XinJiaCongRongTuSiZiWan protects triptolide-induced rats from oxidative stress injury via mitophagy mediated PINK1/Parkin signaling pathway

Purpose: XinJiaCongRongTuSiZiWan (XJCRTSZW) is a traditional Chinese medicine compound for invigorating the kidney, nourishing blood, and promoting blood circulation. This study aimed to explore the effect of XJCRTSZW on triptolide (TP)-induced oxidative stress injury. Methods: Adult female Sprague-Dawley rats and human ovarian granulosa cell lines were treated with TP and XJCRTSZW. Hematoxylin and eosin staining, enzyme-linked immunosorbent assay, flow cytometry, CCK-8, JC-1 staining, transmission electron microscopy, reverse transcription-quantitative polymerase chain reaction, and Western blotting were performed in this study. Results: XJCRTSZW treatment observably ameliorated the TP-induced pathological symptoms. Furthermore, XJCRTSZW treatment observably enhanced the TP-induced reduction of estradiol, anti-Mullerian hormone, progesterone, superoxide dismutase, ATP content, mitochondrial membrane potential, p62, and Hsp60 mRNA, and protein levels in vivo and in vitro (p < 0.05). However, TP-induced elevation of follicle stimulating hormone and luteinizing hormone concentrations, malondialdehyde levels, reactive oxygen species levels, apoptosis rate, mitophagy, and the mRNA and protein expressions of LC3-II/LC3-I, PTEN-induced kinase 1 (PINK1), and Parkin were decreased (p < 0.05). In addition, XJCRTSZW treatment markedly increased cell viability in vitro (p < 0.05). Conclusions: XJCRTSZW protects TP-induced rats from oxidative stress injury via the mitophagy-mediated PINK1/Parkin pathway.

5HT2A modulation attenuates pancreatic cancer induced pain mouse model by inhibiting HDAC

Purpose Patients have been severely suffered from cancer associated pain, and pancreatic cancer is the most severe form of cancer associated with pain. There are very few options available to manage it. The present report evaluated the effect of 5HT2A on pancreatic cancer associated pain. Methods Pancreatic cancer was induced by injecting SW 1,990 cells (~3×106 in a 20 µL suspension) into the pancreas and formed a 2­3-mm vesicle using an inoculator fitted with a 26-gauge needle in BALB/c-nu mice. Survival rate and body weight of the mice were observed. Pain behaviour testing was performed at the end of each week (third and fourth week) after surgery. Inflammatory mediators and HDAC 2 proteins were determined in the spinal tissue using quantitative real-time polymerase chain reaction. Results There was improvement in the survival rate and body weight in 5HT2A antagonist treated group than pancreatic cancer group of mice. Moreover, 5HT2A antagonist ameliorated the alteration in pain behaviour of pancreatic cancer mice. mRNA expression of HDAC2 and level of inflammatory cytokines were reduced in the spinal tissue of 5HT 2A antagonist treated group than pancreatic cancer group of mice. Conclusions Data revealed that 5HT2A antagonist ameliorates pain associated with pancreatic cancer mice by HDAC inhibition and inflammatory cytokines. The result of investigation supports that modulation of 5HT2A receptor could be used clinically to protects neuropathic pain in pancreatic cancer.

Establishment and application of a new 4/6 infarct nephrectomy rat model for moderate chronic kidney disease

Purpose: To develop a new 4/6 infarct nephrectomy (INx) model rat mimicking moderate chronic kidney disease (CKD) and to evaluate its application. Methods: We modified the conventional 5/6 INx rat model to create the 4/6 INx model by ligating the renal artery branch to induce infarction of one-third of the left kidney after right kidney removal and compared biochemically and histologically both models. To demonstrate the application of the 4/6 INx model, the effects of a supplementary compound containing calcium carbonate, chitosan, palm shell activated charcoal etc., that is effective for both CKD and its complications, were compared between both models. Results: Impairment of renal function in the 4/6 INx group was significantly more moderate than in the 5/6 INx group (P < 0.05). The 4/6 INx group showed less histological damage in kidney than in the 5/6 INx group. The supplementary compound did not improve CKD in the 5/6 INx group, but ameliorated elevation of blood urea nitrogen in the 4/6 INx group. Conclusions: We developed the 4/6 INx model, which is more moderate than the conventional 5/6 INx model. This model could potentially demonstrate the effectiveness of drugs and supplements intended to prevent CKD and its progression.

Antiosteoporosis effect of bryodulcosigenin on ovariectomy-induced osteoporosis in experimental rats

Purpose: Osteoporosis is a bone disease which commonly occurred in postmenopausal women. Almost 10 percent of world population and approximately 30% of women (postmenopausal) suffer from this disease. Alternative medicine has great success in the treatment of osteoporosis disease. Bryodulcosigenin, a potent phytoconstituent, already displayed the anti-inflammatory and antioxidant effect. In this study, we made effort to analyze the antiosteoporosis effect of bryodulcosigenin against ovariectomy (OVX) induced osteoporosis in rats. Methods: Swiss albino Wistar rats were grouped into fIve groups and given an oral dose of bryodulcosigenin (10, 20 and 30 mg/kg) for eight weeks. Body weight, uterus, bone mineral density, cytokines, hormones parameters, transforming growth factor (TGF)-ß, insulin-like growth factor (IGF), osteoprotegerin (OPG), receptor activator of nuclear factor kappa-Β ligand (RANKL), and its ratio were estimated. Results: Bryodulcosigenin significantly (p < 0.001) suppressed the body weight and enhanced the uterine weight and significantly (p < 0.001) increased the bone mineral density in whole femur, caput femoris, distal femur and proximal femur. Bryodulcosigenin significantly (P < 0.001) altered the level of biochemical parameters at dose dependent manner, significantly (P < 0.001) improved the level of estrogen and suppressed the level of follicle stimulating hormone and luteinizing hormone. Bryodulcosigenin significantly (P < 0.001) improved the level of OPG and suppressed the level of RANKL. Conclusions: Bryodulcosigenin reduced the cytokines level and suppressed the TGF-ß and IGF. We concluded that bryodulcosigenin is an antiosteoporosis medication based on the findings.

Protective effects of hesperidin in gastric damage caused by experimental ischemia-reperfusion injury model in rats

Purpose: This study evaluated the protective effect of hesperidin on injury induced by gastric ischemia-reperfusion. Methods: Fifty male Sprague Dawley rats (250­300 g) were divided into five groups: control (C), sham (S), ischemia (I), ischemia-reperfusion (I/R) and hesperidin + ischemia-reperfusion (Hes + I/R). Hesperidin was injected intraperitoneally at the dose of 100 mg/kg one hour before the experimental stomach ischemia-reperfusion. Celiac artery was ligated. After 45 minutes ischemia and 60 minutes reperfusion period, blood samples were obtained under anesthesia. Then, animals were sacrificed, stomach tissues were excised for biochemical, and histopathological analyses were performed. Malondialdehyde levels and superoxide dismutase, glutathione peroxidase activities and total antioxidant status (TAS), total oxidant status (TOS), protein, total thiol parameters were measured in plasma, and tissue homogenate samples. H + E, periodic acid­Schiff, hypoxia inducible factor, terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end-labeling (TUNEL), and proliferating cell nuclear antigen (PCNA) for cell proliferation as immunohistochemical parameters were determined. Results: Upon biochemical and histopathological assessment, hesperidin decreased stomach tissue changes in comparison with IR group. Ischemia-reperfusion injury led to a considerably increase in malondialdehyde, protein, and TOS levels (p < 0.001) in stomach tissue. Hesperidin treatment significantly decreased malondialdehyde, protein, and TOS levels (p < 0.001). Hesperidin increased superoxide dismutase, TAS, total thiol and glutathione peroxidase activities in comparison with IR group. Hesperidin reduced damage and also increased TUNEL and PCNA immunoreactivity in stomach tissue. Conclusions: Hesperidin was able to decrease I/R injury of the stomach tissue due to inhibition of lipid peroxidation and protein oxidation, duration of antioxidant, and free radical scavenger properties. Consequently, hesperidin can provide a beneficial therapeutic choice for preventing stomach tissue ischemia-reperfusion injury in clinical application.

A microscope in your pocket: can smartphones be used to perform microsurgery?

Purpose: To evaluate the use of the latest generation smartphone camera in performing arterial microanastomosis in rats. Methods: Ten Wistar rats were divided into 2 groups and underwent anastomosis of the right carotid artery with the aid of magnification from a microscope (group M) and a smartphone camera (group S), to compare patency in 72 hours, as well as to measure the weight of the animals, diameter of the carotid arteries and anastomosis time. Results: There was no statistical difference between the weight of the animals or the diameter of the carotid arteries. There was a statistical difference for the time spent on anastomoses, which was greater in group S, with higher rates of thrombosis (p < 0.05). Conclusions: Although our patency and anastomosis time results were statistically lower in the smartphone group, there was success in some cases. As the segment continues to progress, it is likely that the results will improve in line with the evolution of camera technology.

Temozolomide alleviates breast carcinoma via the inhibition of EGFR/ERK/ MMP-1 pathway with induction of apoptotic events

Purpose: To evaluate the chemotherapeutic activity of temozolomide counter to mammary carcinoma. Methods: In-vitro anticancer activity has been conducted on MCF7 cells, and mammary carcinoma has been induced in Wistar rats by introduction of 7, 12-Dimethylbenz(a)anthracene (DMBA), which was sustained for 24 weeks. Histopathology, immunohistochemistry, cell proliferation study and apoptosis assay via TUNEL method was conducted to evaluate an antineoplastic activity of temozolomide in rat breast tissue. Results: IC50 value of temozolomide in MCF7 cell has been obtained as 103 µM, which demonstrated an initiation of apoptosis. The temozolomide treatment facilitated cell cycle arrest in G2/M and S phase dose dependently. The treatment with temozolomide suggested decrease of the hyperplastic abrasions and renovation of the typical histological features of mammary tissue. Moreover, temozolomide therapy caused the downregulation of epidermal growth factor receptor, extracellular signal-regulated kinase, and metalloproteinase-1 expression and upstream of p53 and caspase-3 proliferation to indicate an initiation of apoptotic events. Conclusions: The occurrence of mammary carcinoma has been significantly decreased by activation of apoptotic pathway and abrogation of cellular propagation that allowable for developing a suitable mechanistic pathway of temozolomide in order to facilitate chemotherapeutic approach.

Eucommia granules activate Wnt/ß-catenin pathway, and improve oxidative stress, inflammation, and endothelial injury in preeclampsia rats

Purpose: Pre-eclampsia (PE) is a pregnancy-related complication. Eucommia is effective in the treatment of hypertensive disorders in pregnancy, but the specific effects and possible mechanisms of Eucommia granules (EG) in PE remain unknown. The aim of this study was to investigate the effects and possible mechanisms of EG in PE rats. Methods: Pregnant Sprague Dawley rats were divided into five groups (n = 6): the control group, the model group, the low-dose group, the medium-dose group, and the high-dose group of EG. The PE model was established by subcutaneous injection of levonitroarginine methyl ester. Saline was given to the blank and model groups, and the Eucommia granules were given by gavage to the remaining groups. Blood pressure and urinary protein were detected. The body length and weight of the pups and the weight of the placenta were recorded. Superoxide dismutase (SOD) activity and levels of malondialdehyde (MDA), placental growth factor (PIGF), and soluble vascular endothelial growth factor receptor-1 (sFIt-1) were measured in the placenta. Pathological changes were observed by hematoxylin-eosin staining. Wnt/ß-catenin pathway-related protein expression was detected using Western blot. Results: Compared with the model group, the PE rats treated with EG had lower blood pressure and urinary protein. The length and weight of the pups and placental weight were increased. Inflammation and necrosis in the placental tissue was improved. SOD level increased, MDA content and sFIt-1/PIGF ratio decreased, and Wnt/ß-catenin pathway-related protein expression level increased. Moreover, the results of EG on PE rats increased with higher doses of EG. Conclusions: EG may activate the Wnt/ß-catenin pathway and inhibit oxidative stress, inflammation, and vascular endothelial injury in PE rats, thereby improving the perinatal prognosis of preeclamptic rats. EG may inhibit oxidative stress, inflammation, and vascular endothelial injury through activation of the Wnt/ß-catenin pathway in preeclampsia rats, thereby improving perinatal outcomes in PE rats.

Ellagic acid protected the gingival tissue via fibroblast and epidermal growth factors in rats

Purpose: To investigate the effect of ellagic acid (EA) in gingival tissues injury in rats. Methods: Twenty rats were categorized into two groups. In burn group, an excisional wound area was created by removing a 4-mm diameter flap from the left molar region in the mucoperiosteal region of the gingiva. In burn + ellagic acid group, 1.2 mg/mL EA was administered as irrigation for one week. Animals was sacrificed under anesthesia at the end of experiment. Malondialdehyde (MDA), myeloperoxidase (MPO) and glutathione (GSH) level were measured. Hematoxylin and eosin, fibroblast growth factor (FGF) and epidermal growth factor (EGF) immunostainings were applied to tissues. Results: MDA, MPO, inflammation and leukocyte infiltration were high in burn group. Degeneration epithelium, edema and inflammatory cell infiltration in connective tissue areas, and dilatation and congestion in blood vessels were observed in burn group. In burn + EA group, the gingival epithelium improved, collagen fiber production increased and organized dermis were observed. After burn injury, FGF and EGF activity was increased in EA treated groups. Conclusions: We suggest that EA have the potential for better healing outcomes in oral wounds. EA seems to have promising therapeutic efficacy to enhance oral wound healing.

Effects of curcumin supplementation on abdominal surgical wound healing

Purpose: To evaluate the effects of curcumin supplementation on abdominal surgical wound healing in rats using clinical, histological, and hematological parameters. Methods: Forty Wistar rats were randomly divided into two groups: the curcumin group, and the control group. The curcumin group received, in addition to water and standard feed, curcumin via gavage at the dose of 200 mg/kg for seven days preceding and seven days following surgery. The control group received only water and standard feed. Both groups underwent median laparotomy and left colotomy. On the eighth postoperative day, the groups were euthanized, and the left colon was resected for histological analysis. Results: In the preoperative evaluation, there was a significant decrease in the mean C-reactive protein levels in the curcumin group (0.06) compared to the control group (0.112) (p = 0.0001). In the postoperative wound healing assessment, a significant decrease was observed in inflammatory infiltrate (p = 0.0006) and blood vessel count (p = 0.0002) in the curcumin group compared to the control group. Conclusions: Curcumin supplementation was able to significantly reduce inflammatory parameters in both pre-and post-operative phases of abdominal surgical wounds in rats.

Photobiomodulation effects on synovial morphology, iNOS gene, and protein expression in a model of acute inflammation

Purpose: To evaluate morphological aspects and inducible nitric oxide synthase (iNOS) gene and protein expression in a model of acute inflammation. Methods: Thirty-six female Wistar rats were assigned into three groups: control (saline, n = 12), sham (arthritis, n = 12), and PBM (arthritis and photobiomodulation, n = 12). Arthritis induction was performed with 200 µg of intra-articular Zymosan in sham and PBM animals. PBM was performed 24 h after induction with a laser device (λ = 808 nm, 25 mW of nominal power, fluence of 20 J/cm2, beam area of 0.02 mm2, time of 33 s, total energy of 0.825 J) with punctual and single dose application. Morphological analysis of joint structure (HE) and immunohistochemistry (anti-iNOS antibody) were performed on knee samples, and synovial tissue was submitted to RNA extraction, cDNA synthesis and gene expression analysis by quantitative polymerase chain reaction. Statistical analyses were performed with p < 0.05. Results: It was observed an increase in the thickness of the synovial lining epithelium and inflammatory infiltrate in sham compared to PBM. Gene expression analysis showed higher iNOS expression in PBM, and iNOS protein expression decreased in PBM compared to sham. Conclusions: Photobiomodulation decreased inflammation in PBM animals, upregulated iNOS gene expression, however down egulated protein expression compared to sham.

Avaliação do perfil de citocinas séricas e óxido nítrico na hanseníase experimental murina / Evaluation of the serum cytokines profile and nitric oxide in experimental murine leprosy

Introdução: a hanseníase é uma do-ença infecciosa crônica causada pelo Mycobacterium leprae (M. leprae), um para-sita intracelular obrigatório. Assim, a resis-tência do hospedeiro a esse patógeno depen-de da imunidade celular. O uso de modelos experimentais tem permitido o estudo da hanseníase do ponto de vista imunológico, microbiológico e terapêutico, entretanto, as diferenças na progressão da infecção entre os modelos mais empregados (camundongos imunocompetentes, BALB/c, e camundongos congenitamente atímicos, nude) são pouco estudadas. Objetivo: comparar a evolução da infecção pelo M. leprae em camundongos BALB/c e nude quanto à multi-plicação bacilar e avaliação do perfil inflamatório sistêmico pela quantificação sérica de citocinas e óxido nítrico (NO). Métodos: os camundongos foram inoculados com M. leprae nos coxins plantares e avaliados aos 3, 5 e 8 meses após a infecção. Resultados: camundongos nude apresentaram multiplicação bacilar progressiva nos coxins plantares. Em camundongos BALB/c, o número de bacilos foi maior aos 5 meses. Em relação à quantificação de citocinas, nos camundongos BALB/c houve aumento de IL-2 e IL-17A e diminuição de IL-6 e NO aos 8 meses de inoculação. Nos camundongos nude, verificou-se o aumento do TNF aos 8 meses de inoculação e manutenção dos níveis de NO. Conclusão: os resultados encontrados sugerem que em camundongos BALB/c ocorre a ativação de uma resposta imune capaz de controlar a multiplicação do M. leprae, em contrapartida em camundongos nude a infecção é progressiva a despeito de altos níveis de TNF. (AU)

Introduction: leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae), an obligate intracellular parasite. Thus, host resistance to this pathogen depends on cellular immunity. The use of experimental models has made it possible to study leprosy from an immunological, microbiological, and therapeutic point of view. However, the differences in the progression of the infection between the most used models (immunocompetent mice, BALB/c, and congenitally athymic mice, nude) have been little studied. Objective: to compare the evolution of M. leprae infection in BALB/c and nude mice in terms of bacillary multiplication and evaluation of the systemic inflammatory profile by quantifying serum cytokines and nitric oxide (NO). Methods: the mice were inoculated with M. leprae in the footpads and evaluated at 3, 5, and 8 months after infection. Results: nude mice showed progressive bacillary multiplication in the footpads. In BALB/c mice, the number of bacilli was higher at 5 months. In terms of cytokine quantification, BALB/c mice showed an increase in IL-2 and IL-17A and a decrease in IL-6 and NO at 8 months of inoculation. In the nude mice, there was an increase in TNF at 8 months of inoculation and maintenance of NO levels. Conclusion: the results suggest that BALB/c mice activate an immune response capable of controlling the multiplication of M. leprae, whereas in nude mice the infection is progressive despite high levels of TNF. (AU)

La hipertensión arterial en animales de laboratorio / High blood pressure in experimental animals

Las Ciencias Médicas y Biológicas requieren, prioritariamente, que la investigación y la experimentación sean desarrolladas sobre organismos completos (los modelos animales). Su utilización ha permitido desarrollar innumerables ensayos preclínicos para evaluar los mecanismos patógenos y terapéuticos de diversas enfermedades, así como el estudio de las causas, naturaleza y cura de múltiples desórdenes de la salud humana. En este trabajo se muestra una panorámica general de los biomodelos de hipertensión arterial donde se describen: conceptos, características, origen, importancia, utilidad y procedimientos experimentales durante su fase de inducción. También se pondera la justificación de los biomodelos empleados en los estudios preclínicos de esta enfermedad. De igual forma, se describen los antecedentes para medir las alteraciones, las técnicas y los métodos directos e indirectos de medición de la presión arterial, la cual fue provocada experimentalmente en los animales de laboratorio para realizar los estudios de hipertensión humana.

Medical and biological sciences require, as a priority, that research and experimentation be carried out on complete organisms (animal models). Its use has allowed the development of innumerable preclinical tests to evaluate pathogenic and therapeutic mechanisms of various diseases, as well as to study the causes, nature and cure of multiple human health disorders. In this work, we show a general overview of arterial hypertension biomodels where concepts, characteristics, origin, importance, utility and experimental procedures during their induction phase are described. The justification of the biomodels used in preclinical studies of this disease is also considered. Antecedents are also described to measure alterations, techniques and direct and indirect methods of measurement of arterial pressure, which was provoked experimentally in the laboratory animals to carry out the studies of human hypertension.

Effect of Cissus quadrangularis on Fracture Healing in Laboratory Animal / Efeito do Cissus quadrangularis na cicatrização de fraturas em animais de laboratório

Objectives: The present study was designed to examine the efficacy of Cissus quadrangularis paste on fracture healing in artificially induced fractured rabbits. Methods: Fifteen rabbits were separated into three groups namely A, B, and C. Veldt grape paste was applied in groups B and C (Treatment Group) by close reduction and open reduction methods of fracture management; respectively, while group A was kept as control. The blood parameter and fracture healing properties in all animals have been monitored and examined routinely during the study period. Results: Both treated groups revealed lower serum calcium levels (SCL) than the control group after 24 hours of fracture that became within the normal range on the 14th day. Fracture healing in the treated groups has been commenced more rapidly than the control group with complete bridging of discontinuity by a distinct osseous callus in the fracture line on day 7 and complete effacing of fracture line on day 14. Conclusion: We did not find any type of anomalousness, clinical deviations, and alteration of serum calcium level on the 14th day of the fracture in treated animals hence Veldt Grape paste could be readily applicable to the management of the fracture in animals.

Objetivos: avaliar a eficácia da pasta de Cissus quadrangularis na consolidação de fraturas em coelhos fraturados artificialmente. Metodos: quinze coelhos foram separados em três grupos (A, B e C). A pasta foi aplicada nos grupos B e C (Grupo de Tratamento) pelos métodos de redução fechada e redução aberta de gerenciamento de fraturas; respectivamente. O grupo A foi mantido como controle. O parâmetro sanguíneo e as propriedades de cicatrização de fraturas em todos os animais foram monitorados e examinados, rotineiramente, durante o período do estudo. Resultados: ambos os grupos tratados revelaram níveis séricos de cálcio (SCL) mais baixos do que o grupo controle, após 24 horas de fratura que se tornou normal no 14º dia. A cicatrização da fratura nos grupos tratados foi iniciada mais rapidamente do que o grupo controle, com ponte completa de descontinuidade por um calo ósseo distinto na linha de fratura no dia 7 e apagamento completo da linha de fratura no dia 14. Conclusao: não encontramos nenhum tipo de anomalia, desvios clínicos e alteração do nível sérico de cálcio no 14º dia da fratura nos animais tratados, portanto, a pasta Veldt Grape pode ser aplicável ao manejo da fratura em animais.

Efectos de la inyección neonatal de estreptozotocina sobre indicadores bioquímicos y de estrés oxidativo en ratas / Effects of neonatal injection of streptozotocin on biochemical indicators and oxidative stress in rats

Introducción: Debido a sus propiedades químicas, la estreptozotocina es uno de los agentes diabetogénicos más utilizados para generar modelos biológicos de diabetes, por lo que es necesario estudiar cuáles son sus efectos en el organismo del animal de laboratorio. Objetivo: Evaluar, en un periodo de 90 días, los efectos de la inyección neonatal de estreptozotocina en ratas Wistar sobre indicadores bioquímicos y de estrés oxidativo en hígado y riñón. Métodos: La diabetes fue inducida neonatalmente por 100 mg de estreptozotocina en ratas Wistar. Se realizaron determinaciones de glucemia, insulina e indicadores de estrés oxidativos en hígado y riñón en cinco animales por grupo a los días 5, 10, 20, 30, 60, 90 de nacidos. Resultados: En todas las intervenciones, la glucemia e insulina mostraron diferencias significativas en el grupo-STZ respecto al control. El valor máximo de hiperglucemia se observó al quinto día. La concentración de nitratos y nitritos en hígado fue mayor que en riñón. En comparación con el grupo control, en el tejido hepático del grupo-STZ la concentración de nitratos y nitritos resultó significativamente superior los días 10-20. En todas las intervenciones se detectó consumo de glutatión reducido en ambos órganos. En el hígado de las ratas STZ no se demostró daño a lípidos ni proteínas; sin embargo, en riñón se detectó daño significativo en ambas biomoléculas al quinto día. Conclusiones: Tanto la citotoxicidad de la estreptozotocina neonatal como las concentraciones de glucosa e insulina inducidas repercutieron negativamente sobre los indicadores de estrés oxidativo estudiados en tejido hepático y renal(AU)

Introduction: Streptozotocin is currently one of the most used diabetogenic agents to generate biological models of diabetes due to its chemical properties, so it is necessary to study the consequences of STZ for the organism of the laboratory animal. Objective: To evaluate in a period of 90 days the effects of neonatal injection of streptozotocin in Wistar rats on biochemical indicators and oxidative stress in liver and kidney. Methods: Diabetes was induced neonatally by 100 mg of streptozotocin in Wistar rats. Blood glucose, insulin and oxidative stress indicators in liver and kidney were determined in 5 animals per group at days 5, 10, 20, 30, 60, 90 of birth. Results: Blood glucose and insulin showed significant differences in the STZ-group respect to the control group in all interventions. The maximum value of hyperglycemia was observed on day-5. The concentration of nitrates and nitrites in liver was higher than in kidney. In liver tissue of the STZ-group, this indicator was significantly higher on days 10-20 compared to the control. In all interventions, reduced glutathione consumption was demonstrated in the STZ-group compared to control in both organs. In the liver of STZ rats no lipid or protein damage was demonstrated. However, in the kidney, significant damage in both biomolecules was detected in the STZ-group on day-5. Conclusions: Neonatal streptozotocin cytotoxicity as well as induced glucose and insulin concentrations had a negative impact on oxidative stress indicators studied in liver and kidney tissue(AU)

Ethical considerations in animal research: the principle of 3r's

In the last century, progress in the knowledge of human diseases, their diagnosis and treatment have grown exponentially, due in large part to the introduction and use of laboratory animals. Along with this important progress, the need to provide training and guidance to the scientific community in all aspects related to the proper use of experimental animals has been indispensable. Animal research committees play a primary role in evaluating experimental research protocols, from their feasibility to the rational use of animals, but above all in seeking animal welfare. The Institutional Committee for the Care and Use of Animals (IACUC) has endeavored to share several relevant aspects in conducting research with laboratory animals. Here, we present and discuss the topics that we consider of utmost importance to take in the account during the design of any experimental research protocol, so we invite researchers, technicians, and undergraduate and graduate students to dive into the fascinating subject of proper animal care and use for experimentation. The main intention of these contributions is to sensitize users of laboratory animals for the proper and rational use of them in experimental research, as well as to disseminate the permitted and unpermitted procedures in laboratory animals. In the first part, the significance of experimental research, the main functions of IACUC, and the principle of the three R's (replacement, reduction, and refinement) are addressed.

Allogenic platelet-rich plasma in induced ulcers in rat's cornea / [Plasma rico em plaquetas alogênico em úlceras de córnea induzidas em ratos]

The objective in this study was to evaluate the clinic effect of applying allogenic platelet-rich plasma (PRP) heated or not, for treating cornea ulcers, including the dosage of PDGF-BB in the cornea. The ulcers were induced, standardizing the left eye from 81 rats (Ratus norvegicus, albinus variety), assigned randomly into three groups (N=27): control group (CG) which did not receive any topic treatment; heated PRP group (GA) and PRP group (GP), which received topical treatment every eight hours for five days. Each group underwent evaluation at 24 hours (M1), three days (M3) and five days (M5). The clinical exam evaluated the opacity, vascularization and corneal repair. The corneal PDGF-BB was dosed through the ELISA method. The corneal opacity was decreased in PRP-treated animals (GA and GP) and corneal repair time reduced when compared to CG at M1 and M5. Furthermore, GP showed greater vascularization at M3 compared to M1. Applied allogenic PRP eye drops, heated or not, speed up corneal healing, and reduce corneal repair time. However, the corneal PDGF concentration was not altered in any of the treatments.(AU)

Objetivou-se avaliar o efeito clínico da aplicação de plasma rico em plaquetas alogênico (PRP) aquecido ou não, no tratamento de úlceras de córnea, como a dosagem de PDGF-BB na córnea. As úlceras foram induzidas, padronizando-se o olho esquerdo de 81 ratos (Rattus norvegicus, variedade albinus), aleatoriamente, nos três grupos (N = 27): grupo controle (CG), que não recebeu nenhum tratamento tópico; grupo PRP aquecido (GA) e grupo PRP (GP), que receberam tratamento tópico a cada oito horas, durante cinco dias. Cada grupo foi subdividido em 24 horas (M1), três dias (M3) e cinco dias (M5). O exame clínico avaliou a opacidade, a vascularização e o reparo corneano. O PDGF-BB corneano foi dosado pelo método Elisa. Houve diminuição da opacidade da córnea nos animais tratados com PRP (GA e GP) e diminuição do tempo de reparo da córnea em comparação com CG, M1 e M5. Além disso, foi observada maior vascularização no GP no momento M3 em relação ao M1. A aplicação de colírios de PRP alogênico, aquecidos ou não, acelera a cicatrização da córnea, além de reduzir o tempo de reparo da córnea. No entanto, a concentração de PDGF na córnea não se alterou em nenhum dos tratamentos.(AU)

Protocolo de prueba de ventilador mecánico para atención de pacientes con COVID-19 en modelo porcino / Mechanical ventilator test protocol for care of patients with COVID-19 in porcine model

RESUMEN A raíz de la pandemia generada por el SARS-CoV-2, los sistemas de salud de los distintos países han experimentado un dramático aumento en la ocupación de las unidades de cuidado intensivo y, por ende, una alta demanda de ventiladores mecánicos. En este artículo se propone un protocolo de pruebas de ventiladores mecánicos de bajo costo en modelos porcinos, como parte del desarrollo de equipos biomédicos que pueden apoyar la crisis sanitaria suscitada por la pandemia. El protocolo aborda aspectos que incluyen la fase previa a la prueba, el manejo anestésico, de vía aérea, seguimiento de laboratorio, recuperación y seguimiento del animal. Como resultado, se presenta a la comunidad académica un protocolo de validación de prototipo de ventilador mecánico en espécimen porcino como un referente para su aplicación y revalidación por parte de grupos interesados en el desarrollo de tecnologías locales de bajo costo.

ABSTRACT As a result of the SARS-CoV-2 pandemic, health systems globally have seen a dramatic increase in the occupancy of intensive care units, with mechanical ventilators being a resource in high demand in the care of these patients. This article proposes a protocol for testing low-cost mechanical ventilators in pig models, as part of the development of locally manufactured biomedical equipment that can support the health crisis caused by the pandemic. The protocol addresses aspects that include the pre-test phase, anesthetic and airway management, laboratory monitoring, recovery and monitoring of the animal. This document presents to the academic community a validation protocol of a mechanical ventilator prototype in a pig specimen that can be a reference for its application and revalidation by other groups interested in the development of local and low-cost technologies.

Protocolo de prueba de ventilador mecánico para atención de pacientes con COVID-19 en modelo porcino / Mechanical ventilator test protocol for care of patients with COVID-19 in porcine model

RESUMEN A raíz de la pandemia generada por el SARS-CoV-2, los sistemas de salud de los distintos países han experimentado un dramático aumento en la ocupación de las unidades de cuidado intensivo y, por ende, una alta demanda de ventiladores mecánicos. En este artículo se propone un protocolo de pruebas de ventiladores mecánicos de bajo costo en modelos porcinos, como parte del desarrollo de equipos biomédicos que pueden apoyar la crisis sanitaria suscitada por la pandemia. El protocolo aborda aspectos que incluyen la fase previa a la prueba, el manejo anestésico, de vía aérea, seguimiento de laboratorio, recuperación y seguimiento del animal. Como resultado, se presenta a la comunidad académica un protocolo de validación de prototipo de ventilador mecánico en espécimen porcino como un referente para su aplicación y revalidación por parte de grupos interesados en el desarrollo de tecnologías locales de bajo costo.

ABSTRACT As a result of the SARS-CoV-2 pandemic, health systems globally have seen a dramatic increase in the occupancy of intensive care units, with mechanical ventilators being a resource in high demand in the care of these patients. This article proposes a protocol for testing low-cost mechanical ventilators in pig models, as part of the development of locally manufactured biomedical equipment that can support the health crisis caused by the pandemic. The protocol addresses aspects that include the pre-test phase, anesthetic and airway management, laboratory monitoring, recovery and monitoring of the animal. This document presents to the academic community a validation protocol of a mechanical ventilator prototype in a pig specimen that can be a reference for its application and revalidation by other groups interested in the development of local and low-cost technologies.

Oogenic development and gonotrophic cycle of Aedes aegypti and Aedes albopictus in laboratory / Desarrollo oogénico y ciclo gonotrófico de Aedes aegypti y Aedes albopictus en laboratorio

Abstract: Objective: To determine the time of oogenic development and the length of the gonotrophic cycle of Ae. aegypti and Ae. albopictus in laboratory. Materials and methods: Bloodfed females of Ae. aegypti and Ae. albopictus were dissected every 4 h to determine the development status of the follicles according to the Christophers' stages. Results: The minimum time of oocyte maturation in Ae. aegypti and Ae. albopictus was 64-82 h and 52-64 h post-feeding, respectively. We found that the gonotrophic cycle of Ae. aegypti (3.7-4.2 d) is longer than that of Ae. albopictus (3.2-3.7 d). The follicle length showed significant differences between species at Christophers' stages 2" and 5, whereas follicle amplitude was different between the two mosquitoes at stages 2", 3 and 4. Conclusions: The study provided new evidence on the reproductive strategies of Ae. aegypti and Ae. albopictus females that coexist in the Neotropical region of Mexico.

Resumen: Objetivo: Determinar el tiempo de desarrollo oogénico y del ciclo gonotrófico de Aedes aegypti y Aedes albopictus en laboratorio. Material y métodos: Hembras de Ae. aegypti y Ae. albopictus alimentadas con sangre fueron disecadas cada cuatro horas para determinar el estado de desarrollo folicular, según los estadios de Christophers. Resultados: El tiempo mínimo de maduración del oocito en Ae. aegypti y Ae. albopictus fue de 64-82 h y 52-64 h post-alimentación, respectivamente. El ciclo gonotrófico de Ae. aegypti (3.7-4.2 d) fue mayor que el de Ae. albopictus (3.2-3.7 d). La longitud folicular presentó diferencias significativas entre las especies en los estadios de Christophers 2" y 5, mientras que la amplitud folicular fue diferente entre ambos mosquitos en los estadios 2", 3 y 4. Conclusiones: El estudio proporcionó nueva evidencia sobre la estrategia reproductiva de las hembras de Ae. aegypti y Ae. albopictus que coexisten en la región neotropical de México.