Osteosarcoma After Total Knee Arthroplasty: A Case Report.

CASE: A 79-year-old woman presented with a periprosthetic fracture 8 years after a total knee arthroplasty (TKA). Radiographs demonstrated tibial implant loosening with severe osteolysis. A high-grade osteosarcoma around the prosthesis was diagnosed, and a supracondylar femoral amputation was performed. After 2 years, no complications have occurred. CONCLUSIONS: A malignant tumor around a TKA is extremely rare. Surgeons should remain vigilant with patients who present with rapidly progressive or very aggressive implant loosening with osteolysis. Owing to its complexity and potentially devastating prognosis, treatment should be guided by a specialist multidisciplinary team. Complex limb salvage procedures or amputation is usually required.

Expanding a precision medicine platform for malignant peripheral nerve sheath tumors: New patient-derived orthotopic xenografts, cell lines and tumor entities.

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive soft-tissue sarcomas with a poor survival rate, presenting either sporadically or in the context of neurofibromatosis type 1 (NF1). The histological diagnosis of MPNSTs can be challenging, with different tumors exhibiting great histological and marker expression overlap. This heterogeneity could be partly responsible for the observed disparity in treatment response due to the inherent diversity of the preclinical models used. For several years, our group has been generating a large patient-derived orthotopic xenograft (PDOX) MPNST platform for identifying new precision medicine treatments. Herein, we describe the expansion of this platform using six primary tumors clinically diagnosed as MPNSTs, from which we obtained six additional PDOX mouse models and three cell lines, thus generating three pairs of in vitro-in vivo models. We extensively characterized these tumors and derived preclinical models, including genomic, epigenomic, and histological analyses. Tumors were reclassified after these analyses: three remained as MPNSTs (two being classic MPNSTs), one was a melanoma, another was a neurotrophic tyrosine receptor kinase (NTRK)-rearranged spindle cell neoplasm, and, finally, the last was an unclassifiable tumor bearing neurofibromin-2 (NF2) inactivation, a neuroblastoma RAS viral oncogene homolog (NRAS) oncogenic mutation, and a SWI/SNF-related matrix-associated actin-dependent regulator of chromatin (SMARCA4) heterozygous truncated variant. New cell lines and PDOXs faithfully recapitulated histology, marker expression, and genomic characteristics of the primary tumors. The diversity in tumor identity and their specific associated genomic alterations impacted treatment responses obtained when we used the new cell lines for testing compounds against known altered pathways in MPNSTs. In summary, we present here an extension of our MPNST precision medicine platform, with new PDOXs and cell lines, including tumor entities confounded as MPNSTs in a real clinical scenario. This platform may constitute a useful tool for obtaining correct preclinical information to guide MPNST clinical trials.

Cost-effective fibrinolytic enzyme production by microalga Dunaliella tertiolecta using medium supplemented with corn steep liquor.

A fibrinolytic enzyme from the microalga Dunaliella tertiolecta was produced under mixotrophic conditions using different corn steep liquor (CSL) concentrations ( 0 ≤ CLS ≤ 0.75%), purified using a combination of salting out and ion-exchange chromatography, and then biochemical characterized. Cultivation of this microalga using 0.5% CSL led to the highest maximum cell concentration (1.960±0.010 mg L-1) and cell productivity (0.140g L-1 day-1), besides a high fibrinolytic activity of the extract obtained by the homogenization method (102 ±1 U mL-1). The enzyme extracted from the microalgal biomass was 5-fold purified with a 20% yield and was found to have a specific activity of 670 U mg-1. The enzyme, whose molecular weight determined by fibrin zymography was 10 kDa, was shown to be stable at pH 3.0-9.0 and up to 70°C with optimal pH and temperature values of 8.0 and 50°C, respectively. When compared to other fibrinolytic enzymes, this protease stood out for its high fibrinolytic activity, which was enhanced by Fe2+, inhibited by Zn2+, Cu2+, Mg2+, and Ca2+, and strongly inhibited by phenylmethylsulfonyl fluoride, suggesting that it belongs to the serine metalloprotease family. Moreover, thanks to its thermal stability, the enzyme may be easily preserved and activated under high-temperature conditions.

Histological response to radiotherapy is an early event in myxoid liposarcoma.

Compared to other sarcomas, myxoid liposarcoma (MLS) is exceptionally sensitive to radiation therapy, but the underlying mechanism remains unknown. The objective was to assess the tissue-based changes in MLS during and after neoadjuvant radiotherapy in 26 patients of the DOREMY trial. Morphological assessment was performed on biopsies pre-treatment, after 8 fractions, 16 factions, and after surgical resection and included percentage of viable tumor cells, hyalinization, necrosis, and fatty maturation. Furthermore, immunohistochemistry was performed for apoptosis (cleaved caspase-3), anti-apoptosis (Bcl-2), activity of mTOR signaling (phospho-S6), hypoxia (CAIX), proliferation (Ki67), inflammation (CD45 and CD68), and microvessel density (CD34 Chalkley count). A pronounced reduction in vital tumor cells was observed early with a drop to 32.5% (median) tumor cells (IQR 10-93.8%) after 8 fractions. This decreased further to 10% (IQR 5-30%) after 16 fractions and 7.5% (IQR 5-15%) in the surgical specimen. All but one patient had an excellent response with < 50% remaining tumor cells. Inversely, treatment response was mainly observed as hyalinization and less often as fatty maturation. Additionally, a decrease of inflammatory cells was noticed especially during the first eight fractions. Microvessel density remained stable over time. Immunohistochemical markers for apoptosis, anti-apoptosis, activity of mTOR signaling, proliferation, and hypoxia did not show any marked changes within the remaining tumor cells during and after radiotherapy. As a modest dose of neoadjuvant radiotherapy induces profound tissue changes in MLS, mainly during the first 8 fractions, current findings might suggest that in a carefully selected patient population further deintensification of radiotherapy might be explored.

KRT5+/p63+ Stem Cells Undergo Senescence in the Human Lung with Pathological Aging.

Adult lungs present high cellular plasticity against stress and injury, mobilizing stem/progenitor populations from conducting airways to maintain tissue homeostasis and gas exchange in alveolar spaces. With aging, pulmonary functional and structural deterioration occurs, mainly in pathological conditions, which is associated with impaired stem cell activity and increased senescence in mice. However, the impact of these processes underlying lung physiopathology in relation to aging has not been explored in humans. In this work, we analyzed stem cell (SOX2, p63, KRT5), senescence (p16INK4A, p21CIP, Lamin B1) and proliferative (Ki67) markers in lung samples from young and aged individuals, with and without pulmonary pathology. We identified a reduction in SOX2+ cells but not p63+ and KRT5+ basal cells in small airways with aging. In alveoli, we revealed the presence of triple SOX2+, p63+ and KRT5+ cells specifically in aged individuals diagnosed with pulmonary pathologies. Notably, p63+ and KRT5+ basal stem cells displayed colocalization with p16INK4A and p21CIP, as well as with low Lamin B1 staining in alveoli. Further studies revealed that senescence and proliferation markers were mutually exclusive in stem cells with a higher percentage colocalizing with senescence markers. These results provide new evidence of the activity of p63+/KRT5+ stem cells on human lung regeneration and point out that regeneration machinery in human lung is activated under stress due to aging, but fails to repair in pathological cases, as stem cells would likely become senescent.

New molecular entities of soft tissue and bone tumors.

PURPOSE OF REVIEW: The advances of molecular techniques have led to the refinement of the classification of mesenchymal tumors, leading to newly introduced entities in the recently published fifth edition of the WHO Classification of Soft Tissue and Bone Tumors, which are discussed in this review. RECENT FINDINGS: For the first time, entities are included of which the name refers to the underlying molecular alteration including round cell sarcoma with EWSR1 -non-ETS fusions, CIC -rearranged sarcoma, and sarcoma with BCOR genetic alteration. EWSR1-SMAD3 -positive fibroblastic tumor and NTRK -rearranged spindle cell neoplasm are provisionally included as 'emerging' entities based on the underlying molecular alteration, though the entity still needs to be better defined. Other newly recognized entities are not named after their molecular change, but the molecular alteration helped to delineate them from others: atypical spindle cell/pleomorphic lipomatous tumor, anastomosing hemangioma, angiofibroma of soft tissue, myxoid pleomorphic liposarcoma, and poorly differentiated chordoma. SUMMARY: Classification of mesenchymal tumors is increasingly based on the underlying molecular changes, although this cannot be interpreted separately from clinical, morphological, and immunohistochemical characteristics.

Accuracy of fine-needle aspiration of lymph nodes: A cancer center's experience.

INTRODUCTION: Lymph node fine needle aspiration (LN-FNA) is a minimally invasive method of evaluating lymphadenopathy. Nonetheless, its use is not widely accepted due to the lack of guidelines and a cytopathological categorisation that directly relates to management. We report our experience with LN FNA at a large Cancer Center in Latin America. METHODS: We retrospectively collected cytological cases of lymph node FNA from the department of pathology at AC Camargo Cancer Center performed over a 2-year period. Data extracted included LN location, age, sex and final cytological diagnosis. Patients that had undergone neoadjuvant chemotherapy and/or cases for which the surgery specimen location was not clearly reported were excluded. For those cases with surgical reports, risk of malignancy was calculated for each diagnostic category, along with overall performance of cytology. False positive cases were reviewed to assess any possible misinterpretation or sampling errors. RESULTS: A total of 1730 LN-FNA were distributed as follows: 62 (3.5%) non-diagnostic (ND); 1123 (64.9%) negative (NEG), 19 (1.1%) atypical (ATY), 53 (3.1%) suspicious for malignancy (SUS), and 473 (27.3%) positive (POS). Surgical reports were available for 560 cases (32.4%). Risk of malignancy (ROM) for each category was 33.3% for ND, 29.9% for NEG, 25% for ATY, 74.2% for SUS and 99.6% for POS. Overall sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) were 78.5%, 99.4%, 70.2% and 99.6%, respectively. CONCLUSION: Lymph node FNA is a very specific and accurate exam, which is reliable in the detection of lymph node metastasis and other causes of lymphadenopathy.

Produções sobre neuropsicologia e suas contribuições para a (DES) patologização da educação / Productions on Neuropsychology and their contributions to the (de)pathologization of education

INTRODUÇÃO: Esse artigo lança como objetivo geral analisar a vinculação temática entre Neuropsicologia, Educação e Educação Inclusiva, a fim de mapear as contribuições teóricas e desafios metodológicos para a atualização sistemática de repertórios que promovam novas construções epistemológicas para a Educação Inclusiva. MÉTODO: O estudo baseou-se em uma busca de artigos científicos publicados no SciELO entre os anos de 2009 e 2019, a partir dos descritivos de busca "neuropsicologia e educação" e "neuropsicologia e educação inclusiva". A partir da seleção de 21 artigos, empreendeu-se uma análise textual que possibilitou a categorizações temáticas, a partir dos seguintes descritores de análise: "Favorecendo novos/outros contextos sociais", "Uso x abuso de medicamentos", "Empenho/Desempenho Cognitivo: do que estamos falando"; "Funções executivas: avanços necessários"; "Testagem x estereotipagem social". RESULTADOS: Como resultado, evidenciamos que, quantitativamente, a maioria dos estudos ocupam os domínios das ciências da saúde, provenientes de pesquisas experimentais de enfoque cognitivista, enquanto uma minoria ocupa periódicos de ciências sociais/humanas e educação. Não obstante aos limites teóricos e conceituais que reproduzem uma leitura fragmentada de desenvolvimento humano, reforçando um enfoque diagnóstico que acarreta abuso de medicamentos como técnica de ajustamento, há proposições pautadas na Psicologia Histórico-Cultural que, qualitativamente, constituem-se como possibilidade de alinhamento da Neuropsicologia a ações educacionais inclusivas. CONCLUSÃO: Concluímos que, para o desafio de anúncio na democratização de oportunidades educacionais, é preciso avançar no desvelamento das condições materiais que sustentam os pilares de uma sociedade capitalista periférica, bem como os impactos psicossociais implicados nesta, na determinação de nossas funções psíquicas superiores.

INTRODUCTION: This article aims to analyze the thematic link between Neuropsychology, Education and Inclusive Education, in order to map the theoretical contributions and methodological challenges for the systematic updating of repertoires that promote new constructions for inclusive education. METHODS: The study was based on a search for scientific articles published in SciELO between the years 2009 and 2019 based on the search descriptions "neuropsychology and education" and "neuropsychology and inclusive education". Based on the selection of 21 articles, a textual analysis was carried out that enabled the thematic categorization, based on the following analysis descriptors: "Favoring new / other social contexts", "Use x abuse of medicines", "Commitment / Cognitive Performance: what are we talking about"; "Executive functions: necessary advances"; "Testing vs. social stereotyping". RESULTS: As a result, we show that quantitatively, most studies occupy the domains of health sciences, derived from experimental research with a cognitive focus, while a minority occupies journals in the social / human sciences and education. Despite the theoretical and conceptual limits that reproduce a fragmented reading of human development, reinforcing a diagnostic focus that leads to drug abuse as an attempt to adjust, there are propositions based on Historical-Cultural Psychology that, qualitatively, constitute a possibility of aligning the Neuropsychology to inclusive educational actions. CONCLUSION: We conclude that for the advertising challenge in the democratization of educational opportunities, it is necessary to advance in the unveiling of the perverse material conditions that support the pillars of a peripheral capitalist society, as well as the psychosocial impacts implied in this, in the determination of our higher psychic functions.

Survival, development and reproduction of Harmonia axyridis (Pallas, 1773) (Coleoptera: Coccinellidae) under effects of insecticides used in tomato plants.

Agriculture depends on the correct use of different methods for pest control, however the cultivation methods adopted for Solanaceae demands numerous pesticide applications. These products can be either harmful or selective to beneficial organisms, such as Coccinellidae predators. The aim of this study was to assess the physiological selectivity of insecticides, registered for tomato pest control, on Harmonia axyridis (Pallas, 1773) (Coleoptera: Coccinellidae). The maximum recommended field dosage of the insecticides spinetoram, spiromesifen, methoxyfenozide, metaflumizone, tebufenozide and methomyl was used. They were sprayed on eggs, third instar larvae, pupae and adults of H. axyridis using Potter´s tower; control treatment consisted of water. Methoxyfenozide didn't reduce any biological feature of the predator. Spiromesifen only reduced survival over time when sprayed on third instar larvae. Spinetoram shortened larval period and reduced survival of adults. When sprayed on larvae, metaflumizone reduced larval survival. Tebufenozide reduced egg viability, pupal period and larval survival over time. It is concluded that methoxyfenozide, spiromesifen and spinetoram are the least toxic to H. axyridis, and their usage should be prioritized in integrated pest management (IPM) programs; both tebufenozide and metaflumizone are harmful to some important biological features of the predator. Methomyl is highly toxic in controlled conditions, so its toxicity to H. axyridis should be confirmed in field and greenhouse tests, we also suggest further studies with the other compounds in different concentration and route of exposure.

Evaluation of Mycotoxin Production and Phytopathogenicity of the Entomopathogenic Fungi Fusarium caatingaense and F. pernambucanum from Brazil.

Fusarium incarnatum-equiseti species complex (FIESC) is considered as one of the richest insecticolous species. Fusarium species synthesize toxic secondary metabolites that are not fully understood. Mycotoxin production and pathogenicity on germinating seeds, seedlings, and leaves must be carefully studied for the use of Fusarium species in the biological control of insect pests. In this study, we evaluated the mycotoxin production and phytopathogenic potential of entomopathogenic strains of Fusarium sulawesiensis (1), F. pernambucanum (3), and F. caatingaense (23). The phytopathogenicity tests of F. caatingaense (URM 6776, URM 6777, URM 6778, URM 6779, and URM 6782) were performed during the development of bean (Phaseolus vulgaris, Vigna unguiculata, and Phaseolus lunatus), and corn (Zea mays) seedlings, using four treatments (soil infestation with the inoculum, spraying on leaves, root dip, and negative control). The mycotoxins, monoacetyl-deoxynivalenols (AcDON), deoxynivalenol (DON), beauvericin (BEA), fusarenone-X (FUS), T-2 toxin (T2), diacetoxyscirpenol (DAS), and zearalenone (ZEA), were detected in the study; BEA (detected in 25 strains) and FUS (detected in 21 strains) were found to be predominant. None of the strains showed any ability to cause disease or virulence in beans and corn. The FIESC strains showed a highly variable production of mycotoxins without the potential to be used as phytopathogenic agents for the cultures tested.

Evaluation of cytotoxic, immunomodulatory effects, antimicrobial activities and phytochemical constituents from various extracts of Passiflora edulis F. flavicarpa (Passifloraceae).

The leaves of P. edulis were subjected to physicochemical analysis, such as ion content, extractives, and structural molecules. The hexanic, ethanolic and ethyl acetate extracts were submitted to phytochemical analyzes by GC-MS, HPLC-MS, and spectrophotometry. In addition, antioxidant (DPPH, ABTS and TAA methods) potential, antimicrobial (MIC method) action, cytotoxicity and immunostimulant activity (flow cytometry analysis) were performed. The extracts showed a moderate antioxidant capacity and revealed the presence of several metabolites, mainly phenols, such as caffeic acid, p-coumaric acid and luteolin. The ethyl acetate and ethanolic extracts showed antifungal activity. In addition, the extracts did not affect splenocytes viability at 12.5 µg/mL and promoted the production of IL-6, IL-10, IL-17 and TNF-α cytokines. P. edulis extracts showed antifungal and antioxidant activity and were able to induce immunostimulatory action in splenocyte cultures in vitro.

Organizing the flow of physiotherapy services: agreement between clinical perception and a referral protocol / Organização do fluxo de serviços fisioterapêuticos: concordância entre percepção clínica e protocolo

Abstract Introduction: Protocols to organize the flow of treatment between primary and secondary healthcare levels help physiotherapists working in Primary Health Care (PHC) determine which cases will remain at the primary level and which should be referred to the secondary level for specialized treatment. Objective: Assess the agreement between the clinical perception of physiotherapists and the protocol in organizing the flow of physiotherapy patients. Methods: This is a methodological cross-sectional study based on the analysis of secondary data, recorded on a service spreadsheet, with respect to the clinical perception of 4 physiotherapists working in PHC and the protocol they apply to determine the urgency for referral to the secondary care level, considering physiotherapy specialties. Assessment was expressed as the percentage agreement, magnitude and significance according to the Kappa test, with > 0.80 considered perfect agreement. Analyses were conducted using SPSS 21.0 statistical software at a significance level of α = 0.05. Results: In 619 of 715 referrals, the perception of physiotherapists corroborated with the protocol in terms of patient referral to the secondary service. The percentage agreement for urgency classification in the total sample was 71% and the Weighted Kappa index 0.3710 (CI95% 0.3029-0.4391). The instrument exhibited high agreement in the areas of musculoskeletal physiotherapy (94.7%) and gerontology (98.2%), and low in urogynecology (27.6%). Conclusion: The protocol showed a high percentage of agreement and may be an important instrument in organizing the flow of physiotherapy services and could be enhanced for use in other specialties.

Resumo Introdução: Protocolos para organização do fluxo dos atendimentos entre os níveis primário e secundário de atenção ajudam o fisioterapeuta atuante na Atenção Primária à Saúde (APS) a determinar quais casos serão mantidos no primeiro nível e quais devem ser encaminhados para o nível secundário, onde receberão atendimento especializado. Objetivo: Avaliar a concor-dância entre percepção clínica dos fisioterapeutas e protocolo na organização do fluxo de usuários de serviço de fisioterapia. Métodos: Estudo transversal metodológico baseado em análise de dados secundários, registrados em planilha do serviço, referentes à percepção clínica de quatro fisioterapeutas atuantes na APS e protocolo aplicado por eles para determinação do encaminhamento e caráter de urgência para o nível secundário da rede de atenção, considerando as especialidades da fisioterapia. A avaliação expressou-se pelo percentual de concordância, magnitude e significância pelo teste Kappa, considerando > 0,80 concordância perfeita. As análises foram realizadas no pacote estatístico SPPS 21.0, nível de significância α = 0,05. Resultados: Considerando 715 encaminhamentos, em 619 a percepção dos fisioterapeutas corroborou com a determinação do protocolo em relação ao encaminhamento do usuário para o serviço secundário. O percentual de concordância relativo à classificação urgência, na amostra total, foi de 71% e o índice de Kappa Ponderado foi 0,3710 (IC95% 0,3029-0,4391). O instrumento apresentou maior concordância nas áreas de muscoloesquelética (94,7%) e gerontologia (98,2%), e menor concordância na uroginecologia (27,6%). Conclusão: O protocolo apresentou importante percentual de concordância, podendo ser instrumento importante na organização de fluxos de serviços de fisioterapia e ser aprimorado para o uso em várias especialidades.

CD8+ T cells are present at low levels in the white matter with physiological and pathological aging.

The presence and functional role of T cell infiltration in human brain parenchyma with normal aging and neurodegeneration is still under intense debate. Recently, CD8+ cells have been shown to infiltrate the subventricular zone in humans and mice with a deleterious effect on neural stem cells. However, to which extent T cell infiltration in humans also occurs in other regions such as cortical areas and, especially, white matter (WM) has not yet been addressed. In this work, we report a low-grade infiltration of T cells (CD3+, CD4+ and CD8+) in the WM of aged individuals that is also observed at similar levels in patients with neurodegenerative disorders (Alzheimer´s disease). In particular, CD3+ and CD8+ cells were increased in perivascular and parenchymal WM and cortical regions (enthorinal cortex). These results reveal that T cell infiltration in brain parenchyma occurs with physiological and pathological aging in several regions, but it seems to be lower than in the subventricular zone neurogenic niche.

CD8+ T cells are increased in the subventricular zone with physiological and pathological aging.

Age-related cognitive decline and neurodegenerative diseases are associated with less functional neurogenic niches. It has been recently shown that aged subventricular zone (SVZ) suffers an infiltration of T cells, which affects neural stem cell activity in mice. Whether this occurs in human neurogenic niches or to which extent T-cell infiltration is also taking place in neurodegenerative diseases remains unknown. In this work, we studied the presence of T cells in both human neurogenic niches in young and old individuals. There was a significant increase in the number of CD3+ and CD8+ T cells in the SVZ of elderly individuals, which was not detected in the dentate gyrus. Moreover, we also found CD3+ and CD8+ T cells in the SVZ of individuals with neurodegenerative diseases. However, T-cell count was similar when compared non-neuropathological elderly with disease diagnosed patients. Our study reveals the infiltration of T cells in old human brains, particularly in the SVZ under non-pathological conditions and also in neurodegenerative contexts.

Susceptibility of Chrysoperla externa (Hagen, 1861) (Neuroptera: Crysopidae) to insecticides used in coffee crops.

The coffee crop hosts pests such as mites, mealybugs, and aphids which serve as food for the predator Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae). The preservation of this chrysopid in coffee agroecosystem is very important to achieve sustainability of this agricultural sector, and can be obtained by applying low toxicity insecticides. The present study aimed to evaluate the susceptibility of C. externa to azadiracthin, chlorpyrifos, ethiprole and teflubenzuron. Predator eggs, third instar larvae, pupae and adults were exposed to insecticides by Potter tower spraying. When evaluating exposure of C. externa eggs we observed that chlorpyrifos, ethiprole and teflubenzuron reduced larvae hatching, while azadiracthin prolonged first instar duration. Meanwhile, the exposure of third instar larvae to chlorpyrifos and ethiprole caused mortality of all insects after 72 h, while azadiracthin prolonged the larval development time; we also observed that no compound allowed the formation of adults. After pupae were exposed to chlorpyrifos and teflubenzuron, it was observed a reduction on the emergence of adults, while the longevity of adults from these pupae and the evaluated reproductive parameters were reduced by all insecticides. For the bioassay with adults, chlorpyrifos, ethiprole and teflubenzuron reduced the longevity of insects, while the reproductive parameters evaluated were negatively affected after exposure to azadiracthin and teflubenzuron. It was concluded that all insecticides negatively affected at least one biological characteristic of the predator in at least one of the insect's developmental stages, requiring further research in semi-field and field conditions to prove its toxicity.

Bacillus thuringiensis endotoxin production: a systematic review of the past 10 years.

Bacillus thuringiensis (Bt) is one of the most promising biological control agents used commercially. Its products can contribute to reducing ecological and environmental problems associated with the use of chemical pesticides. Among the limiting factors of using Bt as bioinsecticide are the costs and ensuring its biological activity, which may vary according to the strain and culture conditions. This systematic review aimed to collect state-of-the-art information on the production of Bt endotoxins and to score the methodological feasibility of the data obtained, thus highlighting possible incoherencies. In order to consolidate recent findings and guide future studies, a total of 47 original articles from the last 10 years was analysed, with special attention being given to corroborating data, identifying inconsistencies and suggesting future adjustments so as to increase data reliability. With a maximum score of 8 points, three production parameters were classified on the following scale: preferable (score: 2), adequate (score: 1) and inadequate (score: 0), and another two parameter were classified as adequate (score: 1) or inadequate (score: 0). No article scored more than 6 out of the maximum of 8, thus reflecting the need for more detailed studies regarding Bt endotoxin production. The lack of standardization of methods and units of measurement also have made a comparison of results and an overall analysis difficult. Standards are suggested in the present study. The inclusion of bioassays and quantifying toxin via alkaline dilution are strongly recommended for studies of this nature, along with LC50 expressed in mg/L. Sixteen articles (34%) did not use either of these suggested methods, which indicates the need for further supporting studies. These findings reinforce the need for robust studies in this area, which could include the development of more affordable and effective bioinsecticides, thus increasing their competitiveness against insecticides derived from unsustainable sources.

Effects of Cefazolin and Meropenem in Eradication Biofilms of Clinical and Environmental Isolates of Proteus mirabilis.

Proteus mirabilis is an opportunistic Gram-negative bacterium belonging to the family Enterobacteriaceae and is known for its ability to cause urinary tract infections. The aim of this study was to determine the value of the minimum concentration of cefazolin and meropenem on biofilm eradication, as well as the resistance profiles and genetic diversity of clinical and environmental isolates of P. mirabilis. We compared the isolates collected from a hospital environment and from an urban stream impacted in Recife-Pernambuco, Brazil. Biochemical tests were performed to determine the profiles of susceptibility, hydrophobicity, biofilm formation and eradication. The genetic diversity was verified using the ERIC-PCR method. The results revealed that two clinical isolates (ICP4 and ICP5) were multi-drug resistant, whereas the environmental isolates showed resistance only to tetracycline, except for CP525S, which was resistant also to ampicillin. Of the isolates investigated, three were moderately hydrophobic, while the remaining were hydrophilic. Genetic diversity analysis verified the presence of clones indicating that the stream is harboring and disseminating bacteria of hospital origin. All isolates formed a biofilm, however, high concentrations of cefazolin and meropenem were required to eradicate the already formed biofilm. Our study analyzed the survival strategies of these bacteria in the environments investigated and corresponds to first report the use of these antibiotics to eliminate P. mirabilis biofilms.