Subacute Tests Research Articles

Toxicity and bacterial anti-motility activities of the hydroethanolic extract of Acacia senegal (L.) Willd (Fabaceae) leaves

BackgroundAcacia senegal is a plant traditionally used for its various properties, including the treatment of infectious diseases. Recently, our team has demonstrated the ability of the hydroethanolic extract of the leaves to increase the activity of phenicol antibiotics against multi-resistant bacteria. The aim of this work is to determine the toxicological effects of the extract and its capacity to inhibit the bacterial mobility of Gram-negative bacteria, in order to evaluate the level of safety use of this plant.MethodsThe cytotoxicity test was performed using the neutral red absorption method. Acute and sub-acute oral toxicity were conducted on NMRI mice and Wistar rats. The behaviour and adverse effects were recorded during the 14 days of the acute study. For the subacute test, biochemical parameters, food and water consumption, and morphological parameters were determined. The anti-motility activities were evaluated on Pseudomonas aeruginosa PA01 and Escherichia coli AG100, using specific concentrations of Agar as required by the method.ResultsHEASG induced inhibition of keratinocytes cell growth with an IC50 of 1302 ± 60 μg/mL. For the acute toxicity study in mice, the single dose of extract of 2000 mg/kg body weight caused no deaths and no behavioural changes were observed; therefore, the median lethal dose (LD50) of HEASG was calculated to 5000 mg/kg body weight. In Wistar rats, no mortality was observed at 250, 500 and 1000 mg/kg/day during the 28-day subacute oral toxicity study. The weights of both females and males increased globally over time, regardless of the batch. No statistically significant differences were registered for organ weights and biochemical parameters, except for chloride for biochemical parameters. Water and food consumption did not change significantly. Furthermore, no macroscopic changes in organ appearance were observed. Regarding anti-motility activity, the extract has reduced the swarming motility of PA01 and AG100 significantly at the concentration of 32 μg/mL (P < 0.001). The extract has reduced the swimming motility (P < 0.01) of PA01 but not AG100.ConclusionsThe results suggest that hydroethanolic extract of A. senegal leaves has significant activity against bacterial motility and relatively low toxicity.

Safety Evaluation of &lt;i&gt;Carex baccans&lt;/i&gt; Nees. (A Traditional Anthelminthic Remedy), in Swiss Albino Mice: An Acute and Sub-acute Toxicity Study

The medicinal properties of Carex baccans Nees. (Cyperaceae) has widely been exploited in traditional healing practices for the treatment of various ailments. Though, this is a widely used herb, not much is known about the adverse implications it may have. Therefore to shed some light into this aspect, acute and sub-acute toxicity study of the plant's crude extract and fractions were carried out in Swiss albino mice following Organization for Economic Co-operation and Development 423 and 407 guidelines respectively. Additionally, the phytochemical composition of the crude extract was also determined. Effect of the extract and the fraction was analyzed taking into consideration the body weight, organs weight, blood physiology, key markers of liver and kidney functions, and microscopic analysis of liver, kidney and intestine structural composition using scanning and transmission electron microscopy. The results showed that the crude extract exert no observable signs of acute toxicity even at a dose of 5000 mg/kg body weight of mice, however, the butanol fraction proved to be toxic with a mean lethal dose of 4677.35 ± 103.38 mg/kg. Repeated oral sub-acute test of the mice with crude extract and fraction of the crude extract was also found to exert adverse effects on the animals at high doses (500 mg/kg body weight and above) when compared to control group. These adverse implications were manifested in the body weight, hematology, biochemistry, and the structural and ultrastructural composition of some important organs. Therefore, it may be concluded that the plant C. baccans, if taken in its crude native form has a high safety margin at a single dose but not the butanol fraction. Furthermore, repeated exposure to the plant, both in its crude and fractionated (butanol fraction) form at high doses should be discouraged as it may have severe implications.

Research article Toxicity Studies on Kushta Khabasul Hadeed (calcined iron rust) prepared by Classical and Muffle Furnace Methods

&#x0D; Background: Though a number of studies has been carried out in terms of safety of Kushta but concerns have been raised about the presence of heavy metals in Unani formulations. Classical method of preparation of KKH and other Kushta preparation in Unani system of medicine has been found to eliminate all such issues from the end product. Hence; the present study was designed to evaluate the acute and sub-acute toxicity study of KKH prepared by Classical and Muffle furnace methods.&#x0D; Objective: To ascertain the safety profile of Kushta Khabasul Hadeed (KKH) prepared by classical and muffle furnace methods.&#x0D; Materials and methods:&#x0D; Both of the samples of KKH i.e. Kushta Khabasul Hadeed (KKHCM) and Kushta Khabasul Hadeed (KKHMFM) prepared by Classical and Muffle Furnace Methods respectively were evaluated for acute and sub-acute toxicity by OECD guidelines 423 and 407 respectively. The starting dose in the acute test was 50 mg/kg and in sub-acute test, it was 134mg, 200 mg and 400 mg/kg b.w. Effect of both of the test samples were assessed for ponderal changes, food and water intake, relative organs weights, hematological, biochemical and histopathological features of various organs.&#x0D; Results: Both of the samples did not produce any sign of toxicity in acute toxicity study up to the dose level of 2000mg/kg b.w. In sub-acute toxicity study no mortality in any of the test groups was observed. KKHCM at different doses showed a slight decrease in food intake in all the test groups but significant difference (P&lt;0.05) was observed in the test group B as compared to plain control.&#x0D; Conclusion: KKHMFM was found to be relatively toxic than KKHCM at different dose levels. The study demonstrated that both the Classical and Muffle furnace methods of preparation of Kushta are useful for preparing KKH and also safe at the dose prescribed in Unani literature. However, the Kushta prepared by the classical method was found to be safer than the muffle furnace method.

Effects of ammonia nitrogen on early growth and survival of the ‘Zebra 2’ strain of the manila clam ( Ruditapes philippinarum )

The ‘Zebra 2’ strain of the Manila clam (Ruditapes philippinarum) is a newly bred for aquaculture in the low temperature Bohai and Yellow seas in northern China. The goal of this study was to determine the tolerance of ‘Zebra 2’ clams at early developmental stages to total ammonia nitrogen (TAN) under acute stress and continuous subacute stress levels. ‘Zebra 2’ were cultured under TAN stress at water salinity of 27–28, temperature of 21–28°C and pH of 8.0–8.2. Survival rate and production of clams from the fertilized egg to juvenile stages were measured. The acute stress test revealed that the median effective concentrations of TAN for hatchability and deformity of fertilized eggs at 24 h were 7.08 and 15.12 mg/L, with the unionized ammonia (UIA) at 0.22 and 0.36 mg/L respectively. The 50% lethal concentrations for D-shaped larvae (shell length 101.50 μm), umbo larvae (179.67 μm) and juveniles (610.50 μm) at 96 h were 3.28, 48.90 and 57.57 mg/L, with UIA at 0.10, 2.08 and 2.99 mg/L respectively. These results suggested that the ‘Zebra 2’ clams hatching and D-shaped larvae stages were most sensitive to TAN and that the tolerance significantly increased after the umbo larval stage. In the continuous subacute stress test, ‘Zebra 2’ juveniles were observed under all the treatments after 53 days of exposure to TAN stress at 5.45 mg/L (0.23 mg/L UIA). The maximal acceptable toxicant concentration of UIA used to determine the mortality rate, yield and relative yield was ranged from 0.12 to 0.23 mg/L. These findings suggested that maintaining TAN <0.22 mg/L and UIA ≤0.021 mg/L at seawater pH ≤8.5, salinity ≥25 and water temperature ≤25°C would improve larval rearing of ‘Zebra 2’ clams.

Protective Effects of Garcinia kola Stem Barks Aqueous Extract on Metabolic Disorders and Oxidative Stress in Dexamethasone-Induced Insulin Resistance in Rat

Aim: To evaluate how the aqueous extract of Garcinia kola (GK) stem bark impacted dexamethasone-induced insulin resistance in male rats. Method: This work was completed in 2020. A phytochemical screening of the extract was first carried out. Using acute and subacute studies (11 days), the effect of the extract was evaluated at 100 mg/kg and 200 mg/kg, on dexamethasone-induced hyperglycemic rats. Glycemia was measured before and after treatment in both studies, while histological examination for isolated liver, kidneys and pancreas was performed, body weight and some internal organs determined, and biochemical assays. for blood samples were performed only for the subacute study. Results and Conclusion : An examination of the plant's chemistry showed the presence of alkaloids, phenolic compounds, flavonoids, saponins, coumarins, anthocyanins, and anthraquinones. GK extract decreased the Area Under Curve induced by dexamethasone (DEXA) injection both in the acute and in the subacute test. The extract and glibenclamide decrease the alanine amino transferase activity induced by DEXA. In both the glibenclamide and extract groups, total protein levels increased. The uric acid level in all dexamethasone-treated rats remains elevated when compared to the normal control group. Superoxide dismutase activity increased in both groups treated with the extract or glibenclamide. DEXA induced islet hypertrophy in the pancreas, vascular congestion, cytolysis, and vacuolation of hepatocytes. The kidney presented leucocyte infiltration and a decrease in glomerular cell density. All those abnormalities were improved by the administration of glibenclamide or the extract of Garcinia kola.

Safety assessment of essential oils from Xylopia aethiopica (Dunal) fruit from Nigeria

Introduction: The essential oils of Xylopia aethiopica fruits have been reported to possess numerous biochemical effects. However, no toxicological data is available regarding the safety evaluation of the essential oils from the plant spice. The present study was performed to evaluate the safety of essential oils from X. aethiopica (XAEO) fruit by acute and sub-acute oral toxicity studies in experimental rodents.&#x0D; Methods: In assessing the safety of XAEO, acute and sub-acute oral toxicity tests were performed following OECD guidelines 425 and 407, respectively, with slight modifications. At the end of each test, hematological and biochemical analysis of the collected blood was performed. Histopathological examination of vital organs of the animals was conducted for gross findings and for comparison to controls.&#x0D; Results: In acute oral toxicity, signs of toxicity were observed in the initial period of the experiment which culminated in the death of the mice before the end of the experiment. The sub-acute test observations indicated that generally, there were no significant differences (p&amp;lt;0.05) up to the high dose level compared to the controls.&#x0D; Conclusion: This study demonstrated the tolerability of XAEO administered daily for 28 days up to 450 mg/kg dose.

Acute, Sub-acute and Cell Toxicity of Allium elburzense Bulb Hydroalcoholic Extract

Background and objectives Allium elburzense is an endemic plant in north of Iran with some nutritional and medicinal applications; however, there is no data on its safety profile. This study was aimed to investigate cytotoxicity, acute and sub-acute toxicity of hydroalcoholic extract of A. elburzense bulb. Methods Total phenolic content of the extract was measured using Folin-Ciocalteumethod. For cytotoxicity assay, human umbilical vein endothelial cells (HUVECs) were used. In acute toxicity study, single oral dose of 2000 mg/kg was administered in female and male Wistar rats and they were monitored for two weeks. In sub-acute test, 125, 250 and 500 mg/kg/day of extract were orally administered for four weeks. Results Total phenolic content was estimated as 32.8 ± 2.5 mg gallic acid equivalent/g of the extract. The extract showed IC50 value of 366.4 µg/mL (95% CI = 246.4-566.1) in HUVECs after 24 h exposure. In acute study, there was no sign of toxicity and no mortality; however, significant increase in relative spleen weight and ALP activity and mild inflammation in kidney tissue were observed. LD50 ˃ 2000 mg/kg was estimated for A. elburzense bulb extract. In sub-acute assay, there were significant elevations in relative spleen weight, blood urea level, AST, ALT, ALP, total WBC, lymphocyte and neutrophil count and significant decrease in blood sugar and triglyceride levels at higher doses of the extract. ConclusionAllium elburzense bulb extract may be considered as safe at doses lower than 500 mg/kg in rats; however, assessment of liver and kidney functions is recommended during chronic uses.

Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test

Abstract Background Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. Results No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5 g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0 g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. Conclusions The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains. How to cite: Tsai WC, Chang HC, Tseng YH, et al. Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test. Electron J Biotechnol 2020;45. https://doi.org/10.1016/j.ejbt.2020.04.001

Assessment of the Knowledge, Perceptions, and Reactions towards the African Apefly (Spalgis lemolea lemolea) in Tanzania

This paper reports on farmers’ knowledge, perceptions, and reactions towards the African apefly (Spalgis lemolea lemolea), which appeared to be associated with some vegetables in some locations in Tanzania. Information was obtained from a sample of 100 key respondents using a semi-structured questionnaire and from focus groups selected from key locations in five districts in the country with histories with the African apefly. Acute and sub-acute toxicity tests of the African apefly were performed on female Swiss hybrid mice (Mus musculus) to assess whether or not the African apefly was toxic to mammals. The mice were exposed to increasing apefly meal concentrations in acute and sub-acute tests, and signs of toxicity were observed for 14 and 28 days, respectively. Blood samples were collected by cardiac puncture for hematological and biochemical analysis. Gross and microscopic examinations of the internal organs were done. The survey results showed that 92.1% of the respondents perceived the African apefly as poisonous and had stopped consuming the vegetables associated with it. In the toxicity tests, however, no death or toxic signs were displayed, and there was no significant difference between the control and treated mice in weight, hematological parameters, and histo-pathological examination results. These findings strongly indicate that, despite the negative perception by farmers regarding the African apefly, it is not poisonous. However, further studies on how farmers can be trained to have a positive perception of the African apefly and how the insect can be conserved for further research regarding its role in Tanzania are recommended.

Investigation of acute, subacute and subchronic toxicities of anthocyanin derived acylation reaction products and evaluation of their antioxidant activities in vitro.

Previously, anthocyanins were successfully acylated with lauric acid using Novozym 435 lipase, and the corresponding products were confirmed to have higher stability. As novel synthetic compounds, their toxicological safety has not been evaluated. Therefore, acute, subacute and subchronic toxicities of anthocyanin-lauric acid derivatives (ALDs) were investigated while their antioxidant activities were also evaluated in vitro. The acute toxicity results showed that the 50% lethal dose (LD50) of ALDs in mice was >10 g kg-1. Subsequently, the subacute toxicity test was conducted by oral administration of ALDs at doses of 0.63, 1.25 and 2.50 g kg-1 for 28 days. No adverse effect of ALDs on body weight, food/water intake, organ coefficient and histology was observed. Though there were some fluctuations in AST and ALT, the tested biochemical parameters were maintained within the normal ranges. The subchronic toxicity test results demonstrated that less than 0.60 g of ALDs per kg BW intake did not affect mortality, body weight, food/water intake, gross pathology, histology, hematology and serum biochemistry. Furthermore, cyanidin-3-(6''-dodecanoyl)-glucoside, the main component of ALDs, had a beneficial reducing power and a strong DPPH˙, ABTS+˙, and O2-˙ scavenging activity. This study provides an imperative reference to the safety of ALDs, suggesting their application as novel colorants or antioxidants in food and therapeutics.

Evaluation of Acute and Sub-acute Toxicity of the Aqueous Extract from the Fruit of Solanum indicum Linn. (Solanaceae) in Rats

Aim: The fruit of Solanum indicum Linn have been reported traditionally to have anti-hypertensive and diuretic properties. This study was undertaken to evaluate the toxicological potential of S. indicum fruits aqueous extract through the acute and sub-acute toxicity tests in rats.&#x0D; Methodology: For acute toxicity evaluation, a single oral dose of 5000 mg/kg of the plant extract was administrated in 60 days old female albino Wistar rats. Then, the animals were observed for 14 days. Sub-acute toxicity studies were conducted with 50 adult rats of both gender that orally received during 28 days, increasing doses of the plant extract. Their body weight and food intake were weekly collected. At the end of the experiment, biochemical and hematological parameters as well as histological analysis of organs (liver, kidneys and spleen) were undertaken.&#x0D; Results: Single oral administration of 5000 mg/kg dose of the fruit plant aqueous extract produced no mortality or signs of toxicity. During sub-acute test, no variations in body weight and food intake of both animals gender were observed. An important decrease in male’s rat liver weight were obtained at the dose 25 mg/kg; serum urea, total cholesterol, TAG, ALP and AST levels were significantly lowered in male especially at the dose 50 mg/kg, but this decrease was noticed only in serum urea, ALP and ALT in female rats. Furthermore, a significant decrease in platelets number, serum PCT, MPV and PDW levels were recorded in all treated male rats except those receiving the highest extract dose. No structural changes in treated animal organs section histology were observed when compared to controls.&#x0D; Conclusion: The fruits aqueous extracts of S. indicum is safe when administered acutely and for 28 days in rats. However, alterations on their hematological and biochemical parameters were not closely related with the dose, implying caution on its use.

Abstract 46: Three-Dimensional Scaffold-Free Spheroids with Fibroblast/Macrophage Co-Culture for in vitro Fibrosis Modeling

PURPOSE: An in vitro model of fibrosis is critical for designing surgical implants with better biocompatibility or new anti-fibrotic drug development. However, traditional 2D monolayer culture-based models of fibrosis show little resemblance to the in vivo fibrogenesis process. To enable screening of exponentially larger numbers of materials and conditions than can be achieved by sub-acute animal testing, we have developed a 3D scaffold-free spheroid system with human fibroblasts (spherical micro-tissue), which is more physiologically similar to real tissue. In addition, hybrid spheroids consisted of human fibroblasts and macrophages have been fabricated to investigate direct fibroblast-macrophage interaction and communication during fibrogenesis. METHODS: Spheroids were fabricated of 100% human fibroblasts with difference sizes (100, 200, 300,400, 500µm in diameter) respectively. Immunofluorescent staining with collagen-1 and apoptosis marker (Caspase-3) as well as qPCR with fibrosis genes (collagen-1 and aSMA) was performed to compare with the 2D control to identify the optimal size of the spheroids for an effective fibrosis model. In addition, we isolated human peripheral blood derived monocytes to differentiate into macrophages that could be used to populate the spheroids. Macrophages were co-cultured with fibroblasts under different ratios (1:2, 1:4, 1:8, and 1:16) to identify the ideal ratio in hybrid spheroids. We performed immunofluorescent staining with macrophages marker (MAC387) and fibroblasts marker (ER-TR7) and qPCR to optimize the ratio of macrophages in the hybrid spheroids to optimize similarity to physiologic in vivo fibrogenesis. RESULTS: Fibroblasts in spheroid had higher expression levels of fibrosis genes compared to the 2D monolayer control. The spheroid size of 200um showed the highest viability, more homogenous collagen deposition and the highest expression level of fibrosis genes due to a lower expression level of proteinase-related genes such as MMP1, MMP2 and MMP7. Interestingly, hybrid spheroids at a ratio of 1:16 (macrophage:fibroblast) showed higher survival rate of macrophages and a remarkably higher expression level of fibrosis genes (collagen-1, collagen-3 and TGFβ) with a lower expression level of anti-fibrotic MMP7. CONCLUSIONS: We have developed a 3D scaffold-free spheroid system as a more physiologically relevant fibrosis model in vitro for implant material and drug development and screening as compared to current methods. The inclusion of macrophages led to a significant change in fibroblast behavior. The size of the spheroids and the ratio of macrophages have been optimized regarding the resulting fibrosis profile. Future work will involve correlation of our findings with the in vivo tissue response.

Evaluation of the avian acute oral and sub-acute dietary toxicity test for pesticide registration

The United States Environmental Protection Agency (USEPA), as well as other international regulatory agencies, require pesticide registrants to submit toxicity data that are used to conduct ecological risk assessments. While the USEPA has required both an acute oral and sub-acute dietary test in birds, trends in the use of data from these tests over the past 20 years have suggested that the avian sub-acute dietary test generally does not contribute to risk assessment conclusions. To address this question, a retrospective analysis was conducted to evaluate 119 pesticides with publicly available ecological risk assessments that were registered into commerce between 1998 and 2017. New pesticides (i.e., registered in the United States within the past 20 years) were chosen for the retrospective analysis to show utility of these tests for modern pesticide chemistries. Risk quotient (RQ) values (a point estimate of exposure divided by a deterministic toxicity endpoint) from the avian acute oral and dietary tests, as well as risk assessment conclusions, were compared to determine which test(s) drove the risk assessment findings. The RQ values were chosen as the data point for comparison in order to assess total risk (i.e., exposure and toxicity). After comparing RQ values from avian acute oral versus sub-acute dietary tests, there was only one case in which an avian sub-acute dietary RQ was greater than the acute oral RQ. Thus, the sub-acute dietary test did not identify risk in greater than 99% (118 out of 119) of chemicals based on results that either the acute oral RQ was higher than the sub-acute dietary RQ, or both the acute oral and the subacute dietary tests did not generate an RQ value of concern. For the one exception, both the oral and sub-acute RQ values were greater than the USEPA's level of concern for endangered species. Based on the results of the retrospective analysis, it is concluded that in most cases avian risk can confidently be assessed without conducting the sub-acute dietary test.

Acute and 28-Day Repeated Inhalation Toxicity Study of Glycolic Acid in Male Sprague-Dawley Rats.

The use of glycolic acid is present in a variety of consumer products, including medicines, cleaners, cosmetics, and paint strippers. It has recently led to concerns about toxicity from inhalation exposure. Herein, the pulmonary toxicity of glycolic acid was investigated in rats. We conducted acute (~458 mg/m3) and sub-acute (~49.5 mg/m3) inhalation tests to identify the potential toxicities of glycolic acid. Inhalation exposure to glycolic acid in the acute and subacute inhalation tests did not cause any specific changes in clinical examinations, including body weight, organ weight, hematology, serum biochemistry, and histopathology. The polymorphonuclear neutrophils (PMNs) and inflammatory cytokines in Bronchoalveolar lavage fluid (BALF) increased in rats exposed to single and repeated inhalations. In the sub-acute test, the changes induced by glycolic acid were minor or returned to normal during the recovery period. The No Observed Adverse Effect Concentration (NOAEC) for the nasal and pulmonary toxicity of glycolic acid was determined to be over 50 mg/m3 at the end of a 28-day inhalation test in male rats.

Toxicological characterization and central nervous system effects of Calotropis procera Ait. aqueous extracts in mice

Objective: To evaluate the toxicological and psychotropic properties of Calotropis (C.) procera. Methods: C. procera leaves and root-bark aqueous extracts were evaluated for their toxic and behavioral effects using adult mice. Toxicity studies were carried out using Organisation for Economic Cooperation and Development guidelines 423 and 407 for acute and subacute evaluation. Behavioral studies were performed using traction test, fireplace test, hole-board test and forced-swimming test to evaluate the sedative, anxiety and depressive-like activities of the extracts. Results: Very low acute toxicity was observed in mice that received both leaves and root-bark extracts. The subacute test showed some morphological, biochemical and hematological changes in the treated groups. Behavioral assessment demonstrated anxiety effects on mice for C. procera leaf extract (400 mg/kg of body weight). Conclusions: The acute use of C. procera (leaves and root-barks) aqueous extracts could be considered as low toxic. However, their repeated uses could have harmful effect on some organs. Likewise, a single dose up to 400 mg/kg body weight of these extracts produce no sedative or depressive-like effect, but they possess possible dose dependent anxiety effect. Yet, more studies are necessary to relate these results to the chemical profile of the plant extracts.

Preclinical testing of 5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide: a potent protein kinase C-ι inhibitor as a potential prostate carcinoma therapeutic.

Protein kinase C-iota (PKC-ι) is an oncogene overexpressed in many cancer cells including prostate, breast, ovarian, melanoma, and glioma. Previous in-vitro studies have shown that 5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide (ICA-1s), a PKC-ι specific inhibitor, is effective against some cancer cell lines by decreasing cell growth and inducing apoptosis. To assess ICA-1s as a possible therapeutic, in-vivo studies using a murine model were performed. ICA-1s was tested for stability in blood serum and results demonstrated that ICA-1s was stable in human plasma at 25 and 37°C over a course of 2 h. Toxicity of ICA-1s was tested for both acute and subacute exposure. The acute exposure showed patient surviving after 48 h of doses ranging from 5 to 5000 mg/kg. Subacute tests exposed the patients to 14 days of treatment and were followed by serum and tissue collection. Aspartate aminotransferase, alkaline phosphatase, γ-glutamyl transpeptidase, troponin, and C-reactive protein serum levels were measured to assess organ function. ICA-1s in plasma serum was measured over the course of 24 h for both oral and intravenous treatments. Heart, liver, kidney, and brain tissues were analyzed for accumulation of ICA-1s. Finally, athymic nude mice were xenografted with DU-145 prostate cancer cells. After tumors reached ~0.2 cm2, they were either treated with ICA-1s or left as control and measured for 30 days or until the tumor reached 2 cm2. Results showed tumors in treated mice grew at almost half the rate as untreated tumors, showing a significant reduction in growth. In conclusion, ICA-1s is stable, shows low toxicity, and is a potential therapeutic for prostate carcinoma tumors.

Determination of mancozeb toxicity and biochemical effects in common carp (Cyprinus carpio)

The aim of this study was to investigate mancozeb toxicity and its effects on physiological characteristics of common carp. Fish were reared for one week to acclimatize with the experimental conditions. For 96h-LC50 determination, the fish were stocked in 16 aquaria at the density of 10 fish per aquarium. The aquaria were exposed to 8 mancozeb concentrations (two aquaria per concentration) for 96 h (0, 0.94, 1.87, 3.75, 7.50, 15, 30 and 60 mg/L). 96h-LC50 was calculated based on the fish mortality, being 8.03 (4.95-13.2) mg/L. For sub-acute test, the fish were exposed to 0 (control), 1.6, 2.4 and 3.2 mg/L mancozeb (20, 30 and 40% of the 96h-LC50) for one week. Blood samples were taken from each treatment for determination of plasma glucose, total protein, albumin, globulin, calcium, alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Mancozeb exposure led to increase in glucose and AST, and decrease in plasma proteins and ALT. In conclusion, mancozeb exposure causes stress response, health problem and tissue damage in common carp.

Acute and Subacute Toxicity Evaluation of the Stem Bark Aqueous Extract of Harungana Madagascariensis in Rodents

The present investigation was carried out to evaluate the safety of a stem bark aqueous extract of Harunganamadagascariensis Lam. (Hypericaceae) by determining its potential toxicity after acute and subacute administration in rodents. Acute toxicity tests were carried out in mice and the behavior, death and median lethal dose (LD50) were estimated. Subacute toxicity (28 days) studies were conducted in rats with oral daily doses of 200, 400 and 600 mg/kg. Parameters observed at the end of the subacute tests included changes in body and vital organ weights, mortality, hematological, biochemical, hepatic and kidney effects. Harunganamadagascariensisextract did not produce any visible toxicity or mortality with oral doses up to 2000 mg/kg within 14 days of single treatment, leading to the conclusion that the LD50 is greater than 2000 mg/kg. In the subacute toxicity tests, neither mortality nor visible signs of lethality was seen in rats. No significant change in the weight of the kidney, liver, heart, lungs spleen, pancreas and testicles was observed. Alanine transaminase (ALT) increased significantly in males at 400 and 600 mg/kg, whereas Aspartate transaminase (AST) decreased at 600 mg/kg in female rats. HDL Cholesterol was reduced at 600 mg/kg in female rats. There was a significant increase in urea concentration in female rats at 400 mg/kg. A significant decrease, both in platelet volume distribution (PVD) at 400 mg/kg in male rats and in red cell volume distribution (RDW) at 200 mg/kg were recorded in female rats respectively, but with no changes in other hematologic parameters. Histological study shows normal structure of liver, kidneys and heart of control and treated rats. Results indicate that oral doses of aqueous stem bark of Harunganamadagascariensis are relatively safe in rats; however, assessment of hepatobiliary function should be done during chronic use in humans.

Antidepressant-like action of agmatine in the acute and sub-acute mouse models of depression: a receptor mechanism study

Previous studies have shown that agmatine, a potential neuromodulator or co-transmitter, exhibited antidepressant-like action in animal models, yet its mechanism, especially the receptor mechanism, remains unclear. In the present study, using efaroxan, a preferential antagonist of I1 imidazoline receptor (I1R) and yohimbine, an antagonist of α2 adrenergic receptor (α2AR), we investigated the roles of I1R and α2AR in agmatine's antidepressant-like effect in acute and sub-acute depression models in mice. We found that in the tail-suspension test (TST) and the forced swimming test (FST), acute administration of agmatine (20 and 40mg/kg, p.o.) significantly shortened the immobility time. Concurrent administration of efaroxan (1mg/kg, i.p.) completely abolished the antidepressant-like effects of agmatine (40mg/kg, p.o.) whereas yohimbine (5mg/kg, i.p.) failed to exert similar effects, suggesting that the acute antidepressant-like effects of agmatine was mainly mediated by I1R but not α2AR. Additionally, in the learned helplessness (LH) test, repeated administration of agmatine (20mg/kg, p.o., q.d.) for 5days significantly decreased the escape latency and the number of escape failure, and these effects were respectively abolished by concurrent administration of efaroxan (0.5mg/kg,i.p., q.d.) and yohimbine (3mg/kg, i.p., q.d.) for 5days, suggesting that the antidepressant-like action of agmatine in the LH test was achieved via the activation of both I1R and α2AR. In summary, we found that the antidepressant-like effects of agmatine in the TST and the FST were mediated by activating I1R and in the sub-acute LH test were mediated by activating both I1R and α2AR.

Abstract 5868: Preclinical testing of 5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide (ICA-1): A potent protein kinase C-ι inhibitor as a potential anticancer drug

Abstract Protein Kinase C iota (PKC-ι) is an oncogene overexpressed in many cancer cells including prostate, breast, ovarian, melanoma and glioma. Previous In- vitro studies have shown that ICA-1 a PKC-ι specific inhibitor, is effective against some cancer cell lines by decreasing cell growth and inducing apoptosis {Pillai et al. Int. J. Biochem. Cell Biol. 43 (2011) 784-794. To assess ICA-1 as a possible therapeutic, In- vivo studies using a murine model were performed. ICA-1 was tested for stability in blood serum and results demonstrated that ICA-1 was stable in human plasma at 25oC and 37oC over a course of two hours. Toxicity of ICA-1 was tested for both acute and sub-acute exposure. The acute exposure showed subject surviving after 48 hours of doses ranging from 5mg/kg to 5000 mg/kg. Sub-acute tests exposed the subjects to 14 days of treatment and was followed by serum and tissue collection. AST, ALK-P, GGT, Troponin, and CR-P serum levels were measured to assess organ function. Heart, liver, kidney, and brain tissues were analyzed for accumulation of ICA-1. Finally athymic nude mice were xenografted with DU-145 prostate cancer cells. After tumors reached approximately 1 cm2, they were either treated with ICA-1 or left as control until the tumor reached 2 cm2. Results showed tumors in treated mice took almost double the time to reach the experimental end point, showing a significant growth reduction. In conclusion, ICA-1 is stable, shows low toxicity, and is a potential therapeutic for some carcinoma tumors. Citation Format: Christopher Apostolatos, Andre H. Apostolatos, Wishrawana S. Ratnayake, Marie Bourgeois, Mildred Acevedo-Duncan. Preclinical testing of 5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide (ICA-1): A potent protein kinase C-ι inhibitor as a potential anticancer drug [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5868.