Subacute Administration Research Articles

Long Exposure to CdS-Dextrin Nanoparticles Induces an Immunomodulatory and Anti-Inflammatory Effect in Rats

Male Wistar rats were treated daily and intraperitoneally with CdS-Dextrin nanoparticles (100 μg/kg) during 30, 60, and 90 days. The effect of subacute and chronic administration of CdS-Dextrin nanoparticles on CD4/CD8 subpopulations of thymocytes, spleen-derived T cells, and peripheral blood CD4/CD8/CD3 T cells were analyzed by flow cytometer using a Rat T Lymphocyte Cocktail.

Sub-acute toxicity evaluation of ethanol extract of rheumatic tea formula (a polyherbal tea) in wistar rats

Sub-acute toxicity profile of Rheumatic Tea Formula (RTF), a polyherbal tea consisting of Salix alba, Eucalyptus globulus and Albizia chevalieri was investigated in wistar rats of both sexes. Wistar rats were orally administered three different doses of ethanol extract of RTF for 28 days after which the effect on body weight, feed consumption, water intake, liver and kidney functions as well as haematological parameters were evaluated. Also evaluated was the histolopathogical effect of the extract on the liver, kidney, spleen and stomach. The extract significantly (p 0.05) on kidney function indices was observed. There was also no significant effect (p > 0.05) on haematological parameters, except for WBC differentials where there was significant (p <0.05) elevation of lymphocytes and decrease in neutrophils. The extract was relatively non-toxic on kidney, liver, spleen, and stomach in the histopathological evaluations. The present study suggests that RTF extract might have no serious deleterious effect on sub-acute administration. Keywords : Rheumatic Tea Formula, sub-acute toxicity, liver, kidney, blood, wistar rats.

Apium graveolens extract influences mood and cognition in healthy mice.

Apium graveolens is a food flavoring which possesses various health promoting effects. This study investigates the effect of a sub-acute administration of A. graveolens on cognition and anti-depression behaviors via antioxidant and related neurotransmitter systems in mice brains. Cognition and depression was assessed by various models of behavior. The antioxidant system of glutathione peroxidase (GPx), % inhibition of superoxide anion (O2-), and lipid peroxidation were studied. In addition, neurochemical parameters including acetylcholinesterase (AChE) and monoamine oxidase-type A (MAO-A) were also evaluated. Nine groups of male mice were fed for 30days with different substances-a control, vehicle, A. graveolens extract (65-500mg/kg), and reference drugs (donepezil and fluoxetine). The results indicated that the effect of the intake of A. graveolens extract (125-500mg/kg) was similar to the reference drugs, as it improved both spatial and non-spatial memories. Moreover, there was a decrease in immobility time in both the forced swimming and tail suspension tests. In addition, the A. graveolens extract reduced lipid peroxidation of the brain and increased GPx activity and the % inhibition of O2-, whereas the activities of AChE and MAO-A were decreased. Thus, our data have shown that the consumption of A. graveolens extract improved cognitive function and anti-depression activities as well as modulating the endogenous antioxidant and neurotransmitter systems in the brain, resulting in increased neuronal density. This result indicated an important role for A. graveolens extract in preventing age-associated decline in cognitive function associated with depression.

Sub-acute administration of lower doses of nicotine caused sex-dependent improvement of renal function in Wistar rats.

The adverse and beneficial health effects of nicotine (NIC), the major alkaloid found in cigarettes and tobacco, are controversial. Most studies on NIC have focused on its effects on cardiovascular and nervous functions. This study aimed at determining dose- and sex-specific effects of sub-acute (28days) NIC administration on some indices of kidney function in Wistar rats. Forty rats (20 males and 20 females), 8-9 weeks old (each housed in separate metabolic cage), were used for this study such that graded doses of NIC (1, 2 and 4mg/kg i.p. for 28days) were administered to both sexes while each control received distilled water at 0.2mL/100g i.p. Blood was collected under ketamine anesthesia (10mg/kg i.m) for analyses and results obtained were compared at p<0.05. The result showed beneficial alterations in plasma and urine level of creatinine, urea and uric acid (p<0.05) as well as plasma and urine electrolyte level (Na+ and K+) in both sexes (p<0.05). Also, there was significant improvement in creatinine clearance (p<0.05) with no appreciable difference in their histological examination. Although these beneficial effects were more pronounced in the female than in the male (p<0.05), administration at the highest dose showed potentially deleterious alterations from normal beneficial trend (p<0.05) in both sexes. It was concluded that sub-acute administration of lower doses of NIC improves kidney function of Wistar rats; an effect that was more pronounced in the females than their male counterparts.

The Effects of Nanosilver, Encapsulated in a Polymeric Matrix, on Albino Rats Brain Tissue

We showed the results of immunohistochemical investigation of outbred albino rat’s neural tissue exposed to 9-day exposure of nanobiocomposite, which consist silver nanoparticles encapsulated in natural biopolymer matrix - arabinogalactan. Investigation of white rats was composed of 2 stages: half of the rats in each group were sacrificed immediately after exposure (early period) and other rats – through 6 months after end of exposure (distant period). We showed that test substance causes functional changes in nervous tissue. After subacute administration of nanobiocomposite - argentumarabinogalaktan (nAG) we have observed changes of content of apoptotic and anti-apoptotic proteins caspase-3 and bcl-2 in nervous tissue of white rats. number of normal neurons producing the protein caspase-3 was increased sharply and number of immunonegative normal neurons was significantly reduced. Along with this there was a high level of bcl – 2, one function of which is to prevent apoptosis trigger. We revealed in samples a significant increase in number of neurons with bcl-2, but protective effect of this protein not fully realized, which leads to significant increase of amount ofdamaged hyperchromatic cells. Assessment of results of immunohistochemical study of white rat’s nervous tissue, according to the expression of the protein caspase-3 and bcl – 2, may lead conclusion about ability of nanosilver encapsulated in a polymer matrix to penetrate blood-brain barrier and induced start apoptotic cascade in cortical neurons.

Effects of Aqueous Bryophyllum pinnatum Leaf Extract on Hematological, Renal and Sperm Indices in Wistar Rats

Bryophyllum pinnatum is a perennial herb, widely used in the treatment of several conditions in folklore medicine. The effects of sub-acute treatment of the aqueous extract of B. pinnatum leaf on hematological, renal and testicular functions were investigated in the Wistar rat. Adult rats were randomly distributed into three groups of six rats each and administered 0, 62.5 or 125 mg/kg of extract daily for 28 days by oral gavage. Blood levels of urea, creatinine and hematological parameters, as well as epididymal sperm parameters were measured. Kidney and testis of animals were removed and organ weight to body weight ratios were determined. Histology of the organs was also analyzed. The extract elevated white blood cell count, reduced neutrophil count (P<0.05) with out affecting lymphocyte count and packed cell volume, when compared to control. The extract did not alter sperm count, motility, morphology and viability as well as blood urea and creatinine levels. The extract did not alter histology and relative weights of the kidney and testis. It can be concluded that sub-acute administration of the aqueous extract of B. pinnatum did not produce any renal and testicular toxicity, but may depress neutrophil action.

Effects of Amlodipine on Spermatological Parameters and Genital Tract Weight in Adult Wistar Male Rats

Calcium ions are very important for many sperm functions involving sperm motility, reaction, and hyperactivation. The purpose of this study was to examine the effect of calcium channel blocker (CCB), Amlodipine (AML), on spermatological features, genital tract weight and spermatogenic cell density in adult male Wistar rats. Twelve rats (age, 12 weeks; weight, 290-350 g) were divided into two groups, six rats per each. Group I rats were fed normal diet without AML served as the control, Group II received 0.04 mg/daily AML for 30 days by oral gavage. Motility, left testis, right epididymis, prostate weight were decreased, and abnormal sperm rate was significantly (P<0.05) increased in AML groups compared with control group. The seminiferous tubular diameter and germinal cell layer thickness were not affected compared with control group. In conclusion, subacute administration of AML to male rats have detrimental effects on the spermatological parameters and genital tract weights, but not affect spermatogenic cell density of male rats.

Preclinical evaluation of anti-Helicobacter spp. activity of Hippocratea celastroides Kunth and its acute and sub-acute toxicity.

Background Hippocratea celastroides Kunth, commonly known as “cancerina”, is used in Mexican Traditional Medicine for the treatment of gastric and intestinal infections, systemic and skin inflammation, injuries and gastritis. The aim of this research was to assess the anti-Helicobacter pylori activities of hydro-ethanolic root-bark extracts from Hippocratea celastroides Kunth in naturally infected dogs, after testing their acute and subacute toxicities in mice.MethodsTo determine in vivo acute toxicity, a hydro-ethanolic extract was obtained and administered orally in female and male Balb-C mice, at doses ranging from 2000 to 5000 mg/kg. For the subacute study, a hydro-ethanolic extract was given to male and female Balb-C mice at doses ranging from 200 to 2000 mg/kg body weight. The animals were observed daily over a period of 42 days for signs of toxicity. In the pre-clinical anti-Helicobacter spp. assay, 60 dogs were included. Eighteen and 19 dogs for the experimental and control groups respectively, concluded the study. The experimental treatment consisted of H. celastroides hydro-ethanolic extract and the control treatment of amoxicillin-clarithromycin-omeprazole.ResultsOral LD50 (lethal dose 50) values for hydro-ethanolic extract were indeterminable at the highest tested doses. Under the subacute administration, neither mortality nor any sign of toxicity were observed when the hydro-ethanolic extract was administered. There were no significant alterations in biochemical parameters. The prevalence of Helicobacter spp. infection in dogs was 97.1 % for the experimental group and 100 % for the control group. Effectiveness was of 33.3 and 55 % in the experimental and control group respectively. The oral administration of H. celastroides was well-tolerated and safe.ConclusionThe root-bark of H. celastroides produced no signs of toxicity, and manifested pharmacological activity that indicated the possibility of an alternative treatment for H. pylori infection. Effectiveness is still low so it is necessary to continue research.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1412-6) contains supplementary material, which is available to authorized users.

Antidiabetic activity of two aqueous extracts of two cucurbitaceae: Citrullus colocynthis and Bryonia dioica

The aim of the present study was to evaluate the antidiabetic activity of the aqueous extract of Citrullus colocynthis L Schrad. (colocynth) fruits, CCFaq and Bryonia dioica Jacq. (White bryony) roots, BDRaq in neonatal streptozotocin-induced diabetic rats (n5-STZ diabetic rats). Experimental type 2 diabetes mellitus was induced by neonatal injection of streptozotocin (90 mg/kg i.p.) to pups of rats on the 5th day of birth. After 9 weeks, experimental type 2 diabetes mellitus was confirmed by polyphagia, polydipsia, polyuria, glucose intolerance, and fasting blood glucose levels > 1.50 g/L. Antidiabetic effect of CCFaq (20 mg/kg, i.p.) and BDRaq (30 mg/kg, i.p.) extracts as well as standard glibenclamide (10 mg/kg, i.p.) was evaluated on acute (3 h) and subacute (21 days) administration. Variations of body and some organs’ weight (pancreas, liver, kidney, and spleen) were recorded. Biochemical estimations of glucose, total cholesterol, triglycerides and urea levels were carried out during the experimentation. Intraperitoneal administration of CCFaq and BDRaq extracts as well as standard glibenclamide for 21 days to n5-STZ diabetic rats significantly (P < 0.01) decreased the level of serum glucose by –51%,–64% and–58% reductions, respectively. The body weight as well as serum levels of total cholesterol, triglycerides, and urea were markedly reversed secondary to subacute administration of aqueous extracts. In addition, changes in the weight of internal organs were restored to normal by the prolonged effect of BDRaq extract treatment. When compared with the CCFaq extract, the BDRaq extract appeared to have maximum antidiabetic activity in normalizing all studied parameters during the acute and subacute treatments. This study reveals the interesting antidiabetic effect of C. colocynthis and B. dioica and supports their traditional use for the treatment of diabetes mellitus.

Anti-hyperglycaemic and antioxidant effects of Bidens tripartita and quantitative analysis on its active principles.

Bidens species are used for their antidiabetic properties traditionally in many countries. Aim of this study is to evaluate hypoglycaemic and antidiabetic activity of Bidens tripartita extract and to identify its active compounds through bioactivity guided isolation technique. Hypoglycaemic effects of B. tripartita extract and its sub-extracts were investigated in normal and glucose-hyperglycaemic rats. Streptozotocin induced diabetic rats were used to examine antidiabetic activity of the extract and its sub-extracts after acute and sub-acute administration. Additionally, in vitro enzyme inhibitory and antioxidant activities were evaluated. HPLC analyses were carried out to determine the active constituents of the extract and its sub-extracts. Through in vivo bioactivity-guided fractionation process, ethyl acetate and n-buthanol sub-extracts were found to have potent antidiabetic activity. In vitro enzyme inhibitory activities of the same sub-extracts were found to be potent. The highest total phenol, flavonoid contents and radical scavenging activity was determined in ethyl acetate sub-extract. According to LC-MS analyses, chlorogenic acid, luteolin and 7-O-glucoside of luteolin (cynaroside) were determined as the main components of the active sub-extracts. According to our results, B. tripartita has potent antidiabetic activity and its active constituents might be beneficial for diabetes and its complications.

Safety evaluation (acute and sub-acute studies) of the aqueous extract of the leaves of Myrianthus arboreus P. Beauv. (Cecropiaceae) in Wistar rats

Ethnopharmacological relevanceMyrianthus arboreus P. Beauv (Cecropiaceae) is a medicinal plant distributed in forests and damp places of tropical Africa. Its leaves are widely used as food and/or for the treatment of various ailments including dysmenorrhoea, female infertility, tumors and diarrhea. However, to the best of our knowledge, no safety assessment of this plant has been reported yet. Aim of studyThe present study aimed at evaluating the safety of the aqueous extract of leaves of Myrianthus arboreus (MAA) in Wistar rats through an acute and sub-acute oral administration. Material and methodsIn acute oral toxicity, the test was performed according to the Organization for Economic Cooperation and Development (OECD) guidelines Nr. 423 (acute toxicity class method, ATC) with slight modifications. Female Wistar rats were orally treated with the aqueous extract of M. arboreus at the doses of 2000 and 5000mg/kg. In sub-acute toxicity study, using the OECD guidelines Nr. 407, the extract was administered by gavage at the doses of 20, 110 and 200mg/kg/day for 28 consecutive days. ResultsA single oral administration of 2000 or 5000mg/kg of the extract induced neither mortality nor exterior signs of toxicity indicating a LD50 >5000mg/kg. In sub-acute study, the extract decreased triglycerides, total cholesterol/high density lipoproteins ratio and atherogenic index of plasma in both sexes at all tested doses. Alanine transaminase decreased in both sexes at 200mg/kg and serum creatinine levels decreased at all tested doses in females. Moreover, significant increases in ovarian and uterine wet weights, red blood cell count, hematocrit, mean corpuscular hemoglobin and hemoglobin were observed at 200mg/kg in females. In males, this extract decreased white blood cell count, lymphocytes and relative weight of seminal vesicles and ventral prostate at 200mg/kg. ConclusionThe aqueous extract of Myrianthus arboreus leaves was non-toxic in acute administration and exhibited a relatively low toxicity potential on accessory sex organs in both sexes, and leukocytes in males following the repeated 28-days oral administration of the dose 200mg/kg.

Sub-Acute Toxicity Study of Tiger Milk Mushroom Lignosus tigris Chon S. Tan Cultivar E Sclerotium in Sprague Dawley Rats.

Lignosus also known as “Tiger Milk Mushroom,” is classified in the family Polyporaceae and mainly consumed for its medicinal properties in Southeast Asia and China. The sclerotium is known as the part with medicinal value and often used by the natives to treat a variety of ailments. Lignosus tigris Chon S. Tan, one of the species of the Malaysia Tiger Milk mushroom, has recently been successfully cultivated in laboratory. Earlier studies have demonstrated the L. tigris cultivar E sclerotia exhibited beneficial biomedicinal properties. This study evaluated the potential toxicity of L. tigris E sclerotia in a 28-day sub-acute oral administration in Sprague Dawley (SD) rats. L. tigris E sclerotial powder was administered orally at three different doses of 250, 500, and 1000 mg/kg to the SD rats once daily, consecutively for 28-days. Body weight of the rats was recorded and general behavior, adverse effects, and mortality were observed daily throughout the experimental period. At the end of the experiment, blood hematology and biochemistry, relative organ weights, and histopathological analysis were performed. Results showed that there were no mortality nor signs of toxicity throughout the 28-day sub-acute toxicity study. Oral administration of the L. tigris E sclerotial powder at daily dose up to 1000 mg/kg had no significant effects in body weight, relative organ weight, blood hematological and biochemistry, gross pathology, and histopathology of the organs. L. tigris E sclerotial powder did not cause any treatment-related adverse effect in the rats at different treatment dosages up to 1000 mg/kg. As the lethal dose for the rats is above 1000 mg/kg, the no-observed-adverse-effect level (NOAEL) dose is more than 1000 mg/kg.

STUDY ON HAEMATO-BIOCHEMICAL AND OXIDATIVE STRESS IN EXPERIMENTALLY INDUCED KETOPROFEN TOXICITY IN WISTAR RATS

The present research work was conducted to study the haemato-biochemical and oxidative stress effect of subacute intramuscular administration of ketoprofen for 14 days on 24 female Wistar rats. The animals were equally divided into 4 different groups A, B, C, D during randomization and dosed with vehicle control, 4 mg/kg as low dose, 8 mg/kg as mid dose and 12 mg/kg as high dose of ketoprofen, respectively. The drug was administered by intramuscular route every day for 14 days. At the end of 14 days treatment, all the animals from group A to D were subjected to blood collection and haemato-biochemical and oxidative stress investigation. The haematological study revealed dose dependent decrease in Hb, PCV,TEC, lymphocyte per cent and MCHC in Group C and Group D rats, while increase in MCV and neutrophil per cent in Group C and Group D rats.Administration of ketoprofen caused reduction in serum total protein (Group C and Group D), albumin (Group C and Group D) and globulin (Group D) in different groups. The result suggested that long term administration of ketoprofen at dose rate of 8 mg/kg and 12 mg/kg leads to macrocytic hypochromic anaemia possibly because of gastrointestinal bleeding or micro-bleeding and it is also evident that ketoprofen at the dose rate of 8 mg/kg b.wt. and 12 mg/kg b.wt. results into oxidative stress.

Anxiolytic-like effects and toxicological studies of Brickellia cavanillesii (Cass.) A. Gray in experimental mice models

Ethnopharmacological relevanceBrickellia cavanillesii (Asteraceae) (Cass.) A. Gray is one of the popular plants consumed in Central America and Mexico for the treatment of several diseases such as hypertension, diabetes and anxiety, among others. Aim of the studyTo determine the anxiolytic-like effect of B. Cavanillesii and the safety of its use through toxicological studies. Material and methodsAnxiolytic-like effects of soluble-methanol extract of B. cavanillesii (MEBc) were evaluated in ambulatory activity (open-field test), hole-board test, cylinder of exploration, the elevated plus-maze and the potentiation of the sodium pentobarbital-induced hypnosis mice models. On the other hand, in vivo toxicological studies were conducted on acute and sub-acute mice models recommended by OECD. Active MEBc was subjected to phytochemical studies through conventional chromatographic techniques to isolate bioactive compounds. ResultsMEBc (100mg/Kg) showed significant anxiolytic-like effect on animal model used (p<0.05). The phytochemical analysis of MEBc allowed the isolation of two major compounds nicotiflorin and acacetin, among others. Both compounds were found to be partially responsible for the anxiolytic-like effects. Moreover, a median lethal dose (LD50) higher than 2000mg/Kg was determined in mice and sub-acute oral administration of MEBc (100mg/Kg) did not alter body weight, clinical chemistry parameters (ALT and AST) and it did not induce any toxic nor alteration in the liver, kidney and heart functions. ConclusionsIn current investigation, we have shown that MEBc has a wide range of pharmacology–toxicology patterns. The results support further investigation of MEBc as a potential anxiolytic phytomedicinal agent.

Preclinical systemic toxicity evaluation of chitosan-solid&amp;ndash;lipid nanoparticle-encapsulated aspirin and curcumin in combination with free sulforaphane in BALB/c mice

Our previous studies have established the efficacy of chemopreventive regimens of aspirin and curcumin (CUR) encapsulated within solid lipid nanoparticles (SLNs) in combination with free sulforaphane (ACS combination) to prevent or delay the initiation and progression of pancreatic cancer, classified as one of the deadliest diseases with very low chances of survival upon diagnosis. Although toxicity of individual drugs and SLN has been studied previously, there are no studies in current literature that evaluate the potential toxicity of a combined regimen of ACS, especially when encapsulated within chitosan-SLNs (c-SLNs). Hence, objective of the current study was to investigate the potential toxic effects of ACS c-SLN combined chemopreventive regimens following acute (3 days), subacute (28 days), and subchronic (90 days) administrations by oral gavage in BALB/c mice. Mice were administered the following regimens: saline, blank c-SLN, low-dose ACS c-SLN (2+4.5+0.16 mg/kg), medium-dose ACS c-SLN (20+45+1.6 mg/kg), and high-dose ACS c-SLN (60+135+4.8 mg/kg). The potential toxicity was evaluated based on animal survival, body weight, hematology, blood chemistry, and organ histopathology. During 3-day, 28-day, and 90-day study periods, no animal deaths were observed. Treatment with ACS c-SLNs did not cause alteration in complete blood counts and blood chemistry data. Histopathological examination of various organ sections (pancreas, heart, liver, kidney, and brain) appeared normal. Based on the results of this study, no signs of toxicity in acute, subacute, and subchronic studies following oral administration of ACS c-SLNs were found indicating that the oral dosing regimens were safe at the levels tested for long-term administration to prevent the onset of pancreatic cancer.

Toxicological evaluation of colloidal nano-sized silver stabilized polyvinylpyrrolidone. IV. Influence on intestinal microbiota, immune indexes

Nano-sized colloidal silver (NCS) stabilized with polyvinylpyrrolidone (PVP) containing nanoparticles (NPs) of silver with a diameter of 10-80 nm was administered to growing male rats (body weight 80±10 g) during the first 30 days by intragastric gavage and then for 62 days with diet consumed in doses of 0.1, 1.0 and 10 mg/kg of body weight per day based on silver (Ag). The control animals received deionized water and PVP. The composition of microbiota from the cecum was studied using standard microbiological methods with determination of the main and transient components, together with antagonistic activity of symbiotic bifidobacteria. Expression of antigens CD45RA, CD3, CD4, CD8, CD161a on lymphocytes (Ly) of peripheral blood was determined by flow cytometry; blood serum levels of cytokines IL10, IL13, TNFα were examined by ELISA. It was shown that subacute administration of colloidal Ag in all studied doses did not lead to significant changes in the composition of the main components of normal microbiota, providing, however, the inhibitory effect on the growth of some transitory components probably including opportunistic species of microorganisms. Among the studied immunological parameters decreased amount of B-Ly was noticed at the highest dose of the NCS, while changes in the other parameters of the immune system were depended ambiguously on the dose of the product. The results were analyzed in conjunction with the data of previous publications concerning the impact on the NCS on integrated, morphological, hematological, biochemical and enzymological indexes of animals in the 92-day experiment. It was concluded that significant symptoms of NCS sub-acute oral toxicity manifested starting from a dose of 1 mg/kg body weight of Ag, and the maximum not observed adverse effect dose (NOAEL) can be estimated as 0.1 mg/kg body weight.

Sub-acute systemic erythropoietin administration reduces ischemic brain injury in an age-dependent manner.

Stroke is associated with neuroinflammation, neuronal loss and blood-brain barrier (BBB) breakdown. Thus far, recombinant tissue-type plasminogen activator (rtPA), the only approved treatment for acute ischemic stroke, increases the risk of intracerebral hemorrhage and is poorly efficient in disaggregating platelet-rich thrombi. Therefore, the development of safer and more efficient therapies is highly awaited. Encouraging neuroprotective effects were reported in mouse models of ischemic stroke following administration of erythropoietin (EPO). However, previous preclinical studies did not investigate the effects of EPO in focal ischemic stroke induced by a platelet-rich thrombus and did not consider the implication of age. Here, we performed middle cerebral artery occlusion by inducing platelet-rich thrombus formation in chimeric 5- (i.e. young) and 20- (i.e. aged) months old C57BL/6 mice, in which hematopoietic stem cells carried the green fluorescent protein (GFP)-tag. Recombinant human EPO (rhEPO) was administered 24 hours post-occlusion and blood-circulating monocyte populations were studied by flow cytometry 3 hours post-rhEPO administration. Twenty-four hours following rhEPO treatment, neuronal loss and BBB integrity were assessed by quantification of Fluoro-Jade B (FJB)-positive cells and extravasated serum immunoglobulins G (IgG), respectively. Neuroinflammation was determined by quantifying infiltration of GFP-positive bone marrow-derived cells (BMDC) and recruitment of microglial cells into brain parenchyma, along with monocyte chemotactic protein-1 (MCP-1) brain protein levels. Here, rhEPO anti-inflammatory properties rescued ischemic injury by reducing neuronal loss and BBB breakdown in young animals, but not in aged littermates. Such age-dependent effects of rhEPO must therefore be taken into consideration in future studies aiming to develop new therapies for ischemic stroke.

Acute and subacute toxicity studies of CMICE-013, a novel iodinated rotenone-based myocardial perfusion tracer, in Sprague Dawley rats and Gottingen minipigs

PurposeExtensive acute and subacute toxicities studies are required to evaluate the toxicological profile of the novel cardiac perfusion imaging tracer 123I-CMICE-013 to support applications for clinical trials. MethodsSprague-Dawley rats and Gottingen minipigs received injections of non-radioactive 127I-CMICE-013 at two dosage levels of 1 and 5 μg/kg, and vehicle buffer as control. In the acute toxicity studies, each animal was injected on two occasions 24 h apart and then underwent a 14-day recovery period; in the subacute study, animals received daily injections for 14 days continuously. The health status and mortality of test animals were monitored daily and body weight, food consumption, physiological and biochemical parameters were measured at various time points during the study. Animals were euthanized at the end of the studies and dissected for pathologic examination of organs and tissues. ResultsThe acute and subacute administrations of injections of the non-radioactive CMICE-013 in rats and minipigs were well tolerated. Little to no dosing-related adverse effects were observed in animal body and organ weights, hematology, coagulation, clinical chemistry, urinalysis, ophthalmoscopy, electrocardiograms, heart rates, blood pressure, macroscopic and microscopic examination of the preserved animal tissues including the brain. ConclusionThe lack of adverse effects from acute and subacute dosing suggest that the CMICE-013 injection solution has a reasonable safety margin within the designed concentration range to be utilized in imaging applications. The dosage level of 5 μg/kg was considered the no adverse effect level for both rats and minipigs based on our acute and subacute studies.

Anti-inflammatory and toxicological evaluation of Moussonia deppeana (Schldl. & Cham) Hanst and Verbascoside as a main active metabolite

Ethnopharmacological relevanceMoussonia deppeana, known as Tlachichinole, is a Mexican medicinal plant used for treatment of inflammatory diseases, influenza, diarrhea, gastrointestinal disorders and arthritis. Aim of the studyIn this paper the antioxidant and anti-inflammatory activities as well as the acute and sub-acute toxicological effects were evaluated for the ethanolic extract from aerial parts of M. deppeana, also its phytochemical analysis is described. Materials and methodsPhytochemical analysis and compound isolation were performed with thin layer chromatography. The chemical identification of the main compound was performed by 1H NMR (COSY, NOESY, HSQC and HMBC) spectra. In vitro antioxidant capacity and total phenolic content for the ethanolic extract and its primary fractions was determined by DPPH and Folin-Ciocalteu reagent. Acute and subacute toxicity tests were evaluated on Balb/C mice. Finally acute anti-inflammatory evaluation was tested for a local (TPA) and systemic (carrageenan) murine model. ResultsThe main compound isolated from the ethanolic extract of M. deppeana was Verbascoside, which was isolated from F3 and was identified by 1H NMR and COSY data. Furthermore oleanolic and ursolic acids were isolated from primary fractions F1 and F2. Ethanolic extract showed IC50 = 6.71mg/mL for DPPH test and 664.12µg QE/mL for the total phenolic content. The LD50 value was >2g/kg by i.g. route in male and female mice. Sub-acute administration (28 days) of the ethanolic extract (1g/kg) did not cause lethality or alter any hematological and biochemical parameters, in addition, histological analysis of the major organs exhibited no structural changes. Anti-inflammatory activity of the ethanolic extract showed an ED50 = 1.5mg/ear and 450mg/kg for TPA and carrageenan test, respectively. Primary fractions generated moderate local and systemic anti-inflammatory activity. ConclusionThe ethanolic extract from the aerial parts of M. deppeana did not cause any lethality or adverse effect in either of the acute and sub-acute toxicity tests. This exhibited an important local and systemic anti-inflammatory activity and also moderate antioxidant capacity. Moreover, the primary fraction F2 was more active for the TPA model while the primary fraction F3 was most active in the carrageenan model in vivo. The main compound isolated from F3 was verbascoside; on the other hand also ursolic and oleanolic acids were isolated from F1 and F2.

Effect of Sub-Acute Insulin Treatment on Short-Term Non-Spatial Working Memory in Mice

Insulin is well known as a trophic hormone that regulates glucose, protein and lipid metabolism in target tissues. It was, until recently, considered as a peripheral hormone that had no effect on the central nervous system. It is now well established that insulin occurs in the brain where it exerts many physiological effects. The aim of the study was to determine the effect of sub-acute insulin administration on non-spatial working memory in mice. Twelve mice of both sexes, weighing between 18 - 22 g, were divided into two groups and treated with either insulin or de-ionized water (control) for seven days. Short-term working memory was assessed using novel object recognition task. Time spent exploring the objects did not differ between the groups. Novel object recognition and discrimination ratio was also similar for the control and insulin-treated animals. It is concluded that sub-acute administration of insulin has no effect on short-term non-spatial working memory in treated mice.