Horismenus camobiensis (Hym Eulophidae), a new hyperparasitoid of Cotesia invirae (Hym Braconidae) in Opsiphanes invirae (Lep Nymphalidae) pupae / Horismenus camobiensis (Hym Eulophidae), um novo hiperparasitóide de Cotesia invirae (Hym Braconidae) em pupas de Opsiphanes invirae (Lep Nymphalidae)

Abstract Horismenus camobiensis sp. nov. (Hymenoptera: Eulophidae), is described based on morphological, molecular and ecological data; this new species of chalcid wasp acts as hyperparasitoid of Opsiphanis invirae (Hübner, 1818) (Lepidoptera: Nymphalidae) in its parasitoid Cotesia invirae Salgado-Neto and Whitfield, 2019 (Hymenoptera: Braconidae). Diagnoses with morphological and molecular characters and illustrations are provided.

Resumo Horismenus camobiensis sp. nov. (Hymenoptera: Eulophidae) é descrita com base em dados morfológicos, moleculares e ecológicos; esta nova espécie Chalcididae atua como hiperparasitoide de Opsiphanis invirae (Hübner, 1818) (Lepidoptera: Nymphalidae) em pupas de seu parasitoide Cotesia invirae Salgado-Neto and Whitfield, 2019 (Hymenoptera: Braconidae). Caracteres diagnósticos morfológicos e moleculares e ilustrações de H. camobiensis são fornecidos.

Horismenus camobiensis (Hym.: Eulophidae), a new hyperparasitoid of Cotesia invirae (Hym.: Braconidae) in Opsiphanes invirae (Lep.: Nymphalidae) pupae

Abstract Horismenus camobiensis sp. nov. (Hymenoptera: Eulophidae), is described based on morphological, molecular and ecological data; this new species of chalcid wasp acts as hyperparasitoid of Opsiphanis invirae (Hübner, 1818) (Lepidoptera: Nymphalidae) in its parasitoid Cotesia invirae Salgado-Neto and Whitfield, 2019 (Hymenoptera: Braconidae). Diagnoses with morphological and molecular characters and illustrations are provided.

Resumo Horismenus camobiensis sp. nov. (Hymenoptera: Eulophidae) é descrita com base em dados morfológicos, moleculares e ecológicos; esta nova espécie Chalcididae atua como hiperparasitoide de Opsiphanis invirae (Hübner, 1818) (Lepidoptera: Nymphalidae) em pupas de seu parasitoide Cotesia invirae Salgado-Neto and Whitfield, 2019 (Hymenoptera: Braconidae). Caracteres diagnósticos morfológicos e moleculares e ilustrações de H. camobiensis são fornecidos.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul – Brazil / Primeira lista de parasitoides em moscas de importância médica, veterinária e forense no Rio Grande do Sul - Brasil

Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42' S; 52º 18' W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42' S; 52º 18' O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul Brazil

Abstract Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42 S; 52º 18 W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Resumo Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42 S; 52º 18 O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

First list of parasitoids in flies of medical, veterinary, and forensic importance in Rio Grande do Sul - Brazil / Primeira lista de parasitoides em moscas de importância médica, veterinária e forense no Rio Grande do Sul - Brasil

Necrophagous flies are of great importance for human and animal health. In places where their development occurs, parasitoids can be used as a tool to control these dipterans. In Brazil, the fauna of these parasitoids has been investigated in some regions. However, in Rio Grande do Sul, it is known the occurrence of only one species. Thus, this study aimed to create the first list of parasitoids in flies of medical and veterinary importance in Southern Brazil. Collections took place in the municipality of Pelotas (31º 42' S; 52º 18' W). Three replicates consisting of a 500 g bovine liver chunk placed in a tray were exposed to open air for 20 days. Then pupae were individualized and observed until their emergence. We identified 4,882 adult flies of Calliphoridae, Fanniidae, Muscidae, and Sarcophagidae. 4,040 parasitoids emerged, belonging to eight species, of which Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis, and Tachinephagus zealandicus are new records in Rio Grande do Sul state. Also, three undescribed species of the genus Aphaereta were collected. In spite of being the first attempt to inventory the fauna of hymenopteran parasitoids, this study may help in the development of management programs of these dipterans in the region.

Moscas necrófagas possuem grande importância para a sanidade humana e animal. Onde estas se desenvolvem ocorrem parasitoides que podem ser utilizados como ferramentas para o controle desses dípteros. No Brasil, a fauna desses parasitoides vem sendo investigada em algumas regiões e no Rio Grande do Sul é conhecida a ocorrência de apenas uma espécie. Portanto, o objetivo deste estudo foi criar a primeira lista de espécies de parasitoides de moscas de importância médica e veterinária ocorrentes no extremo sul do Brasil. As coletas foram realizadas no município de Pelotas (31º 42' S; 52º 18' O). Foram montadas três réplicas de bandejas contendo, cada uma, 500 g de fígado bovino, expostas ao ar livre durante 20 dias. Na sequência, as pupas foram individualizadas e observadas até a emergência. Foram identificados 4.882 adultos pertencentes às famílias Calliphoridae, Fanniidae, Muscidae e Sarcophagidae. Emergiram 4.040 parasitoides, pertencentes a oito espécies, das quais Nasonia vitripennis, Spalangia cameroni, Spalangia chontalensis e Tachinephagus zealandicus são novas ocorrências para o Rio Grande do Sul. Foram coletadas três espécies ainda não descritas do gênero Aphaereta. Apesar de ainda ser um primeiro esforço de inventariar a fauna de himenópteros parasitoides, este estudo pode auxiliar no desenvolvimento de programas de manejos desses dípteros na região.

CNS-3 status remains an independent adverse prognosis factor in children with acute lymphoblastic leukemia (ALL) treated without cranial irradiation: Results of EORTC Children Leukemia Group study 58951.

AIM: To evaluate the prognostic significance of initial central nervous system (CNS) involvement of children with acute lymphoblastic leukemia (ALL) enrolled in the EORTC 58951 trial. PATIENTS AND METHODS: From 1998 to 2008, 1930 ALL patients were included in the randomized EORTC 58951 trial. Overall treatment intensity was adjusted according to known prognostic factors including the level of minimal residual disease after induction treatment. CNS-directed therapy comprised four to 11 courses of i.v. methotrexate (5g/m2), and 10 to 19 intrathecal chemotherapy injections, depending on risk group and CNS status. Cranial irradiation was omitted for all patients. RESULTS: The overall 8-year event-free survival (EFS) and overall survival (OS) rates were 81.3% and 88.1%, respectively. In the CNS-1, TPL+, CNS-2, and CNS-3 groups, the 8-year EFS rates were 82.1%, 77.1%, 78.3%, and 57.4%, respectively. Multivariable analysis indicated that initial CNS-3 status, but not CNS-2 or TLP+, was an independent adverse predictor of outcome. The 8-year incidence of isolated CNS relapse was 1.7% and of isolated or combined CNS relapse it was 3.7%. NCI high-risk group, male sex, CNS-2 and CNS-3 status were independent predictors for a higher incidence of any CNS relapse. CONCLUSIONS: CNS-3 status remains associated with poor prognosis and requires intensification of both systemic and CNS-directed therapy. This trial was registered at https://clinicaltrials.gov/under/NCT00003728.

Microencapsulated and uncoated butyric acid as alternative additives to the regeneration of intestinal mucosa in broilers challenged with Eimeria spp.

1. The effect of microencapsulated and uncoated butyric acid as an alternative to antibiotics on performance, intestinal morphology and regeneration of intestinal mucosa was studied in birds experimentally infected with Eimeria spp. 1 to 42 d-old.2. A total of 1,320 male Cobb® broiler chicks were allocated to one of five treatments in a completely randomised design, comprising a negative control, uncoated butyric acid (UA), microencapsulated butyric acid (MA), combined U + M butyric acid and a positive control (antibiotic+anticoccidial) in six replications. At 16 d-old, the birds were inoculated orally with 0.5 ml of a solution containing an Eimeria spp. pool.3. At 21 d of age, the birds receiving butyric acid alone had higher body weight gain (BWG) and feed intake (FI) compared to those supplemented with the blend of acids. For the total rearing period, in all variables, the positive control performed best (P < 0.001).4. At 14 d of age, birds that received diets containing UA had a deeper crypt depth in the jejunum than those fed diets containing microencapsulated acid (P = 0.0194). At 21 d of age, the birds fed the acids had higher villi (P = 0.0058) in the duodenum, compared to the negative control group.5. Supplementation with microencapsulated acid contributed to the intestinal health and recovery of post-challenge birds, but did not result in improvements in performance.

Deregulated miRNAs in osteoporosis: effects in bone metastasis.

Starting from their role exerted on osteoblast and osteoclast differentiation and activity pathways, microRNAs (miRNAs) have been recently identified as regulators of different processes in bone homeostasis. For this purpose, in a recent review, we highlighted, as deregulated miRNAs could be involved in different bone diseases such as osteoporosis. In addition, recent studies supported the concept that osteoporosis-induced bone alterations might offer a receptive site for cancer cells to form bone metastases, However, to date, no data on specific-shared miRNAs between osteoporosis and bone metastases have been considered and described to clarify the evidence of this link. The main goal of this review is to underline as deregulated miRNAs in osteoporosis may have specific roles in the development of bone metastases. The review showed that several circulating osteoporotic miRNAs could facilitate tumor progression and bone-metastasis formation in several tumor types, i.e., breast cancer, prostate cancer, non-small-cell lung cancer, esophageal squamous cell carcinoma, and multiple myeloma. In detail, serum up-regulation of pro-osteoporotic miRNAs, as well as serum down-regulation of anti-osteoporotic miRNAs are common features of all these tumors and are able to promote bone metastasis. These results are of key importance and could help researcher and clinicians to establish new therapeutic strategies connected with deregulation of circulating miRNAs and able to interfere with pathogenic processes of osteoporosis, tumor progressions, and bone-metastasis formation.

The effects of xanthan gum on equine sperm quality during cooling storage / Efeitos de xantam gum em qualidade de esperma equino durante armazenamento resfriado

This study was designed to evaluate the possible benefits of adding xanthan gum to a standard extender for equine through in vitro analyzes of sperm quality. Semen was collected four times from five different stallions (n= 20 samples) and subjected to cooled storage under different conditions: control (only standard extender) and three different concentrations of xanthan gum (0.01%, 0.12%, and 0.25%) supplemented to the extenders. Sperm parameters, such as motility, mitochondrial functionality, and membrane, acrosome, and DNA integrity were measured after 0h, 24h, 48h, and 72h of sperm storage at 5ºC. Our observations indicated that sperm motility declined with longer cooling period with the 0.25% xanthan gum supplementation group compared with the control group. Other parameters, such as mitochondrial functionality and membrane and acrosome integrity also declined for all treatments during storage; however, no differences were observed between xanthan gum and control groups. DNA integrity did not significantly change during the storage. In conclusion, the addition of xanthan gum to equine semen extender is not harmful to the sperm structure, despite reducing the sperm motility.(AU)

Esse estudo foi desenvolvido para avaliar os possíveis benefícios de acrescentar xanthan gum a um extensor padrão através de analises in vitro de qualidade de esperma. Semen foi coletado quatro vezes de cinco garanhões diferentes (n = 20 amostras) e submetido a armazenamen to resfriado em diferentes condições: controle (apenas extensor padrão) e três diferentes concentrações de xanthan gum (0,01%, 0,12% e 0,25%) suplementado aos extensores. Parâmetros dos espermatozoides, como mobilidade, funcionamento mitocondrial e integridade de membranas, acrossomos e DNA forma medidos após 0h, 24h, 48h e 72h de armazenamento a 5oC. Nossas observações indicaram que motilidade reduziu com armazenamento resfriado prolongado no grupo de 0,25% de suplementação de xanthan gum comparado ao grupo controle. Outros parâmetros, como funcionalidade mitocondrial e integridade de membrana e acrossomos também reduziu em todos os tratamentos durante o armazenamento, no entanto não foram detectadas diferenças significativas entre grupos tratados e grupo controle. Integridade de DNA não mudou significativamente durante armazenamento. Em conclusão, a adição de xanthan gum a extensor de sêmen equino não é danosa à estrutura do espermatozoide apesar de reduzir motilidade.(AU)

Engineered exosomes: A new promise for the management of musculoskeletal diseases.

BACKGROUND: Exosomes are nanovesicles actively secreted by potentially all cell types, including tumour cells, with the primary role of extracellular systemic communication mediators, both at autocrine and paracrine levels, at short and long distances. Recently, different studies have used exosomes as a delivery system for a plethora of different molecules, such as drugs, microRNAs and proteins. This has been made possible thanks to the simplicity in exosomes engineering, their great stability and versatility for applications in oncology as well as in regenerative medicine. SCOPE OF REVIEW: The aim of this review is to provide information on the state-of-the-art and possible applications of engineered exosomes, both for cargo and specific cell-targeting, in different pathologies related to the musculoskeletal system. MAJOR CONCLUSIONS: The use of exosomes as therapeutic agents is rapidly evolving, different studies explore drug delivery with exosomes using different molecules, showing an enormous potential in various research fields such as oncology and regenerative medicine. GENERAL SIGNIFICANCE: However, despite the significant progress made by the different studies carried out, currently, the use of exosomes is not a therapeutic reality for the considerable difficulties to overcome.

EGFR amplification and expression in oral squamous cell carcinoma in young adults.

The aim of this study was to investigate epidermal growth factor receptor (EGFR) gene alterations in two groups of patients with oral squamous cell carcinoma (OSCC) (a test group of subjects aged ≤40 years and a control group of subjects aged ≥50 years) and to associate the results with EGFR immunostaining, clinicopathological features, and the prognosis. Sixty cases of OSCC were selected (test group, n=21; control group, n=39). The tissue microarray technique was applied to ensure the uniformity of results. Gene amplification was analyzed by fluorescence in situ hybridization (FISH), and immunohistochemical staining for EGFR was analyzed using an automated imaging system. EGFR amplification was higher in the test group than in the control group (P=0.018) and was associated with advanced clinical stage (P=0.013), regardless of age. Patients with EGFR overexpression had worse survival rates, as did patients who had T3-T4 tumours and positive margins. EGFR overexpression has a negative impact on disease progression. Despite the higher amplification of EGFR in young adults, it does not significantly impact the survival rates of affected patients.

Gene therapy for chondral and osteochondral regeneration: is the future now?

Gene therapy might represent a promising strategy for chondral and osteochondral defects repair by balancing the management of temporary joint mechanical incompetence with altered metabolic and inflammatory homeostasis. This review analysed preclinical and clinical studies on gene therapy for the repair of articular cartilage defects performed over the last 10 years, focussing on expression vectors (non-viral and viral), type of genes delivered and gene therapy procedures (direct or indirect). Plasmids (non-viral expression vectors) and adenovirus (viral vectors) were the most employed vectors in preclinical studies. Genes delivered encoded mainly for growth factors, followed by transcription factors, anti-inflammatory cytokines and, less frequently, by cell signalling proteins, matrix proteins and receptors. Direct injection of the expression vector was used less than indirect injection of cells, with or without scaffolds, transduced with genes of interest and then implanted into the lesion site. Clinical trials (phases I, II or III) on safety, biological activity, efficacy, toxicity or bio-distribution employed adenovirus viral vectors to deliver growth factors or anti-inflammatory cytokines, for the treatment of osteoarthritis or degenerative arthritis, and tumour necrosis factor receptor or interferon for the treatment of inflammatory arthritis.

Toxicity of the amphetamine metabolites 4-hydroxyamphetamine and 4-hydroxynorephedrine in human dopaminergic differentiated SH-SY5Y cells.

Amphetamine (AMPH) is a psychostimulant used worldwide by millions of patients in the clinical treatment of attention deficit hyperactivity disorder, narcolepsy or even obesity, and is also a drug of abuse. 4-Hydroxynorephedrine (4-OHNE) and 4-hydroxyamphetamine (4-OHAMPH) are two major metabolites known to persist in the brain longer than AMPH. The contribution of AMPH metabolites for its neurotoxicity is undetermined. We evaluated the toxicity of AMPH and its metabolites 4-OHNE and 4-OHAMPH, obtained by chemical synthesis, in human dopaminergic differentiated SH-SY5Y neurons. Cells were exposed to AMPH (concentration range 0-5mM) or 4-OHAMPH or 4-OHNE (concentration range 0-10mM) for 24 or 48h, and the viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) leakage assays. Results showed that for both AMPH and the metabolites a concentration-dependent toxicity was observed. The toxic concentration 50% (TC50) for AMPH and 4-OHNE following 24h exposure was circa 3.5mM and 8mM, respectively. For 4-OHAMPH the TC50 was not reached in the tested concentration range. N-acetyl cysteine, cycloheximide, l-carnitine, and methylphenidate were able to reduce cell death induced by AMPH TC50. Acridine orange/ethidium bromide staining showed evident signs of late apoptotic cells and necrotic cells following 24h exposure to AMPH 3.50mM. The 4-OHAMPH metabolite at 8.00mM originated few late apoptotic cells, whereas 4-OHNE at 8.00mM resulted in late apoptotic cells and necrotic cells, in a scenario similar to AMPH. In conclusion, the AMPH metabolite 4-OHNE is more toxic than 4-OHAMPH, nonetheless both are less toxic than the parent compound in vitro. The most toxic metabolite 4-OHNE has longer permanence in the brain, rendering likely its contribution for AMPH neurotoxicity.

Comparative analysis of three histologic grading methods for squamous cell carcinoma of the lip.

OBJECTIVES: The aim of this study was to evaluate three histologic grading methods for squamous cell carcinoma (SCC) of the lip, the conventional three-grade model proposed by the World Health Organization, tumor budding and depth of invasion (BD) model, and histologic risk assessment (HRA) model, and to correlate them with prognosis. MATERIALS AND METHODS: Fifty-three patients with lip SCC were evaluated. RESULTS: The mean age was 65 years, 69.8% of the participants were men, and 66.0% of the patients had early-stage tumors. Using the BD and conventional three-grade methods, 52.8% and 64.2% of the cases were graded as low risk, respectively. The HRA model graded 54.7% of the cases as medium risk. In the BD model, the higher histologic grade was associated with worse prognosis (P = 0.045). Overall survival at 5 years was 87.8%. Tumor size (T3 + T4) and lymph node involvement (N+) were associated with reduced overall survival and recurrence-free survival (RFS) (P = 0.002 and 0.005; 0.007 and 0.01, respectively). Surgical treatment combined with radiotherapy was associated with lower RFS (P = 0.03). CONCLUSIONS: High-grade lip SCC in advanced stages is associated with a poor prognosis. The BD model is a simple and effective tool for the prognostic evaluation of lip SCC.

A new bi-layered scaffold for osteochondral tissue regeneration: In vitro and in vivo preclinical investigations.

Current treatments for acute or degenerative chondral and osteochondral lesions are in need of improvement, as these types of injuries lead to disability and worsen the quality of life in a high percentage of patients. The aim of this study was to develop a new bi-layered scaffold for osteochondral tissue regeneration through a "biomimetic" and "bioinspired" approach. For chondral regeneration, the scaffold was realized with an organic compound (type I collagen), while for the regeneration of the subchondral layer, bioactive magnesium-doped hydroxyapatite (Mg/HA) crystals were co-precipitated with the organic component of the scaffold. The entire scaffold structure was stabilized with a cross-linking agent, highly reactive bis-epoxyde (1,4-butanediol diglycidyl ether - BDDGE 1wt%). The developed scaffold was then characterized for its physico-chemical characteristics. Its structure and adhesion strength between the integrated layers were investigated. At the same time, in vitro cell culture studies were carried out to examine the ability of chondral and bone scaffold layers to separately support adhesion, proliferation and differentiation of human mesenchymal stem cells (hMSCs) into chondrocytes and osteoblasts, respectively. Moreover, an in vivo study with nude mice, transplanted with osteochondral scaffolds plain or engineered with undifferentiated hMSCs, was also set up with 4 and 8-week time points. The results showed that chondral and bone scaffold layers represented biocompatible scaffolds able to sustain hMSCs attachment and proliferation. Moreover, the association of scaffold stimuli and differentiation medium, induced hMSCs chondrogenic and osteogenic differentiation and deposition of extracellular matrix (ECM). The ectopic implantation of the engineered osteochondral scaffolds indicated that hMSCs were able to colonize the osteochondral scaffold in depth. The scaffold appeared permissive to tissue growth and penetration, ensuring the diffusion of nutrients and oxygen, as also suggested by the presence of a neo-angiogenesis process, especially at 4weeks. Moreover, the in vivo results further confirmed the great potential of the scaffold in tissue engineering, as it was able to support the initial formation of new bone and chondral tissue, confirming the importance of combined and innovative strategies to improve the available therapeutic strategies for chondral and osteochondral regeneration.

Naphthoquinoxaline metabolite of mitoxantrone is less cardiotoxic than the parent compound and it can be a more cardiosafe drug in anticancer therapy.

Mitoxantrone (MTX) is an antineoplastic agent used to treat several types of cancers and on multiple sclerosis, which shows a high incidence of cardiotoxicity. Still, the underlying mechanisms of MTX cardiotoxicity are poorly understood and the potential toxicity of its metabolites scarcely investigated. Therefore, this work aimed to synthesize the MTX-naphthoquinoxaline metabolite (NAPHT) and to compare its cytotoxicity to the parent compound in 7-day differentiated H9c2 cells using pharmacological relevant concentrations (0.01-5 µM). MTX was more toxic in equivalent concentrations in all cytotoxicity tests performed [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction, neutral red uptake, and lactate dehydrogenase release assays] and times tested (24 and 48 h). Both MTX and NAPHT significantly decreased mitochondrial membrane potential in 7-day differentiated H9c2 cells after a 12-h incubation. However, energetic pathways were affected in a different manner after MTX or NAPHT incubation. ATP increased and lactate levels decreased after a 24-h incubation with MTX, whereas for the same incubation time and concentrations, NAPHT did not cause any significant effect. The increased activity of ATP synthase seems responsible for MTX-induced increases in ATP levels, as oligomycin (an inhibitor of ATP synthase) abrogated this effect on 5 µM MTX-incubated cells. 3-Methyladenine (an autophagy inhibitor) was the only molecule to give a partial protection against the cytotoxicity produced by MTX or NAPHT. To the best of our knowledge, this was the first broad study on NAPHT cardiotoxicity, and it revealed that the parent drug, MTX, caused a higher disruption in the energetic pathways in a cardiac model in vitro, whereas autophagy is involved in the toxicity of both compounds. In conclusion, NAPHT is claimed to largely contribute to MTX-anticancer properties; therefore, this metabolite should be regarded as a good option for a safer anticancer therapy since it is less cardiotoxic than MTX.

Direct laser writing of nanorough cell microbarriers on anatase/Si and graphite/Si.

The formation of hierarchical structures consisting of microstripe barriers decorated with nanorough ablated materials prepared by direct laser writing is described. Linear features of circa 25µm width and 12µm height are achieved on amorphous and crystalline titania and graphitic carbon films deposited on silicon. Ablated protrusions build up barriers decorated by nanoscale Si-film reconstructions, as indicated by EDX maps and micro-Raman spectroscopy. Wettability tests show a dramatic change in water contact angle, which leads to almost full wetting after irradiation, irrespective of the original film composition. Fluorescence microscopy images of human mesenchymal stem cells cultured on 1D and 2D structures demonstrate the short term biocompatibility of the ablated surfaces. It is shown that cells adhere, extend and polarize on feature edges, independently of the type of surface, thus suggesting that the created nanoroughness is at the origin of the antifouling behavior. In particular, irradiated anatase and graphite surfaces demonstrate an increased performance of crystalline films for the creation of cell guiding and trapping devices. The results suggest that such laser processing of films may serve as a time-and-cost-efficient method for the design of few-cells analytical surfaces.