Cilostazol protects against changes caused by streptozotocin-induced diabetic retinopathy / Cilostazol protege contra as alterações causadas pela retinopatia diabética induzida por estreptozotocina

ABSTRACT Purpose: This study investigates the protective effect of cilostazol on the development and evolution of diabetic retinopathy in rats. Methods: Sixty male rats were divided into four groups: untreated nondiabetic rats, untreated diabetic rats, cilostazol-treated nondiabetic rats, and cilostazol-treated diabetic rats. The thickness of the internal limiting membrane to the outer limiting membrane, inner plexiform layer, inner nuclear layer, and outer nuclear layer were measured. The number of cell nuclei per 50-μm length in retinal sections was counted to quantify the degree of retinal cell loss. Results: The number of nuclei in the ganglion cell layer was significantly higher in untreated nondiabetic rats (p<0.05). The mean number of nuclei in the cilostazol-treated nondiabetic rats was significantly higher than that in the cilostazol-treated diabetic rats (p<0.05). The cilostazol-treated nondiabetic rats had a significantly higher mean nuclei count in the inner nuclear layer and inner plexiform layer as compared with the other groups (p<0.05). The total mean retinal thickness of the cilostazol-treated nondiabetic rats was significantly higher than that of cilostazol-treated diabetic rats and untreated diabetic rats (p<0.05). Conclusion: By decreasing the loss of ganglion cells and reducing the sensorineural atrophy in the internal retinal layers, cilostazol had a protective effect against changes caused by diabetic retinopathy in diabetic rats.

RESUMO Objetivo: O objetivo deste estudo foi investigar o efeito protetor do cilostazol no desenvolvimento e na evolução da retinopatia diabética em ratos. Métodos: Sessenta ratos machos foram divididos em 4 grupos: ratos não-diabéticos não-tratados, ratos diabéticos não-tratados, ratos não-diabéticos tratados com cilostazol e ratos diabéticos tratados com cilostazol. A espessura da membrana limitante interna à membrana limitante externa, a camada plexiforme interna, a camada nuclear interna e a camada nuclear externa foram medidas. Para quantificar o grau de perda de células da retina, foi contado o número de núcleos de células por 50 μm de comprimento em secções retinianas. Resultados: O número de núcleos no GCL foi significativamente maior em Ratos não-diabéticos não-tratados com cilostazol (p<0,05). O número médio de núcleos em Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol (p<0,05). A contagem média de núcleos em camada nuclear interna e camada plexiforme interna de ratos não-diabéticos tratados com cilostazol foi significativamente maior do que nos outros grupos (p<0,05). A espessura retiniana média total de Ratos não-diabéticos tratados com cilostazol foi significativamente maior do que em Ratos diabéticos tratados com cilostazol e Ratos diabéticos não-tratados (p<0,05). Conclusão: Os resultados demonstraram que o cilostazol teve um efeito protetor contra as alterações causadas pela retinopatia diabética em ratos diabéticos, diminuindo a perda de células ganglionares e reduzindo a atrofia neurossensorial nas camadas retinianas internas.

Therapeutic effect of transplantation of bone marrow mesenchymal stem cells co-cultured with bone marrow M2 macrophages on a rat model of liver cirrhosis / 临床肝胆病杂志

ObjectiveTo investigate the effect of transplantation of bone marrow mesenchymal stem cells （BMSCs） co-cultured with bone marrow-derived M2 macrophages （M2-BMDMs）， named as BMSCM2， on a rat model of liver cirrhosis induced by carbon tetrachloride （CCl4）/2-acetaminofluorene （2-AAF）. MethodsRat BMDMs were isolated and polarized into M2 phenotype， and rat BMSCs were isolated and co-cultured with M2-BMDMs at the third generation to obtain BMSCM2. The rats were given subcutaneous injection of CCl4 for 6 weeks to establish a model of liver cirrhosis， and then they were randomly divided into model group （M group）， BMSC group， and BMSCM2 group， with 6 rats in each group. A normal group （N group） with 6 rats was also established. Since week 7， the model rats were given 2-AAF by gavage in addition to the subcutaneous injection of CCl4. Samples were collected at the end of week 10 to observe liver function， liver histopathology， and hydroxyproline （Hyp） content in liver tissue， as well as changes in the markers for hepatic stellate cells， hepatic progenitor cells， cholangiocytes， and hepatocytes. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the N group， the M group had significant increases in the activities of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in ALT and AST （P<0.01）， and the BMSCM2 group had significantly better activities than the BMSC group （P<0.05）. Compared with the N group， the M group had significant increases in Hyp content and the mRNA and protein expression levels of alpha-smooth muscle actin （α-SMA） in the liver （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in Hyp content and the expression of α-SMA （P<0.05）， and the BMSCM2 group had a significantly lower level of α-SMA than the BMSC group （P<0.01）. Compared with the N group， the M group had significant increases in the mRNA expression levels of the hepatic progenitor cell markers EpCam and Sox9 and the cholangiocyte markers CK7 and CK19 （P<0.01） and significant reductions in the expression levels of the hepatocyte markers HNF-4α and Alb （P<0.01）； compared with the M group， the BMSC and BMSCM2 groups had significant reductions in the mRNA expression levels of EpCam， Sox9， CK7， and CK19 （P<0.05） and significant increases in the mRNA expression levels of HNF-4α and Alb （P<0.05）， and compared with the BMSC group， the BMSCM2 group had significant reductions in the mRNA expression levels of EpCam and CK19 （P<0.05） and significant increase in the expression level of HNF-4α （P<0.05）. ConclusionM2-BMDMs can enhance the therapeutic effect of BMSCs on CCl4/2-AAF-induced liver cirrhosis in rats， which provides new ideas for further improving the therapeutic effect of BMSCs on liver cirrhosis.

Postnatal Exposure to Exogenous progesterone Exacerbates the Prenatally-Induced Abnormal Sperms Production and Function in Rats / La exposición Posnatal a la Progesterona Exógena Exacerba la Producción y Función Anormales de Espermatozoides Inducidas Prenatalmente en Ratas

SUMMARY: This study aimed at clarifying the impact of long-term prenatal and postnatal exposure to exogenous progesterone on sperm production and function, relative sex organs weights, and the levels of the relevant hormones in rats. Sixty male Wistar rats were included and classified into three groups (n=20 in each). A test I group had mature rats born to dams treated with progesterone prenatally. A test II group included rats exposed to progesterone during prenatal as well as postnatal periods, and a control group had rats treated with a placebo (olive oil). The test groups revealed a significant reduction in sperm count, motility, and viability with higher abnormal forms than the control group (P< 0.05). Similarly, the test groups revealed significantly lower serum testosterone and higher FSH and LH levels (P< 0.001). Interestingly, the test II group showed pronounced sperm abnormalities, an alarming decrease in sperm viability and motility, and a significant accretion in the relative testicular weight compared to the test I group (p <0.001). Long-term (prenatal and early postnatal) treatment with synthetic progesterone hurts sperm quantity and quality, adversely affecting future male fertility.

Este estudio tuvo como objetivo aclarar el impacto de la exposición prenatal y posnatal a largo plazo a la progesterona exógena en la producción y función de los espermatozoides, el peso relativo de los órganos sexuales y los niveles de las hormonas relevantes en ratas. Sesenta ratas macho Wistar fueron incluidas y clasificadas en tres grupos (n=20 en cada uno). Un grupo de prueba I tenía ratas maduras nacidas de madres tratadas con progesterona prenatalmente. Un grupo de prueba II incluyó ratas expuestas a progesterona durante los períodos prenatal y posnatal, y un grupo de control tenía ratas tratadas con un placebo (aceite de oliva). Los grupos de prueba revelaron una reducción significativa en el recuento, la motilidad y la viabilidad de los espermatozoides con formas anormales más altas que el grupo de control (P < 0,05). De manera similar, los grupos de prueba revelaron niveles significativamente más bajos de testosterona sérica y niveles más altos de FSH y LH (P < 0.001). Curiosamente, el grupo de prueba II mostró anormalidades espermáticas pronunciadas, una disminución alarmante en la viabilidad y motilidad de los espermatozoides y una acumulación significativa en el peso testicular relativo en comparación con el grupo de prueba I (p <0.001). El tratamiento a largo plazo (prenatal y posnatal temprano) con progesterona sintética daña la cantidad y la calidad del esperma, lo que afecta negativamente la futura fertilidad masculina.

Sub-Chronic Safety Assessment on the Use of Solanum aethiopicum (L.) Leaf Extract as Blood Booster in Male Wistar Rats

Aim: The effect of Solanum aethiopicum (SA) on the haematological indices of Wistar rats was investigated in this study. Methodology: A total of 20 male Wistar rats with an average 172.45±0.15 g were distributed into four groups (A – D) and allowed to acclimatize for two weeks. Group A served as the control, while groups B, C, and D were given aqueous extracts of SA at doses of 75 mg/kg, 150 mg/kg, and 225 mg/kg per body weight, respectively, every 48 hours for 30 days. After the exposure period, a final evaluation and sacrifice of the rats was performed. Blood sample collection was carried for full blood count and blood film preparation. Results: The result of this study showed that leaf extract of Solanum aethiopicum caused a significant increase in white blood cells (18.18±0.78 - 27.08±2.68 x 103/?l), especially lymphocytes (13.58±2.48 - 30.95±4.65 x 103/?l) in group of rats when compared to control. On the contrary, there was a non-significant reduction in red blood cells (7.78±0.04 - 7.19±0.45 x 106/?l), hemoglobin (16.92±0.62 - 14.55±0.95 g/dl), haematocrit (41.49±0.29 - 38.38±1.68 %), mean corpuscular hemoglobin (21.71±0.91 - 20.30±0.10 ?g) when compared to the control. Platelet (451.25±87.25 - 724.75±249.25 x 103/?l) and Plateletcrit (0.36±0.07 - 0.50±0.17 %) was significantly higher in treated group, while mean platelet volume (8.21±0.31 - 6.98±0.07 ?m3) and platelet distribution width (18.68±1.38 - 15.93±0.73 %) was low when compared with control. Conclusion: The current study has demonstrated that the leaves of Solanum aethiopicum may be safe to consume in regulated amount, as it has been shown to boost blood indices. These plant extracts may be utilized as a blood promoting potentials as it has been shown to strengthens the body's immune system particularly cell-mediated immunity, have no hemotoxic impact on the red blood cell and its indices and improve the ability for the body to repair itself as seen from the platelet count and its indices.

Jatropha tanjorensis Euphorbiaceae Ethanolic Leaves Extract Reverses Phenylhydrazine Induced Haematological Alterations in Albino Wistar Rats

Introduction: Phenylhydrazine has been used in many studies to evaluate its modulatory effects in various biochemical parameters in whole blood and red blood cell lysate. Jatropha tanjorensis Euphorbiaceae have high antioxidants properties; its leaves phytochemical analysis shows the presence of flavonoids, tanins, terpenoids, saponis. This study investigated the ameliorative effects of Jatropha tanjorensis Euphorbiaceae on phenylhydrazine induced haematological alterations in albino Wistar rats. Materials and Methods: Wistar rats of both sexes (180-200g) were divided into 4 groups (n=5). Group 1 received rat chow; Group 2 received (200 mg/kg) of J. tanjorensis orally. Group 3 received phenylhydrazine only (10 mg/kg). Group 4 received phenylhydrazine (10 mg/kg) + J. tanjorensis (250 mg/kg). All animals were allowed free access to clean drinking water and normal rat chow ad libitum for 35 days. After which animals were sacrificed and blood samples collected for biochemical analysis. Results: Results obtained showed that phenylhydrazine induced normochromic anemia with significant increase in white blood cell count, and neutrophil counts, eosinophils (insignificant) count with a significant reduction in lymphocyte count. However, J. tanjorensis extract reversed the adverse haematological changes induced by phenylhydrazine. Conclusion: In conclusion, Jatropha tanjorensis Euphorbiaceae demonstrated antioxidant, anti-inflammatory, and anti-thrombotic effects and reversed the haematological alterations brought upon by phenylhydrazine administration.

Efecto regenerador gástrico del consumo de Petroselinum sativum L. (perejil) en ratas con gastritis inducida por etanol

El objetivo del presente trabajo fue determinar el efecto regenerador gástrico del consumo de Petroselinum sativum L. (perejil) en ratas con gastritis inducida por etanol. Se realizó un estudio analítico, experimental clásico, transversal, prospectivo. Se trabajó con 36 ratas Wistar machos (250 ± 30 g.p.c.) distribuidas aleatoriamente en 6 grupos (n=6). Los grupos II-VI fueron sometidos a ayuno de 24 horas para inducirles úlcera gástrica administrándoles 10 mL/kg.p.c. de etanol al 70% vía orogástrica. Después de una hora, se procedió a sacrificar al grupo II para observar el daño ulceroso en el estómago. Después, se elaboró el extracto acuoso de hojas frescas de perejil (EAHP) y se administró a los demás grupos el siguiente tratamiento por vía orogástrica durante 3 días: grupo III, 10 mL/kg.p.c. de solución de NaCl al 0,9%; y EAHP a los grupos IV-VI (150, 300 y 600 mg/kg.p.c., respectivamente). Enseguida, las ratas fueron sometidas a ayuno de 24 horas para luego ser sacrificadas por desnucamiento. Posteriormente, se les realizó una laparotomía para la extracción del estómago. El EAHP generó mayor producción de moco gástrico en las dosis de 300 mg/kg.p.c. con 78,03% y de 600 mg/kg.p.c. con 80,52% (p<0,05). Esto concordó con lo observado histológicamente en la mucosa gástrica, mostrando solo signos de inflamación de la submucosa en los grupos que consumieron EAHP (IV-VI), en comparación con necrosis fibrinoide de los grupos que no lo consumieron (II y III). En conclusión, el consumo de EAHP tiene un efecto regenerador gástrico en ratas con gastritis inducida por etanol.

Our objective is to determine the gastric regenerative effect of Petroselinum sativum L. (parsley) consumption in rats with ethanolinduced gastritis. We developed an analytical, experimental, classical, cross-sectional, prospective study. We worked with 36 male Wistar rats (250 ± 30 g.p.c.) randomly distributed into 6 groups (n=6). Groups II-VI were subjected to a 24-hour fast to induce gastric ulcer by administering 10 mL/kg.p.c. of 70% ethanol via orogastric. After one hour, group II was sacrificed to observe the ulcerative damage in the stomach. Afterward, the aqueous extract of fresh parsley leaves (EAHP) was prepared, and the following treatment was administered to the other groups through the orogastric route for 3 days: group III, 10 mL/kg.p.c. 0.9% NaCl solution; and EAHP to groups IV-VI (150, 300, and 600 mg/Kg.p.c., respectively). The rats were then fasted for 24 hours before being sacrificed by breaking their necks. Subsequently, a laparotomy was performed to extract the stomach. The EAHP generated greater production of gastric mucus in the doses of 300 mg/kg.p.c. with 78.03% and 600 mg/kg.p.c. with 80.52% (p<0.05). This was consistent with what was observed histologically in the gastric mucosa, showing only signs of inflammation of the submucosa in the groups that consumed EAHP (IV-VI), compared with fibrinoid necrosis in the groups that did not consume it (II and III). In conclusion, the consumption of EAHP has a gastric regenerative effect in rats with ethanol-induced gastritis.

Histomorphological Observations in Reproductive Organs of Wistar Rats Administered with Aqueous Leaf Extract of Momordica charantia

Effort to explore the adverse effect of the plant, Momordica charantia on mammals has remained inadequate despite previous attempt by earlier investigators. More glaring is the paucity of information on the histomorphological effect of the plant’s aqueous leaf extract hence the need to determine possible alteration of tissue structures in reproductive organs of adult Wistar rats which may affect their functions. This study aimed at determining the effect of Momordica charantia aqueous leaf extract (MCALE) on the reproductive organs of experimental adult Wistar rats. Materials and Methods: Twenty five rats (male and female) weighing 180-200g were randomly divided into five (5) groups of five rats each. The experimental groups, A to D were fed on standard diet and administered with 100, 200, 400 and 800 mg/kg body weight /day of MCE orally using gavage for 30 days. Rats in the control groups were fed on standard diet and physiological saline orally. Organs were harvested, fixed in 10% neutral buffered formalin (ovaries) and Bouin’s fluid (testes), embedded in molten paraffin wax, sectioned with a rotary microtome and stained with the haematoxylin and eosin technique. Stained slides were examined using the Olympus microscope. Results: Sections of ovaries administered 100 mg/kg of the extract showed vesicular spaces in corpus luteum and enlarged blood vessels. Sections treated with 200 mg/kg revealed follicular cyst and mild vacuolation of zona granulosa. Sections of the ovaries administered 400 mg/kg revealed degenerative changes, follicular cyst, mild vacuolation and reduction of zona granulosa layer while those treated with 800 mg/kg showed severe vacuolation of the zona granulosa layer. Conclusion: Momordica charantia caused histomorphological changes in ovaries of Wistar rats which could cause hormonal imbalance and infertility in females. No histomorphological changes were observed in male testes.

Action of enfuvirtide on the pregnancy of albino rats ( Rattus norvegicus albinus, Rodentia, Mammalia ): biological assay and functional and histological analyses of exposed maternal-fetal organs

ABSTRACT Objective To assess the effects of enfuvirtide on pregnancy in albino rats and their fetuses. Methods Forty pregnant EPM 1 Wistar rats were randomly allocated into four groups: control (E) (distilled water twice/day), G1 (4mg/kg/day enfuvirtide), G2 (12mg/kg/day enfuvirtide), and G3 (36mg/kg/day enfuvirtide) groups. On the 20th day of gestation, the rats were anesthetized and subjected to cesarean section. Their blood was collected for laboratory analysis, and they were sacrificed. The offspring's fragments of their kidneys, liver, and placentas and the maternal rats' fragments of their lungs, kidneys, and liver were separated in the immediate postpartum period for light microscopy analysis. Results No maternal deaths occurred. In the second week at the end of pregnancy, the mean weight of the G3 Group was significantly lower than that of the G2 Group (p=0.029 and p=0.028, respectively). Analyzing blood laboratory parameters, the G1 Group had the lowest mean amylase level, and the G2 Group had the lowest mean hemoglobin level and the highest mean platelet count. In the morphological analysis, there were no changes in organs, such as the kidneys and liver, in both the maternal rats and offspring. Three maternal rats in the G3 Group had pulmonary inflammation in the lungs. Conclusion Enfuvirtide has no significant adverse effects on pregnancy, conceptual products, or functional alterations in maternal rats.

Articular mobilization promotes improvement in functional and inflammatory parameters in a gouty arthritis model

ABSTRACT Objective Gouty arthritis is characterized by painful inflammation due to the deposition of monosodium urate crystals in joint tissues. Despite available treatments, many patients experience ineffective management and adverse effects. This study evaluated a manual therapy protocol involving passive joint mobilization at the peak of inflammation in a gouty arthritis model using functional and inflammatory parameters. Methods Twenty male Wistar rats, 12 weeks old, were divided into two groups (n=10 each): Gouty Arthritis and Control Groups, which were further subdivided into treated and untreated groups (n=5 each). The Gouty Arthritis Group received intraarticular knee injection of 50µL of monosodium urate crystals, while the Control Group received 50µL of phosphate buffered saline. The treatment involved a 9-minutes session of grade III joint mobilization (according to Maitland). Nociception, grip strength, and edema were evaluated before induction (EV0), 7 hours after assessment (EV1), immediately after treatment (EV2), and 1 hour after treatment (EV3). The animals were euthanized, and synovial fluid was collected to analyze leukocyte migration. Results The model mimicked the signs of the Gouty Arthritis Group, with a decrease in the threshold of nociception and strength and an increase in edema and leukocyte count. The mobilization protocol significantly increased the nociceptive threshold and grip strength and reduced edema; however, it did not reverse the increase in leukocyte count. Conclusion Our results suggest that mobilization promotes analgesia and may modulate the inflammatory process owing to reduced edema and subtle attenuation of cell migration, which contributes to strength gain.

Indicadores morfométricos de la corteza cerebral frontal de gazapos de ratas Wistar con diabetes pregestacional

Introducción: La diabetes es una enfermedad que afecta el embarazo provoca complicaciones fetales; dentro de ellas son frecuentes las malformaciones congénitas. Por la imposibilidad práctica y ética de estudiar este proceso en gestantes es imprescindible realizar estudios experimentales empleando procedimientos morfométricos para determinar si la diabetes afecta el neurodesarrollo. Objetivo: Caracterizar morfométricamente la sustancia gris de gazapos de ratas Wistar normales y con diabetes mellitus pregestacional. Métodos: Se realizó un estudio experimental básico de serie de casos a 20 gazapos de ratas Wistar de los cuales 10 eran descendientes de diabetes pregestacional. Se caracterizaron indicadores morfométricos del tejido nervioso como espesor de corteza e indicadores nucleares como el perímetro. Resultados: La media de la altura de la sustancia gris cortical mostró un valor de 1,224 ± 303,7 μm para el grupo control y 1,014 ± 376,0 para los casos, al aplicar el test de diferencias de medias se encontró diferencia significativa (p ≤ 0,05) a favor del grupo control. Los valores de la media del perímetro nuclear en el grupo control fue de 42,80 ± 7,23 μm y en el grupo experimental el promedio fue de 39,68 ± 6,52 μm, al aplicar el test de diferencias de medias se encontró diferencia significativa (p ≤ 0,05) a favor del grupo control al presentar mayor perímetro nuclear. Conclusiones: El mayor espesor cortical y perímetro nuclear correspondió al grupo control evidenciándose el efecto deletéreo de la diabetes mellitus en el neurodesarrollo.

Introduction: Diabetes is a disease that affects pregnancy causing fetal complications; within them congenital malformations are frequent. Due to the practical and ethical impossibility of studying this process in pregnant women, it is essential to carry out experimental studies using morphometric procedures to determine if diabetes affects neurodevelopment. Objective: To characterize morphometrically the gray matter of kits from normal Wistar rats and those with pregestational diabetes mellitus. Methods: A basic experimental study of a series of cases was carried out on 20 young Wistar rats, of which 10 were descendants of pregestational diabetes. Morphometric indicators of the nervous tissue were characterized as thickness of the cortex and nuclear indicators such as perimeter. Results: The average height of the cortical gray matter showed a value of 1.224 ± 303.7 μm for the control group and 1.014 ± 376.0 μm for the cases. When applying the mean difference test, a significant difference was found (p ≤ 0.05) in favor of the control group. The values of the measurement of the nuclear perimeter in the control group was 42.80 ± 7.23 μm and in the experimental group the average was 39.68 ± 6.52 μm. When applying the mean different test, a significant difference was found (p ≤ 0.05) at favor of control group presenting greater nuclear perimeter. Conclusions: The greatest cortical thickness and nuclear perimeter corresponded to the control group, evidencing the deleterious effect os diabetes mellitus on neurodevelopment.

Obesity influences the development of bisphosphonate-induced osteonecrosis in Wistar rats

Abstract Medication-related osteonecrosis of the jaw (MRONJ) is characterized by bone exposure for more than eight weeks in patients who have used or been treated with antiresorptive or antiangiogenic drugs, without a history of radiation therapy or metastatic diseases in the jaws. Obesity is associated with changes in periodontal tissues and oral microbiota that are linked to bone alterations. This study aimed to analyze the influence of obesity on the development of bisphosphonate-induced osteonecrosis. The experiment randomly and simply divided 24 male Wistar rats (Rattus norvegicus) into four groups: healthy, with osteonecrosis, obese, and obese with osteonecrosis (n=6 per group). Osteonecrosis was induced through weekly intraperitoneal injection for eight weeks at a dose of 250 µg/kg of zoledronic acid in a 4 mg/5 mL solution, combined with trauma (exodontia). Obesity was induced through a high glycaemic index diet. Each group was qualitatively and quantitatively evaluated regarding the development of models and pathological anatomy of the lesions. The results were expressed in mean percentage and standard deviation and statistically analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test, with a significance level of 5% (p<0.05) to establish differences found between the groups. Animals in the osteonecrosis group and the obese with osteonecrosis group presented larger necrosis areas (averages: 172.83±18,19 µm2 and 290.33±15,77 µm2, respectively) (p<0,0001). Bone sequestration, hepatic steatosis, and increased adipocyte size were observed in the obese group (average: 97.75±1.91 µm2) and in the obese with osteonecrosis group (average: 98.41±1.56 µm2), indicating greater tissue damage in these groups (p<0,0001). All parameters analyzed (through histological, morphometric, and murinometric analyses) increased for the obese and obese with osteonecrosis groups, suggesting a possible influence of obesity on the results. However, further studies are needed to confirm the role of obesity in the possible exacerbation of osteonecrosis and understand the underlying mechanisms.

Rattus norvegicus infectados com Trypanosoma cruzi: efeitos clínicos do tratamento com Lycopodium clavatun e phosphorus ultradiluídos / Rattus norvegicus infected with Trypanosoma cruzi: clinical effects of treatment with highly diluted Lycopodium clavatun and phosphorus / Rattus norvegicus infectado con Trypanosoma cruzi: efectos clínicos del tratamiento con Clavatun licopodio altamente diluido y fósforo

A doença de Chagas causada pelo parasita Trypanosoma cruzi acomete milhões de pessoas no mundo e não conta com um medicamento de ação efetiva para o seu tratamento etiológico. As drogas disponíveis, o nifurtimox e o benznidazol possuem índices de cura baixos com efeitos colaterais e toxidade que dificultam a adesão dos pacientes à terapia. Este fato impulsiona a busca por alternativas de tratamento que sejam mais efetivas e menos agressivas. Sendo assim, este trabalho teve como objetivo a avaliação dos efeitos clínicos apresentados por Rattus norvergicus infectados por T. cruzi e tratados com soluções ultradiluídas de Lycopodium clavatum ou Phosphorus. O estudo envolveu 93 ratos com quarenta e cinco dias de idade infectados intraperitonealmente com 5x106 formas tripomastigotas sanguíneos da cepa Y de T. cruzi, distribuídos nos grupos: Sadio SD (n=13) - controle não infectado e não tratado, grupo CI (n=27) - controle infectado e tratado com solução hidroalccólica 7% (etanol ­ água), grupo LY diluição 1:1x1026 (n=27) - infectado e tratado com Lycopodium, grupo PH diluição 1:1x1026 (n=26) - infectado e tratado com Phosphorum. Os animais foram avaliados clinicamente através dos parâmetros peso, temperatura, consumo de água e ração, quantidade de excretas, diâmetro e comprimento intestinal, aspecto da pelagem e consistência das fezes. Este estudo mostrou que os parâmetros utilizados foram importantes para a definição clínica da infecção de Rattus novergicus, linhagem Wistar pelo T. cruzi. Mostrou que os medicamentos LY e PH apresentam efeitos benéficos na evolução da clínica dos animais tratados. A utilização de Lycopodium clavatum e Phosphorus diluídos na proporção de 1:1x1026, apresentaram efeitos diferentes. Oito e seis parâmetros de quatorze analisados mostraram efeitos positivos para LY e PH, respectivamente. Os parâmetros consumo de água e ração, quantidade de excretas, diarreia, alopecia difusa e comprimento intestinal apresentaram diferenças significativas em relação ao controle infectado mostrando que mais estudos são necessários com o uso de medicamentos ultradiluídos na infecção pelo T. cruzi.

Chagas disease caused by the parasite Trypanosoma cruzi affects millions of people worldwide and does not have an effective drug for its etiological treatment. The available drugs, nifurtimox and benznidazole, have low cure rates with side effects and toxicity that make it difficult for patients to adhere to therapy. This fact drives the search for treatment alternatives that are more effective and less aggressive. Therefore, this work aimed to evaluate the clinical effects presented by Rattus norvergicus infected by T. cruzi and treated with ultradiluted solutions of Lycopodium clavatum or Phosphorus. The study involved 93 forty five day old rats intraperitoneally infected with 5x106 blood trypomastigotes forms of the Y strain of T. cruzi, distributed in the following groups: Healthy SD (n=13) - non-infected and untreated control, CI group (n =27) - infected control and treated with 7% hydroalcoholic solution (ethanol ­ water), LY group dilution 1:1x1026 (n=27) - infected and treated with Lycopodium, PH group dilution 1:1x1026 (n=26) - infected and treated with Phosphorum. The animals were clinically evaluated through the parameters weight, temperature, water and feed consumption, amount of excreta, intestinal diameter and length, coat appearance and feces consistency. This study showed that the parameters used were important for the clinical definition of infection of Rattus novergicus, Wistar lineage by T. cruzi. It showed that LY and PH drugs have beneficial effects on the clinical evolution of treated animals. The use of Lycopodium clavatum and Phosphorus diluted in the ratio of 1:1x1026, showed different effects. Eight and six parameters out of fourteen analyzed showed positive effects for LY and PH, respectively. The parameters water and feed consumption, amount of excreta, diarrhea, diffuse alopecia and intestinal length showed significant differences in relation to the infected control, showing that more studies are needed with the use of ultradiluted drugs in T. cruzi infection.

La enfermedad de Chagas causada por el parásito Trypanosoma cruzi afecta a millones de personas en todo el mundo y no cuenta con un fármaco eficaz para su tratamiento etiológico. Los fármacos disponibles, nifurtimox y benznidazol, presentan bajas tasas de curación con efectos secundarios y toxicidad que dificultan la adherencia terapéutica de los pacientes. Este hecho impulsa la búsqueda de alternativas de tratamiento más eficaces y menos agresivas. Por lo tanto, este trabajo tuvo como objetivo evaluar los efectos clínicos presentados por Rattus norvergicus infectados por T. cruzi y tratados con soluciones ultradiluidas de Lycopodium clavatum o Fósforo. En el estudio participaron 93 ratas de cuarenta y cinco días de edad infectadas intraperitonealmente con 5x106 formas tripomastigotes sanguíneas de la cepa Y de T. cruzi, distribuidos en los siguientes grupos: SD sano (n=13) - control no infectado y no tratado, grupo CI (n =27) - control infectado y tratado con solución hidroalcohólica al 7% (etanol - agua), grupo LY dilución 1:1x1026 (n=27) - infectado y tratado con Lycopodium, grupo PH dilución 1:1x1026 (n=26) - infectado y tratado con Phosphorum. Los animales fueron evaluados clínicamente mediante los parámetros peso, temperatura, consumo de agua y pienso, cantidad de excrementos, diámetro y longitud intestinal, aspecto del pelaje y consistencia de las heces. Este estudio demostró que los parámetros utilizados eran importantes para la definición clínica de la infección de Rattus novergicus, linaje Wistar por T. cruzi. Demostró que los fármacos LY y PH tienen efectos beneficiosos en la evolución clínica de los animales tratados. El uso de Lycopodium clavatum y Phosphorus diluidos en la proporción de 1:1x1026, mostró efectos diferentes. Ocho y seis parámetros de los catorce analizados mostraron efectos positivos para LY y PH, respectivamente. Los parámetros consumo de agua y pienso, cantidad de excretas, diarrea, alopecia difusa y longitud intestinal mostraron diferencias significativas en relación al control infectado, mostrando que son necesarios más estudios con el uso de fármacos ultradiluidos en la infección por

Moderate Toxicity of Potential Boron-containing Therapeutic, Dipotassium-trioxohydroxytetrafl uorotriborate - K2 (B3 O3 F4 OH) in Rats and Mice

Abstract Biological activity of boron-containing compounds (BCCs) has been well-known. Growing interest and numerous applications for BCCs have been reported. Boron and boron-containing acids show low acute toxicity in mammals but data on halogenated boroxine (HB) - dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH) acute toxicity have not been reported before. This compound, characterized as a potential therapeutic for skin changes, exhibits no observable genotoxicity in doses lower that 0.1 mg/ml in vitro and 55 mg/kg in vivo. It has also been confirmed as an antitumour agent both in vitro and in vivo as well as an inhibitor of enzymes involved in antioxidant mechanisms. The aim of this study was to assess the acute toxicity of HB and to determine the maximum tolerated dose as well as a dose free of any signs of toxicity in different test organisms. Acute toxicity of HB was tested in Sprague-Dawley and Wistar rats and BALB/c mice after single parenteral application of different doses. We determined doses free of any sign of toxicity and LD50 after single dose administration. LD50 of HB ranges from 63 to 75 mg/kg in different test models, meaning that HB shows moderate toxicity

Evaluation of the effect of intrathecal GM1 in 24, 48, and 72 hours after acute spinal cord injury in rats

Abstract Objective The aim of this study was to evaluate the best timing and feasibility of intrathecal application of sodium monosialoganglioside (GM1) after spinal cord contusion in Wistar rats as an experimental model. Methods Forty Wistar rats were submitted to contusion spinal cord injury after laminectomy. The animals were randomized and divided into four groups: Group 1 - Intrathecal application of GM1 24 hours after contusion; Group 2 - Intrathecal application of GM1 48 hours after contusion; Group 3 - intrathecal application of GM1 72 hours after contusion; Group 4 - Sham, with laminectomy and intrathecal application of 0.5 mL of 0.9% saline solution, without contusion. The recovery of locomotor function was evaluated at seven different moments by the Basso, Beattie, and Bresnahan (BBB) test. They were also assessed by the horizontal ladder, with sensory-motor behavioral assessment criteria, pre-and postoperatively. Results This experimental study showed better functional scores in the group submitted to the application of GM1, with statistically significant results, showing a mean increase when evaluated on known motor tests like the horizontal ladder and BBB, at all times of evaluation (p < 0.05), especially in group 2 (48 hours after spinal cord injury). Also, fewer mistakes and slips over the horizontal ladder were observed, and many points were achieved at the BBB scale analysis. Conclusion The study demonstrated that the intrathecal application of GM1 after spinal cord contusion in Wistar rats is feasible. The application 48 hours after the injury presented the best functional results.

Establishment of a rat model of acute radiation esophagitis / 中华放射肿瘤学杂志

Objective:To establish Wistar rat models of acute radiation esophagitis, and observe the histopathological changes at different time points after modeling.Methods:Wistar rats were locally irradiated with different doses of 6 MV X-rays, and the rats were sacrificed on the 3 rd, 5 th, 7 th, and 14 th days after irradiation. The full-length esophagus tissue was taken for paraffin embedding, sectioning, and hematoxylin and eosin (HE) staining for pathological assessment. The pathological changes of the esophagus of the rats were observed at the 3 rd, 5 th, 7 th, and 14 th days after 25 Gy and 30 Gy irradiation. The changes of daily dietary intake of rats in different irradiation groups within 1-2 weeks after radiation exposure were observed. Results:No rat died in two groups after being irradiated with 25 Gy and 30 Gy rays. All the rats in the 30 Gy group had esophagus injury. On the 7 th day, the degree of injury was the most serious, with a pathological score of 5.00±0.75 and a food intake of 0 g. On the 14 th day, the degree of injury was relieved, and the food intake was restored to the level before irradiation. Conclusions:The Wistar rat model of acute radiation esophagitis can be established by a single dose of 6 MV X-ray 30 Gy irradiation to the esophagus. The 7 th day after irradiation is an ideal observation time for the acute injury phase, which is gradually alleviated after the 7 th day. The time can be chosen from 7-14 days after irradiation as the observation point for the healing repair phase.

Daño en la pared arterial de las ratas Wistar sometidas a dieta hiperglucídica / Arterial wall damage in Wistar rats subjected to a hyperglycemic diet

RESUMEN Introducción: Los modelos experimentales en animales proporcionan una valiosa información para comprender los procesos fisiopatológicos de las lesiones de los vasos sanguíneos y sus consecuencias. Objetivo: Analizar los cambios histológicos y morfométricos que se observaron en la aorta abdominal de las ratas Wistar sometidas a una dieta hiperglucídica. Métodos: Se formaron aleatoriamente dos grupos experimentales de 10 animales cada uno. El grupo control alimentado con dieta estándar para la especie, y el grupo experimental alimentado con dieta estándar más sacarosa al 35 %, como agua de bebida desde el destete hasta las 20 semanas de vida. El estudio se realizó en muestras de aorta fijadas y procesadas por la técnica clásica de inclusión en parafina y coloreadas con las técnicas de hematoxilina - eosina y Verhoeff. Se realizó la descripción de las capas de la pared arterial y la determinación de variables morfométricas en cada lámina histológica. Resultados: Las ratas Wistar pertenecientes al grupo experimental desarrollaron modificaciones incipientes en la pared arterial de la aorta abdominal, las cuales corresponden con la presencia de tumefacción en la célula endotelial y vacuolización en la célula muscular lisa vascular, así como marcada desorganización de las fibras elásticas y musculares de la capa media. Las variables morfométricas que mostraron diferencias significativas entre los grupos fueron el grosor de la túnica media y el cociente media/ lumen. Conclusiones: La pared media resultó ser la capa más afectada demostrándose el efecto nocivo de la dieta hiperglucídica en la pared arterial.

ABSTRACT Introduction: experimental animal models provide valuable information to understand the physiological and pathological processes of blood vessel injuries and their consequences. Objective: to analyze the histological and morphometric changes observed in the abdominal aorta of Wistar rats subjected to a hyperglycemic diet. Methods: two experimental groups of 10 animals each were randomly formed. The control group was fed with a standard diet for this species, and the experimental group was fed with a standard diet plus 35% sucrose, as drinking water from weaning to 20 weeks of life. The study was performed on aortic samples fixed and processed by the classic paraffin embedding technique and stained with the hematoxylin-eosin and Verhoeff techniques. The description of the layers of the arterial wall and the determination of morphometric variables in each histological slide were made. Results: the Wistar rats belonging to the experimental group developed incipient changes in the arterial wall of the abdominal aorta, which correspond to the presence of endothelial cell swelling and vacuolation in the vascular smooth muscle cell, as well as marked disorganization of the muscle and elastic fibers of the middle layer. The morphometric variables that showed significant differences between the groups were the thickness of the tunica media and the media/lumen ratio. Conclusions: the middle wall turned out to be the most affected layer, demonstrating the harmful effect of the hyperglycemic diet on the arterial wall.

Comparative Evaluation of MTA Sealers Biomineralization Ability: an in Vivo Study / Avaliação Comparativa da Capacidade de Biomineralização de Cimentos MTA: Estudo in Vivo

This research aimed to evaluate the biomineralization induced by different concentrations of MTA Flow® and compare it to MTA Angelus®. Fifteen male Wistar rats received subcutaneous implants containing the materials to be tested (MTA Flow® at putty, thick, and thin consistencies and MTAAngelus®) and empty tubes (control). After 7, 40 and 90 days, the animals were euthanized, and the implants were removed with the surrounding tissue. The presence of biomineralization was analyzed in light microscope by Von Kossa technique. The statistica l differences were considered for p < 0.05. Calcification areas were present in all the MTA Flow® and MTA Angelus® groups. In the control group, no mineralized areas were observed. MTA Angelus® and thin-MTA Flow® showed significant reduction in calcification as time went by. A significant increase in areas with calcification, proportional to the exposure time, was observed in putty-MTA Flow® and thick-MTA Flow®. MTA Angelus® and thin-MTA Flow® showed significantly higher calcification than thick-MTA Flow® in the shortest exposure time. Analysis of putty-MTA Flow® showed significantly higher calcification areas than MTA Angelus® and thin-MTA Flow® in the longest exposure time. MTA Flow® stimulated mineralization, which has varied according to the concentration. Besides, in longer periods, MTA Flow® biomineralization performance was higher than MTAAngelus®, especially in highest concentration. (AU)

Esse estudo teve como objetivo avaliar a biomineralização induzida por diferentes concentrações do MTA Flow® e compará-la ao MTA Angelus®. Quinze ratos Wistar machos receberam implantes subcutâneos contendo os materiais a serem testados (MTA Angelus® e MTA Flow® nas consistências pastosa, espessa e fluida) e tubos vazios (controle). Após 7, 40 e 90 dias, os animais foram eutanasiados e os implantes foram removidos juntamente com o tecido circundante. A presença de biomineralização foi analisada em microscópio de luz pela técnica de Von Kossa. Diferenças estatísticas foram consideradas para valores de p < 0,05. Áreas de calcificação estavam presentes em todos os grupos do MTA Flow® e MTA Angelus®. No grupo controle não foram observadas áreas mineralizadas. O MTA Angelus® e o MTA Flow® fluido apresentaram redução significativa na quantidade de calcificação ao longo do tempo. Um aumento significativo na quantidade de áreas calcificadas, proporcional ao tempo de exposição, foi observado no MTA Flow® pastoso e no MTA Flow® espesso. O MTA Angelus® e o MTA Flow® fluido apresentaram calcificação significativamente maior do que o MTA Flow® espesso no menor período de exposição. Análises contento o MTA Flow® pastoso demonstraram áreas de calcificação significativamente maior do que o MTA Angelus® e MTA Flow® fluido no maior tempo de exposição. O MTA Flow® induziu a formação de áreas mineralizadas, que variou de acordo com a concentração do cimento. Em períodos mais longos, o MTA Flow® apresentou desempenho superior ao MTA Angelus®, principalmente quando utilizado na maior concentração. (AU)

Recording of motor and somatosensory evoked potential in an anaesthetised Wistar rat using digital polyrite system

Objectives: The aim of this article is to explain the detailed methodology to record Motor evoked potential (MEP) and somatosensory evoked potential (SSEP) in adult albino Wistar rat, male (200–250 g) which has not been defined previously. Materials and Methods: We have standardised recording of both MEP and SSEP in these rats under anaesthesia on ADI digital polyrite system. Results: Evoked potentials have been widely studied in spinal cord injured patients to estimate the degree of injury and to establish a predictive measure of functional recovery. MEPs and SSEPs, arising from the motor cortex or peripheral nerve and generated either by direct electrical stimulation or by transcranial magnetic stimulation, have been advocated as a reliable indicator of descending and ascending pathway integrity. In the rat brain, there is a physical overlap between the motor and somatosensory cortex. Hence, our objective was to identify the exact area for stimulation in the cortex where we could record maximum response with the application of minimum electrical stimulation. Conclusion: The recording of MEP and SSEP together provides a powerful neurological technique to monitor the tracts of the spinal cord.

A rat model of septic shock using live Escherichia coli and response to Noradrenaline

Objectives: Popular animal models of septic shock involve injections of endotoxin (bacterial lipopolysaccharide). Other methods that induce sepsis are often time-consuming and require long-term monitoring facilities. Further, individual models using different bacterial strains can deepen our understanding of sepsis pathophysiology. Hence, our objective was to develop an acute and functional Wistar rat model of septic shock using live strains of Escherichia coli and then administer Noradrenaline, a known sympathomimetic drug, to study if the response is along expected lines. Materials and Methods: After random allocation to one of three groups (Group 1 – E. coli alone, n=7; Group 2 – E. coli followed by Noradrenaline, n = 7 and Group 3 – control (n = 4), which received saline injections), Wistar rats were anesthetised and intra-arterial pressure was recorded from carotid artery catheter. Live E. coli suspended in normal saline (5 Mcfarland concentration; dose – 650 uL/100 g body weight) was injected through the tail vein to induce sepsis. When mean arterial pressure dropped to 50% of its value before E. coli injection, Noradrenaline was injected in Group 2. Results: The average time (t1, n = 14) for the septic shock to set in was about 1.94 ± 0.97 h. Six out of seven rats (Group 1) died within 60 min without intervention. The addition of Noradrenaline after hypotension in Group 2 prolonged the time to death significantly by about 170 min. Conclusion: The rat septic shock model using E. coli described in the study is an acute, stable, and functional model to study various aspects of septic shock. Administration of Noradrenaline prolonged the animal’s life in septic shock as expected. Future studies using other common sepsis agents encountered in clinics can be undertaken similarly

Relación del ácido fólico y los defectos del tubo neural en ratas Wistar / Relationship of folic acid and neural tube defects in Wistar rats

RESUMEN Introducción: los defectos del cierre del tubo neural son anomalías del sistema nervioso central superadas únicamente por los defectos cardíacos. En Pinar del Río existe un aumento de interrupciones de embarazo por diagnóstico de este defecto, se desconoce si los niveles de ácido fólico en las gestantes influyen en su incidencia. Objetivo: corroborar la relación entre los defectos del cierre del tubo neural en las crías de ratas Wistar y las dosis de ácido fólico administrada en las ratas Wistar gestadas. Métodos: se realizó un estudio experimental en ratas Wistar, se formaron cinco grupos de dos ratas hembras, dos que recibieron dosis de ácido fólico de 100 y 200 microgramos (µg) antes y durante la gestación, dos que recibieron 100 y 200 µg durante la gestación y un grupo control que no recibió ninguna dosis. La muestra estuvo constituida por 212 ratas recién nacidas, sacrificadas bajo normas internacionales. Se extrajo médula y cerebro para observar las anomalías, se tomaron fotomicrografía de los cortes realizados. El nivel de significación para todas las comparaciones estadísticas fue α ≤ 0,05. Resultados: se observaron en total trece espinas bífidas, once en el grupo control (26,19 %) y dos (4,65 %) en el de 200 µg durante la gestación, al comparar el grupo control respecto a los grupos tratados, la variable masa corporal presentaron significación estadística. Se observó que la presencia de espina bífida fue significativa en la hembra con respecto al macho. Conclusiones: se demostró la necesidad esencial de la ingestión de ácido fólico antes y durante la gestación en ratas Wistar y la asociación de espina bífida y sexo, el sexo femenino fue el predominante.

ABSTRACT Introduction: neural tube closure defects are anomalies of the central nervous system surpassed only by cardiac defects. In Pinar del Rio province there is an increase of pregnancy interruptions due to diagnosis of this defect, and it is not known if folic acid levels in pregnant women influence its incidence. Objective: to corroborate the relationship between neural tube closure defects in Wistar rats' pups and the doses of folic acid administered in pregnant Wistar rats. Methods: an experimental study was carried out in Wistar rats, forming five groups of two female rats; two of them received doses of folic acid of 100 and 200 micrograms (µg) before and during gestation, two that received 100 and 200 µg during gestation and a control group that did not receive any dose. The sample consisted of 212 newborn rats, sacrificed under international standards. Marrow and brain were extracted to observe the anomalies, and photomicrographs were taken of the sections made. The significance level for all statistical comparisons was α ≤ 0,05. Results: a total of thirteen spina bifida were observed, eleven in the control group (26,19 %) and two (4,65 %) in the 200 µg group during gestation, when comparing the control group with the treated groups, the body mass variable presented statistical significance. It was observed that the presence of bifid spina was significant in the female with respect to the male. Conclusions: the essential need of folic acid ingestion before and during gestation in Wistar rats and the association of spina bifida and sex were confirmed, being predominant in the female sex.