A critical review of adverse effects to the kidney: mechanisms, data sources, and in silico tools to assist prediction.

INTRODUCTION: The kidney is a major target for toxicity elicited by pharmaceuticals and environmental pollutants. Standard testing which often does not investigate underlying mechanisms has proven not to be an adequate hazard assessment approach. As such, there is an opportunity for the application of computational approaches that utilize multiscale data based on the Adverse Outcome Pathway (AOP) paradigm, coupled with an understanding of the chemistry underpinning the molecular initiating event (MIE) to provide a deep understanding of how structural fragments of molecules relate to specific mechanisms of nephrotoxicity. Aims covered: The aim of this investigation was to review the current scientific landscape related to computational methods, including mechanistic data, AOPs, publicly available knowledge bases and current in silico models, for the assessment of pharmaceuticals and other chemicals with regard to their potential to elicit nephrotoxicity. A list of over 250 nephrotoxicants enriched with, where possible, mechanistic and AOP-derived understanding was compiled. Expert opinion: Whilst little mechanistic evidence has been translated into AOPs, this review identified a number of data sources of in vitro, in vivo, and human data that may assist in the development of in silico models which in turn may shed light on the interrelationships between nephrotoxicity mechanisms.

Cross-talk between lung cancer and bones results in neutrophils that promote tumor progression.

Lung cancer is the leading cause of cancer mortality around the world. The lack of detailed understanding of the cellular and molecular mechanisms participating in the lung tumor progression restrains the development of efficient treatments. Recently, by using state-of-the-art technologies, including in vivo sophisticated Cre/loxP technologies in combination with lung tumor models, it was revealed that osteoblasts activate neutrophils that promote tumor growth in the lung. Strikingly, genetic ablation of osteoblasts abolished lung tumor progression via interruption of SiglecFhigh-expressing neutrophils supply to the tumor microenvironment. Interestingly, SiglecFhigh neutrophil signature was associated with worse lung adenocarcinoma patients outcome. This study identifies novel cellular targets for lung cancer treatment. Here, we summarize and evaluate recent advances in our understanding of lung tumor microenvironment.

In vitro evaluation of biomarkers of nephrotoxicity through gene expression using gentamicin.

Acute renal failure is one of the most frequent effects observed after taking medicine. Such situations have been tardily discovered, given that existing methods for assessing toxicity are not predictive. In this light, the present work evaluated the effects of gentamicin, a form of nephrotoxic drug, on HK-2 and HEK-293 cells. By using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and flow cytometry, both cells demonstrated that cytotoxicity occurs in a dose-dependent manner through the processes of apoptosis and cell necrosis. Gene expression analysis showed a relative increase of expression for genes related to cell processes and classic biomarkers, such as TP53, CASP3, CASP8, CASP9, ICAM-1, EXOC3, KIM-1, and CST3. A decrease in expression for genes BCL2L1 and EGF was observed. This study, therefore, indicates that, when the methods are used together, gene expression analysis is able to evaluate the nephrotoxic potential of a substance.

Biodistribution, pharmacokinetics and toxicity of a Vasconcellea cundinamarcensis proteinase fraction with pharmacological activity

Abstract Prior studies demonstrate that a proteinase fraction from Vasconcellea cundinamarcensis V.M. Badillo, Caricaceae, exhibits wound healing activity in gastric and cutaneous models and antitumoral/antimetastatic effects. Here, we present the toxicity, pharmacokinetics and biodistribution data for this proteinase fraction following a single dose into Swiss mice by i.v., s.c. or p.o. routes. The i.v. and s.c. toxicity assays demonstrate that proteinase fraction at ≤20 mg/kg is non-lethal after single injection, while parental administration (p.o.) of ≤300 mg/kg does not cause death. Based on p.o. acute toxicity dose using Organisation for Economic Cooperation and Development protocols, proteinase fraction ranks as Class IV “harmful” substance. Proteinase fraction shows high uptake determined as Kp (distribution tissue/blood) in organs linked to metabolism and excretion. Also, high bioavailability (≈100%) was observed by s.c. administration. The blood contents following i.v. dose fits into a pharmacokinetic bi-compartmental model, consisting of high removal constants – kel 0.22 h−1 and kd 2.32 h−1and a half-life – t½ = 3.13 h. The Ames test of proteinase fraction (0.01–1%) demonstrates absence of mutagenic activity. Likewise, genotoxic evaluation of proteinase fraction (5 or 10 mg/kg, i.p.) shows no influence in micronuclei frequency. In conclusion, the acute doses for proteinase fraction lack mutagenic and genotoxic activity, clearing the way for clinical assays.

Toxicological evaluation of the João da Costa e Associações® on Beagle dogs

A toxicological study was performed in Beagle dogs treated for 180 days with the product João da Costa e Associações. Were used six males and six females distributed in control and treated groups (n=3). We used a dose of 566 mg/kg of the product according to preclinical study in rodents. The animals were weighed and evaluated by clinical and laboratory aspects. The product did not cause mortality or alter the normal behavior of animals, but interfered with the weight gain on males in the middle phase of the treatment. The group treated had a lower incidence of clinical abnormalities compared to control, checked by veterinary consultations. Laboratory data showed elevated blood glucose levels perhaps due to the high amount of sucrose present in the product; about the histopathological data no significant change was found. We conclude that the product Joao da Costa and Associações, at the dose tested, has low toxicity in Beagle dogs treated chronically.

A comparison of BGM and LLC-PK1 cells for the evaluation of nephrotoxicity.

Nephrotoxicity is one of the most frequent effects observed after the use of medicine. Such situations have been tardily discovered because of existing methods to determine toxicity. The validation of sensitive, alternative methods for the early identification of toxic effects is as important as restrictions on the use of animals. In this light, the present study evaluated the effects of gentamicin on BGM and LLC-PK1 cells, using MTT and Neutral Red (NR). Although the LLC-PK1 cell line is used for toxicological studies, the BGM cell line is relatively new for this purpose. MTT (BGM: EC(50) = 6.29 mM; LLC-PK1: EC(50) = 8.01 mM) was found to be more sensitive than NR (EC(50) was greater than 10 mM for both cells). By using MTT, both cells demonstrated the involvement of mitochondria in a manner that was dose dependent, with an apoptotic process occurring at the concentrations of 1 and 3 mM and necrosis at concentrations above 4 mM. It could, therefore, be concluded that 1) BGM appears to be useful in the study of the mechanism of nephrotoxicity caused by gentamicin and 2) because of its sensitivity to MTT, in addition to its ease of manipulation, it is believed that the BGM cell line can also be used as an alternative method to evaluate nephrotoxicity.

Evaluation of the immunotoxicological effects of Dimorphandra mollis Benth., Fabaceae, in rats / Avaliação dos efetos imunotoxicológicos de Dimorphandra mollis Benth., Fabaceae, em ratos

Dimorphandra mollis Benth., Fabaceae, also known as "faveira" or "fava-d'anta", is a plant common to the central woodsy meadow region of Brazil. It is well known for its antioxidant, antiplatelet and, principally, vasoprotective properties. Its principal component is rutin. The objective of this study is the evaluation of the safety of the use of the dried D. mollis extract in rodents. The rutin content of the standardized extract was 76.0±3 percent. With respect to the biochemical and hematological parameters evaluated, no alterations in the groups of rats that received 1000 and 2000 mg/kg doses of D. mollis were observed, but an increase in eosinophiles occurred. Hyperactivity of the white splenic pulp was detected in the group that received the 2000 mg/kg dose of D. mollis. In the evaluation of the lymphproliferative response with 1000 and 2000 mg/kg, no alterations were observed, and a decrease in IgG was only observed in the studies with a 2000 mg/kg dose. The results obtained with rodents suggest that no toxicity exists with the administration of dried D. mollis extract in a 1000 mg/kg dose.

A Dimorphandra mollis Benth., Fabaceae, conhecida como faveira ou fava-d'anta, é uma planta comum do cerrado central do Brasil, muito utilizada por suas propriedades antioxidante, antiplaquetária e, principalmente, como vasoprotetora. Seu principal marcador é a rutina. Este estudo teve como objetivo avaliar a segurança da utilização do extrato seco de D. mollis em roedores. O extrato foi extraído, padronizado e quantificado apresentando teor de 76,0±3 por cento de rutina. Nos parâmetros bioquímicos e hematológicos avaliados, não se observou alterações nos grupos de machos e fêmeas que receberam a dose de 1000 e 2000 mg/kg de D. mollis, mas observou-se um aumento de eosinófilos. Nos estudos histopatológicos detectou-se hiperreatividade da polpa branca esplênica, no grupo que recebeu a dose de 2000 mg/kg de D. mollis. Na avaliação da resposta linfoproliferativa, com 1000 e 2000 mg/kg não foram observadas alterações, e somente nos estudos com a dose de 2000 mg/kg se observou diminuição de IgG. Os resultados obtidos, utilizando roedores, sugerem que nenhuma toxicidade existe na administração de extrato seco de D. mollis na dose de 1000 mg/kg.

Influence of the preparation method on the low efficacy of multi-herb commercial products: the example of João da Costa e Associações / Influência do método de preparação na baixa eficácia de produtos fitoterápicos: o exemplo do João da Costa e Associações

Native medicinal plants have been used for decades by Brazilian pharmaceutical companies to create commercial products. In this study, we have investigated the herb-combined product João da Costa e Associações® (JCA) commercialized for thirty years to treat dysmenorrhoea. JCA is prepared by decoction of Himatanthus lancifolius (Muell. Arg.) Woodson (Apocynaceae), Chondodendron platyphyllum Miers (Menispermaceae), Gossypium herbaceum L. (Malvaceae), Rosmarinus officinalis L. (Lamiaceae) and Echites peltata (Apocynaceae), followed by addition of sugar. The efficacy of JCA was verified by antinociceptive studies. The chemical composition was determined by fingerprint analysis in HPLC/ DAD. A weak inhibition of the second phase of the nociceptive effect induced by formalin indicated an activity similar to those steroids and not-steroids anti-inflammatories. Despite being prepared by decoction of five plants, the fingerprint analysis showed only two peaks. None of them corresponds to the chemical compounds observed in ethanol extracts prepared with the same plant material. We argue that the methods of preparation of the formulas should be considered in studies of multi-herbs products, since they can be the responsible for inefficacy or low activity of such products.

Plantas medicinais nativas do Brasil foram usadas por décadas pelas indústrias farmacêuticas nacionais para criar seus produtos. Neste estudo, foi investigado o produto João da Costa e Associações® (JCA) comercializado por mais de trinta anos para o tratamento de dismenorréia e outros problemas relacionados à saúde da mulher. JCA é preparado pela decocção de Himatanthus lancifolius (Muell. Arg.) Woodson (Apocynaceae), Chondodendron platyphyllum Miers (Menispermaceae), Gossypium herbaceum L. (Malvaceae), Rosmarinus officinalis L. (Lamiaceae) e Echites peltata (Apocynaceae), seguido de adição de açúcar. A eficácia de JCA foi verificada por meio da avaliação da atividade antinociceptiva. Já a composição química foi determinada por analises em HPLC/ DAD. Uma fraca inibição da segunda fase da nocicepção foi observada no teste da formalina, indicando uma ação semelhante aos antiinflamatórios esteroidais e não esteroidais. Apesar de ser preparado pela decocção de cinco plantas, a análise no HPLC apresentou somente dois picos, e nenhum deles correspondeu aos componentes observados nos extratos etanólicos preparados com as mesmas plantas. Os resultados sugerem que o método de preparação de JCA promove a perda dos componentes químicos das plantas e interfere consideravelmente na eficácia do produto.

Acute and chronic toxicological studies of the Brazilian phytopharmaceutical product Ierobina / Estudos de toxicidade aguda e crônica do produto fitoterápico brasileiro Ierobina

Ierobina® is a Brazilian phytopharmaceutical product employed for the treatment of dyspepsia (280 mg/kg/day). Despite its widespread use in the country for over 75 years, only recently its therapeutic efficacy has been attested in animals; however, no toxicological investigations have been carried out for the product to date. In this paper we evaluated the acute toxicity of Ierobina® administrated by gavage in mice (single doses of 2100 mg/kg, 6300 mg/kg and 12600 mg/kg), along with its chronic effects in rats, after product administration per os daily, at the doses of 2800 mg/kg and 5600 mg/kg, for 180 days. The product had low acute toxicity; all observed alterations were reversible and no animal died during the experiments. In chronic toxicological studies, Ierobina® administration for 180 days did not cause any changes in hematological and biochemical parameters, with the exception of decreasing the levels of alanine transaminase, aspartate transaminase and creatinine. However, histological evaluation of kidney, liver and other selected organs showed normal architecture, suggesting no morphological disturbances. Hence, considering the obtained results and the fact that Ierobina® has been commercialized for decades in Brazil, without any notified case of toxicity, it seems that the product is safe for human use.

A Ierobina® é um produto utilizado popularmente, no Brasil, para tratamento de dispepsia, na dose de 280 mg/kg/dia. Apesar de seu largo uso nos últimos 75 anos, recentemente foi comprovada sua eficácia em animais; porém, nenhuma avaliação de seu perfil toxicológico foi realizada. O objetivo do presente estudo foi avaliar a toxicidade aguda (doses únicas de 2100 mg/kg, 6300 mg/kg ou 12600 mg/kg), em camundongos, e crônica (doses de 2800 mg/kg ou 5600 mg/kg, por 180 dias), em ratos, após administração per os de Ierobina®. No teste de toxicidade aguda, as doses administradas não produziram nenhuma mortalidade e os sinais observados foram todos reversíveis. No teste de toxicidade crônica, não foram verificadas diferenças significativas nas análises hematológicas, macroscópicas e microscópicas. Nos exames de bioquímica sérica, diferença significativa foi observada somente na avaliação da alanina transaminase, aspartato transaminase e creatinina, porém, sem importância clínica. Assim, considerando os resultados obtidos e o fato de ser a Ierobina® um produto comercializado há décadas, sem qualquer notificação de casos de toxicidade, podemos concluir que o produto parece ser de segurança adequada para uso humano.

Anti-inflammatory activity and gastric lesions induced by zinc-tenoxicam.

Oral administration of tenoxicam or zinc-tenoxicam complex inhibited to a similar extent carrageenin-induced paw oedema and granulomatous tissue formation in rats as well as the acetic acid induced writhing response in mice. Gastric lesions induced by oral administration of zinc-tenoxicam were reduced in number and severity when compared with those induced by tenoxicam or the co-administration of tenoxicam and zinc acetate. However, after intraperitoneal administration, both zinc-tenoxicam and tenoxicam plus zinc acetate induced a reduced number of gastric lesions as compared with tenoxicam.