Transmission electron microscopy observations revealed erythrocyte agglutinations in gill capillaries located near spherical cysts containing myxospores of the genus Henneguya. This hemagglutination partially or totally obstructed the gill capillaries. Erythrocytes occurred in close adherence to each other, with a closed intercellular space. A few lysed erythrocytes were observed among agglutinated cells. The reduced lumen of the capillaries was partially filled with amorphous dense homogenous material adhering to the erythrocytes. In addition, heterogeneous masses of irregular lower electron density were observed in the reduced channel of the capillary. The agglutinated erythrocytes appeared dense and homogenous, lacking cytoplasmic organelles. The nuclei had the appearance of normal condensed chromatin masses, generally without visible nucleoli. This occurrence of hemagglutination only in the capillaries located in close proximity to the developing myxozoan cysts suggests that parasite development may be a factor triggering erythrocyte agglutination. This is supported by previous experimental studies that showed a probable correlation between parasitic infections and hemagglutination. Nonetheless, further studies are necessary in order to better understand the physicochemical processes involved in this phenomenon.

Macrophage Resistance to Ionizing Radiation Exposure Is Accompanied by Decreased Cathepsin D and Increased Transferrin Receptor 1 Expression.

PURPOSE: To identify a molecular signature of macrophages exposed to clinically relevant ionizing radiation (IR) doses, mirroring radiotherapy sessions. METHODS: Human monocyte-derived macrophages were exposed to 2 Gy/ fraction/ day for 5 days, mimicking one week of cancer patient's radiotherapy. Protein expression profile by proteomics was performed. RESULTS: A gene ontology analysis revealed that radiation-induced protein changes are associated with metabolic alterations, which were further supported by a reduction of both cellular ATP levels and glucose uptake. Most of the radiation-induced deregulated targets exhibited a decreased expression, as was the case of cathepsin D, a lysosomal protease associated with cell death, which was validated by Western blot. We also found that irradiated macrophages exhibited an increased expression of the transferrin receptor 1 (TfR1), which is responsible for the uptake of transferrin-bound iron. TfR1 upregulation was also found in tumor-associated mouse macrophages upon tumor irradiation. In vitro irradiated macrophages also presented a trend for increased divalent metal transporter 1 (DMT1), which transports iron from the endosome to the cytosol, and a significant increase in iron release. CONCLUSIONS: Irradiated macrophages present lower ATP levels and glucose uptake, and exhibit decreased cathepsin D expression, while increasing TfR1 expression and altering iron metabolism.

Hemagglutination in gill capillaries of sheepshead, Archosargus probatocephalus (Perciformes: Sparidae), infected by a myxosporidean / Hemaglutinação em capilares branquiais de sargo-de-dente, Archosargus probatocephalus (Perciformes: Sparidae) infectado por um mixosporídeo

Abstract During a survey Myxozoa, four specimens of the sheepshead (18 ± 1.5 cm and 59 ± 2.5 g) (Archosargus probatocephalus) were collected in the Ipioquinha river (Maceió/AL). Transmission electron microscopy observations revealed erythrocyte agglutinations in gill capillaries located near spherical cysts containing myxospores of the genus Henneguya. This hemagglutination partially or totally obstructed the gill capillaries. Erythrocytes occurred in close adherence to each other, with a closed intercellular space. A few lysed erythrocytes were observed among agglutinated cells. The reduced lumen of the capillaries was partially filled with amorphous dense homogenous material adhering to the erythrocytes. In addition, heterogeneous masses of irregular lower electron density were observed in the reduced channel of the capillary. The agglutinated erythrocytes appeared dense and homogenous, lacking cytoplasmic organelles. The nuclei had the appearance of normal condensed chromatin masses, generally without visible nucleoli. This occurrence of hemagglutination only in the capillaries located in close proximity to the developing myxozoan cysts suggests that parasite development may be a factor triggering erythrocyte agglutination. This is supported by previous experimental studies that showed a probable correlation between parasitic infections and hemagglutination. Nonetheless, further studies are necessary in order to better understand the physicochemical processes involved in this phenomenon.

Resumo Durante pesquisa de mixozoários foram coletados quatro espécimes do peixes sargo-de-dente (18 ± 1.5 cm e 59 ± 2.5 g) (Archosargus probatocephalus), no rio Ipioquinha (Maceió/AL). Observações por microscopia eletrônica de transmissão revelaram aglutinação de eritrócitos em capilares branquiais localizados próximos a cistos esféricos, contendo mixosporos do gênero Henneguya. Essa hemaglutinação obstruiu parcial ou totalmente os capilares branquiais. Os eritrócitos apareceram em forte aderência entre si, com espaço intercelular fechado. Foram observados poucos eritrócitos lisados entre as células aglutinadas. O lúmen reduzido dos capilares foi parcialmente preenchido com material homogêneo denso amorfo aderido aos eritrócitos, além de massas livres heterogêneas de densidade eletrônica baixa e irregular observadas no canal reduzido dos capilares. Os eritrócitos aglutinados pareciam densos e homogêneos, sem organelas citoplasmáticas. Os núcleos apareceram como massas normais de cromatina condensada, geralmente sem nucléolos visíveis. A ocorrência de hemaglutinação apenas nos capilares, localizados nas proximidades dos cistos mixozoários, sugere que o desenvolvimento parasitário pode ser um fator desencadeante da aglutinação eritrocitária. Isso é corroborado por estudos experimentais anteriores que mostraram uma provável correlação entre infecções parasitárias e hemaglutinação. No entanto, novos estudos são necessários para melhor compreender os processos físico-químicos envolvidos neste fenômeno.

Optimization and Validation of an In Vitro Standardized Glycogen Phosphorylase Activity Assay.

Glycogen phosphorylase (GP) is a key enzyme in the glycogenolysis pathway and a potential therapeutic target in the management of type 2 diabetes. It catalyzes a reversible reaction: the release of the terminal glucosyl residue from glycogen as glucose 1-phosphate; or the transfer of glucose from glucose 1-phosphate to glycogen. A colorimetric method to follow in vitro the activity of GP with usefulness in structure-activity relationship studies and high-throughput screening capability is herein described. The obtained results allowed the choice of the optimal concentration of enzyme of 0.38 U/mL, 0.25 mM glucose 1-phosphate, 0.25 mg/mL glycogen, and temperature of 37 °C. Three known GP inhibitors, CP-91149, a synthetic inhibitor, caffeine, an alkaloid, and ellagic acid, a polyphenol, were used to validate the method, CP-91149 being the most active inhibitor. The effect of glucose on the IC50 value of CP-91149 was also investigated, which decreased when the concentration of glucose increased. The assay parameters for a high-throughput screening method for discovery of new potential GP inhibitors were optimized and standardized, which is desirable for the reproducibility and comparison of results in the literature. The optimized method can be applied to the study of a panel of synthetic and/or natural compounds, such as polyphenols.