Oral Administration Of High Doses Research Articles

Comparative Study on the Effect of Bacopa monniera and Omega3 Fatty Acids on the Cortisol Hormone Level in Cold Stress Induced Wistar Albino Rats

In recent years, researchers have identified several natural compounds that could potentially help to retard mental stress. Extracts of Bacopa monniera (BM), a traditional ayurvedic medicine and omega3 faffy acids (OFA) have been reported to enhance effects mental functions by combating the stress. However, there is still an lacunae studies on which of those two drugs has better results on cortisol hormonal changes due to stress. The current study aimed to compare the effects of BM and omega3 fatty acids on cold stress induced Wistar rats. We divided the animals into seven groups. Group I was control in which rats were kept under ideal laboratory conditions, Group II was given cold water swim stress for a period of one month after which the rats were sacrificed for hormonal assay, Group III in which cold water swim stress given for a month followed by oral administration of normal saline as vehicle for a month, Group IV and V in which cold water swim stress given for a month followed by oral administration of BM extracts 40mg/kg and 80mg/kg respectively as treatment for a month. Group VI and VII includes cold stress followed by oral gavages of OFA 60mg/Kg of Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) for 2 weeks (short-term model) and 4 weeks (long term model) respectively. After the treatment period blood was collected and cortisol assay was done. The data were compared with those of control rats. The results showed an increase in the cortisol level in the stressed group of rats when compared to the treatment group. These results indicate that oral administration of high dose of BM and long-term administration of OFA improves the cortisol level and helps in controlling the stress in rats.

Prospects of Intravenous Coenzyme Q10 Administration in Emergency Ischemic Conditions.

Coenzyme CoQ10 (CoQ10) is an endogenous lipid-soluble antioxidant that effectively protects lipids, proteins, and DNA from oxidation due to its ability to undergo redox transitions between oxidized and reduced forms. Various oxidative stress-associated infectious and somatic diseases have been observed to disrupt the balance of CoQ10 concentration in tissues. As a high molecular weight polar lipophilic compound, CoQ10 exhibits very limited oral bioavailability, which restrains its therapeutic potential. Nevertheless, numerous studies have confirmed the clinical efficacy of CoQ10 therapy through oral administration of high doses over extended time periods. Experimental studies have demonstrated that in emergency situations, intravenous administration of both oxidized and reduced-form CoQ10 leads to a rapid increase in its concentration in organ tissues, offering protection for organ tissues in ischemic conditions. This suggests that the cardio- and neuroprotective efficacy of intravenously administered CoQ10 forms could present new opportunities in treating acute ischemic conditions. Based on these findings, the review provides reasoning supporting further research and implementation of CoQ10 dosage forms for intravenous administration in emergency situations into clinical practice.

Preparation of pazopanib-fumarate disodium glycyrrhizinate nanocrystalline micelles by liquid-assisted ball milling

Pazopanib (PZ) is a multikinase inhibitor, which is mainly used in the treatment of soft tissue sarcoma and advanced renal cancer. However, because of its water insolubility, oral bioavailability is poor. At the same time, photo lability and high dose oral administration lead to severe hepatotoxicity, which is limited in clinical application. In this paper, the novel pazopanib-fumarate disodium glycyrrhizinate nanocrystalline micelles are successfully prepared by liquid-assisted ball milling. The prepared cocrystals and nanocrystalline micelle structures are systematically characterized by X-ray diffraction (XRD), differential scanning calorimetry (DSC) and Fourier Transform Infrared Spectrometer (FTIR) analysis. In vitro solubility and dissolution experiments show that the solubility and dissolution of nanocrystalline micelles are significantly improved under different simulated physiological conditions. The accelerated stabilization experiments show that the nanocrystalline micelles have good physical and chemical stability and showed excellent stability in water (Zeta potential was 62.39 mV). In addition, the in vivo bioavailability of nanocrystalline micelles is 3 times higher than that of PZ, and the therapeutic threshold (> 20 μg/mL) is up to 30 h. This new strategy provides a feasible solution to the undesirable properties of PZ.

Toxicity of Toddalia asiatica is associated with microcephaly and hypochondroplasia in mice

In Kenya various plant extracts are widely used as folklore remedies for various ailments including malaria etc. However, use of these traditional remedies poses a risk to the users due to the scarcity of data on their safety including their teratogenic potential. Hence this study evaluated the teratogenic and sub-acute toxic effects of methanol: dichloromethane extract of Toddalia asiatica in animal mice. In the study, young female mice aged between 6-7weeks were mated with mature males and conception confirmed using vaginal plug, maternal weight and breast enlargement. Administration of the dosed of the plant extract and controls was done from day 6 through 15 of gestation. The mice were euthanized on the 19th day of gestation. The weight of the gravid uterus, pups, number of pups and gross examination to identify any physical abnormalities was done. In the toxicity study male mice aged between 6-7 weeks received oral administration of high doses of plant extract and the control daily for 28days. On the 29th day animals were sacrificed, cardiac puncture conducted and collected blood for hematology and biochemistry analysis. The data obtained was analyzed using one way ANOVA followed by Tukey as the post hoc test. Statistical significance was set p < 0.05. The extracts caused significant reduction (p<0.001) in head size, birth weights and length of the limbs but no effects on hematology parameters. Hence the plant extract exhibited teratogenic effects and thus should be used with care during pregnancy.

Salicylic acid-based hypoxia-responsive chemodynamic nanomedicines boost antitumor immunotherapy by modulating immunosuppressive tumor microenvironment

The delivery of salicylic acid or its derivatives to tumor tissue in the form of nanomedicine is critical for the studies on their potential synergistic mechanism in tumor therapy and chemoprevention considering the dangerous bleeding in the high-dose oral administration. To deepen the understanding of their role in adjusting immunosuppressive tumor microenvironment (ITM), herein, we firstly developed a hypoxia-sensitive Fe-5,5′-azosalicylic acid nanoscale coordination polymer nanomedicines (FeNCPs) via a “old drugs new tricks” strategy for synergistic chemodynamic therapy (CDT) and remodulation of ITM to elevate antitumor immunotherapy effect. PEGylated FeNCPs could be reductively cleaved to release 5-aminosalicylic acid (5-ASA) and ferric ions by azo-reductase under hypoxic conditions, which could induce tumor cell death by Fenton reaction-catalysis enhanced CDT and 5-ASA-converted carboxylquinone to promote the production of •OH. Meanwhile, cyclooxygenase-2 (COX-2) and its enzymatic product prostaglandin E2 (PGE2), as immune negative regulatory molecules, can promote tumor progression and immune tolerance. The released 5-ASA as a COX inhibitor could suppress the expression of PGE2, and Fe3+ was employed to reeducate M2-like tumor-associated macrophages (TAMs) to M1-like phenotype, which could initiate antitumor immune response to reach better antitumor immunotherapy. This work broadens the application of salicylic acid derivatives in antitumor immunotherapy, and provides a new strategy for their “old drugs new tricks”. Statement of SignificanceCyclooxygenase-2 (COX-2) and its enzymatic product prostaglandin E2 (PGE2), as immune negative regulatory molecules, facilitate the differentiation of immune cells into immunosuppressive cells to build the immunosuppressive tumor microenvironment, which can promote tumor progression and immune tolerance. Thus, down-regulation of COX-2/PGE2 expression may be a key approach to tumor treatments. Meanwhile, as a class of inhibitors of COX-2/PGE2, the potential mechanism of aspirin or 5-aminosalicylic acid has been a mystery in tumor therapy and chemoprevention. To expand the application of aspirin family nanomedicine in biomedicine, herein, we firstly developed a hypoxia-sensitive Fe-5,5′-azosalicylic acid nanoscale coordination polymer nanomedicines via a “old drugs new tricks” strategy for synergistic chemodynamic therapy and remodulation of immunosuppressive tumor microenvironment to elevate antitumor immunotherapy effect.

Oral administration of Lactiplantibacillus plantarum 22A-3 exerts anti-allergic activity against intestinal food allergy mouse models sensitized and challenged with ovalbumin

In recent years, researches on food components with anti-allergic effects have been gathering attention, because of the expectation for the establishment of a safe and effective treatment for food allergy. Previous studies have reported that Lactiplantibacillus plantarum 22A-3 (LP22A3) inhibited degranulation of mast cells and reduced IgE production. We have developed a gastrointestinal allergy system in which mice are sensitized by intraperitoneal and oral administration of OVA and then challenged by oral administration of high doses of OVA. As a result, an increase in the amount of IgE in the blood and a decrease in the temperature of the colon were confirmed, and it was clarified that food allergy was induced by oral administration of high dose of OVA as the challenge. Oral administration of LP22A3 ameliorated allergic responses significantly by reducing the amount of IgE in the blood and to recovered the decrease of rectal temperature. However, LP22A3 did not affect the intestinal barrier function. Administered LP22A3 significantly suppressed mRNA expression of OX40L and IL-4. These results suggested that LP22A3 suppressed Th2 differentiation and IL-4 production via downregulation of OX40L, and consequently suppressed IgE production. LP22A3 might provide a safe and effective treatment for allergic diseases due to ability modulating intestinal immune system.

CU06-1004 modulates the adenosine monophosphate (AMP)-associated protein kinase (AMPK) signaling pathway and inhibits lipogenesis in 3T3-L1 adipocytes and high-fat diet-induced obese mice

AimsThe endothelial dysfunction blocker CU06-1004 exhibits anti-inflammatory effects in chronic diseases. Obesity is a major cause of chronic inflammation, and the effect of CU06-1004 on obesity has not been studied yet. Therefore, in this study, we investigated the anti-obesity properties of CU06-1004 in 3T3-L1 adipocytes and high-fat diet-induced obese mice. MethodsDifferentiated 3T3-L1 adipocytes were treated with various concentrations of CU06-1004 (0–20 μg/mL) and subjected to Oil Red O staining to determine the levels of lipid droplet and intracellular triglyceride accumulation. Additionally, high-fat diet-induced obese C57BL/6J mice were administered with a low (10 mg/kg/day) or high (20 mg/kg/day) oral dose of CU06-1004. Finally, the expressions of genes and proteins involved in the adenosine monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway were assessed by real-time polymerase chain reaction and Western blot, respectively. Key findingsThe CU06-1004 administration reduced lipid accumulation in the 3T3-L1 adipocytes by inhibiting the expressions of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein alpha, fatty acid binding protein 4, and fatty acid synthase in a dose-dependent manner. Additionally, it significantly increased the phosphorylation of AMPKα and acetyl-CoA carboxylase in the 3T3-L1 adipocytes. An oral administration of high dose of CU06-1004 in the obese mice significantly decreased their body weight and the mesenteric white adipose tissue weight. Furthermore, CU06-1004 improved hepatic steatosis by reducing lipogenesis, besides improving insulin resistance and exerting systemic anti-inflammatory effects. SignificanceCU06-1004 may have therapeutic potential in the prevention of obesity and obesity-related disorders.

Abstract P243: The IRAK4 inhibitor CA-4948 demonstrates antitumor activity in a preclinical model of CNS lymphoma

Abstract Background: Central nervous system (CNS) lymphoma, both primary and secondary disease, carries the worst prognosis of all Non-Hodgkin’s lymphoma with a 5 year survival of less than 30% with current therapeutic options. Recent studies have shown constitutive activation of both NF-κB and MAPK signaling pathways in the maintenance and progression of CNS lymphoma. This often coincides with mutations in MYD88. IRAK4 is a serine/threonine kinase that mediates interleukin 1 receptor (IL-1R) and Toll-like receptor (TLR) signaling downstream of MYD88, and is a novel therapeutic target for this disease. CA-4948 is an oral first-in-class small molecule inhibitor of IRAK4 that has demonstrated clinical activity in patients with systemic Non-Hodgkin’s Lymphoma. Our study goal was to evaluate CA-4948 efficacy in a syngeneic preclinical model of primary CNS B cell lymphoma (A20), supporting future clinical investigation of this agent in this difficult to treat disease. Materials and Methods: Plasma, cerebrospinal fluid (CSF), and brain tissue (both naïve and tumor bearing) were assessed for CA-4948 drug concentration following single high-dose oral administration using UPLC-MS/MS. Biomarker expression, including NF-κB and MAPK, was measured in control and CA-4948 treated brain tissue resected from tumor-bearing animals using multi-parameter immunohistochemistry and 3D modeling of cleared tissue. Dose-dependent survival responses were also measured using the A20 preclinical model of primary CNS lymphoma. Results: While brain concentrations of CA-4948 remained low with respect to plasma (~4-5%), therapeutic dose levels were attained in both CSF and brain tissue (>502 nM). No significant difference between naïve and A20 tumor bearing brain tissue was observed, indicating the blood brain barrier remained intact in this model. Biomarker analyses revealed significant downregulation of both MAPK and NF-κB signaling pathways in response to CA-4948, as evidenced by reduced protein levels of phospho-p38Thr180/Tyr182, phospho-ERK1/2Thr202/Tyr204, and phospho-p65Ser536 in both the tumor cells and the tumor microenvironment. Finally, CA-4948 treatment produced a dose-dependent survival response in syngeneic animals harboring CNS A20 tumors. Conclusions: CA-4948 is able to reach therapeutic dose levels in the CNS in a preclinical murine model of CNS lymphoma, producing significant and dose-dependent anti-tumor activity and survival advantage. These data support further evaluation of CA-4948 in clinical investigation and treatment of patients with CNS lymphoma. Citation Format: Christina A. von Roemeling, Bently P. Doonan, Lan Hoang-Minh, Han W. Tun, Elizabeth Martinez, Raul Soikes, Reinhard von Roemeling, Duane A. Mitchell. The IRAK4 inhibitor CA-4948 demonstrates antitumor activity in a preclinical model of CNS lymphoma [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P243.

Hepato-renal toxicity of Myristica fragrans Houtt. (Myristicaceae) seed extracts in rats

ObjectivesMyristica fragrans is used in many parts of the world as a common spice and herbal medicine for managing a wide variety of ailments. The rich nutritional factors in M. fragrans seeds show potential application in food biofortification; however, the long-term safety of using a high dose of the seeds needs to be validated. This study seeks to investigate how oral administration of high doses of methanol (ME) and n-hexane (NHE) extracts of M. fragrans seed for one or two weeks affects the histology and serum markers of kidney and liver of rats. MethodsAdult male Swiss mice (6–7 weeks, 20–30 g) were used for acute toxicity study by standard methods, while male Wistar rats (6–8 weeks, 100–120 g) were used for sub-acute toxicity test. The sixty rats were distributed into five groups of 12 rats each: Group 1 received normal saline, groups 2 and 3 were orally treated with 500 and 1000 mg/kg b.w/day ME while groups 4 and 5 were fed 500 and 1000 mg/kg b.w/day NHE, respectively for 14 days. Six rats from each group were sacrificed on days 8 and 15 after 24 h of fasting. Markers of kidney and liver status of test and control animals were compared using one-way analysis of variance. ResultsThe presence of terpenoids, flavonoids, alkaloids, phenols, steroids, and tannins were detected in both extracts, although at varying levels. There was no obvious sign of toxicity nor mortality in acute toxicity test after 24 h of administration of extracts up to 5000 mg/kg b.w. However, there were significant (p < 0.5) elevations in urea, total bilirubin and creatinine concentrations, alkaline phosphatase, aspartate and alanine aminotransferases, and lactate dehydrogenase activities in rats fed extracts for seven or fourteen days relative to control. Furthermore, the n-hexane extract at 1000 mg/kg elicited some histological changes consistent with hepatotoxicity. ConclusionsAlthough the extracts were rich in some essential phytochemicals, this study demonstrated that long-term administration of high doses of the extracts elicits hepato-renal toxicities. Hence, consuming a large amount of the seed over a long duration is discouraged.

Nicotinamide Improves Anemia and Decreases Reactive Oxygen Species in Sickle Cell Disease Mice

The polymerization of deoxygenated HbS molecules in the red blood cells (RBCs) of patients with sickle cell disease (SCD) causes RBC destruction resulting in chronic pain, debilitating acute pain crises, strokes, multi-organ damage and a reduced life span. Therapeutic options remain limited. Bone marrow transplantation can be curative but is not an option for the majority of patients. Gene therapy interventions that also offer the promise of a cure are under investigation but are not likely to be available to the vast majority of patients in the near future. Hydroxyurea, the first drug approved for treatment of SCD, increases levels of Fetal Hemoglobin (HbF) that inhibit polymerization of HbS molecules but is not effective in all patients while a more powerful HbF-inducing drug, the DNA methyltransferase inhibitor decitabine has yet to be approved. L-glutamine, another approved therapeutic option, increases NAD redox potential and decreases reactive oxygen species (ROS) in the sickle RBCs to reduce symptoms. In our laboratory we have observed that increased ROS is associated with the retention of mitochondria in the SCD RBCs and have hypothesized that the abnormal presence of mitochondria in these cells is a major source of ROS (Jagadeeswaran et al Exp Hematol 50:46-52, 2017). In this investigation we have tested the hypothesis that chronic oral supplementation with nicotinamide, a direct precursor of NAD synthesis, would improve NAD redox potential, decrease mitochondrial retention and ROS in SCD RBCs, and reduce anemia in the SCD mouse model. The effect of nicotinamide was tested in SCD mice whose drinking water was supplemented for three months with 1% nicotinamide. The percentage of RBCs retaining mitochondria and the levels of ROS were determined by flow cytometric assays using the mitochondrial-specific dye TMRM and the ROS probe CM-H2DCFDA, respectively. In SCD mice receiving nicotinamide the fraction of RBCs retaining mitochondria was reduced 22.1% (p<0.05) and the level of ROS in RBCs was reduced 41% (p<0.01) compared to control SCD mice. The reticulocyte percentage was reduced 28% in nicotinamide-treated SCD mice compared to control SCD mice (p<0.01). The total RBC count was 30% higher (p<0.05) in nicotinamide-treated mice (6.61±0.76 X 10 6/μl) compared to control SCD mice (5.08±0.70 X 10 6/μl). Similar differences in hematocrit and total hemoglobin were also observed but failed to reach statistical significance. Total NAD levels were not significantly different in SCD mice receiving nicotinamide compared to control SCD mice (p<0.05), but the NADH/NAD total ratio was increased 2 fold (p<0.05). These results show that oral administration of high doses of nicotinamide decreases mitochondrial retention and ROS in SCD RBCs and improves NAD redox potential and anemia in SCD mice. These effects strongly suggest that additional studies be performed to investigate nicotinamide as a therapeutic option in SCD. [Display omitted] DisclosuresSaunthararajah: EpiDestiny: Consultancy, Current holder of individual stocks in a privately-held company, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.

Improving Reproductive Performance and Health of Mammals Using Honeybee Products.

Honeybee products have positive effects on the reproductive performance of mammals. Many honeybee product constituents are biologically active, with antioxidant, antimicrobial, antiviral, anti-inflammatory, immunomodulatory, antifungal, wound-healing, and cardio-protective properties. Honeybee products also improve male and female fertility rates by enhancing gamete cryopreservation, in vitro maturation and fertilization, and embryo development. Previously published studies confirmed their efficacy for alleviating reproductive toxicity caused by contaminants and lifestyle habits that impair overall health and well-being. However, high-dose oral administration of honeybee products may adversely affect the reproductive system, and unfavorable effects were alleviated by treatment cessation. For this reason, this review proposes that bioactive components from bee products can be used as a strategy for improving the reproductive performance and health of mammals.

Safety of oral administration of high doses of ivermectin by means of biocompatible polyelectrolytes formulation

The FDA-approved drug ivermectin is applied for treatments of onchocerciasis and lymphatic filariasis. The anti-cancer and anti-viral activities have been demonstrated stressing possibilities for the drug repurposing and therefore new information on high dosage safety is on demand. We analyzed in vivo tissue responses for high doses of ivermectin using Corydoras fish as animal model. We made intestinal histology and hematologic assays after oral administration of ivermectin transported with polyelectrolytes formulation. Histology showed any apparent damage of intestinal tissues at 0.22–170 mg of ivermectin/kg body weight. Immunofluorescence evidenced delocalization of Myosin-Vb at enterocytes only for the higher dose. Hematology parameters showed random variations after 7 days from administration, but a later apparent recover after 14 and 21 days. The study evaluated the potential of high doses of oral administration of ivermectin formulation, which could be an alternative with benefits in high compliance therapies.

Selective laser sintering (SLS) technique for pharmaceutical applications—Development of high dose controlled release printlets

The paper presents the unique approach to 3D printing by Selective Laser Sintering of high dose controlled release pharmaceutical dosage form, which contains almost exclusively drug substance - just crystalline paracetamol as a model drug substance and small amount of dye were used. Comprehensive printlet characterization, that included pore space analysis, drug release and subsequent dissolution modeling as well as exploratory analysis, revealed various degrees of freedom for tuning its functional properties i.e. dissolution performance. Two degrees of freedom were found substantial for the proposed approach: (1) the macro structure shaping in terms of active surface to volume ratio; (2) microstructure shaping in terms of pore space structure. The latter was found to have a great potential and can be modified by the manufacturing parameters, e.g. hatch spacing. The simplicity of the preparation and flexibility to control the dissolution performance of the pharmaceutical dosage are the advantages of the proposed approach. • Pharmaceutical high dose oral administration. • Highly drug-loaded controlled release formulations by selective laser sintering • Macro and micro structure shaping for drug dissolution tuning. • Flexible dosage form design for personalized medicine. • Simple manufacturing and simple dissolution mechanism.

(Solvato-) polymorphism of formulations of rifampicin for pulmonary drug delivery prepared using a crystallization/spray drying process.

Rifampicin is an antibiotic used in tuberculosis therapy showing extensive (solvato-) polymorphism. Per oral administration of high doses is recommended, but application as dry powder for inhalation at the site of infection being the lungs, is desirable. Recrystallization from ethanol and consecutive spray drying is reported to yield a rifampicin dihydrate with suitable aerosol performance and stability. Nevertheless, the origin of water in the crystal remained unclear and demanded further investigation so to clarify its solid state throughout manufacture and storage. The present study reports the relationship of (solvato-) polymorphs occurring during manufacture and storage of samples recrystallized and spray dried from ethanol, methanol and water and it was concluded that processes involving a recrystallization from EtOH and MeOH produce particles of a common isostructural group of channel solvates. Samples recrystallized and spray dried from water were identified as members of another isostructural group, which was already characterized in literature. As a second aim, aerosol performance and storage stability of the formulations were investigated and all samples showed stable aerosol performance. Chemical stability of samples spray dried from ethanol was found suitable over a period of six months, whereas samples spray dried from methanol or water showed significant degradation.

Inhalable formulations of rifampicin by spray drying of supersaturated aqueous solutions

Tuberculosis is still one of the leading causes of death from a single infectious agent (i.e. Mycobacterium tuberculosis). First line therapy includes per oral administration of high doses of rifampicin over several months and is often times accompanied by the occurrence of unwanted side effects that might limit the patient’s adherence to the therapy. Thus, local antibiotic treatment at the site of infection i.e. the lungs is desirable. Amongst other approaches, spray drying of solutions of rifampicin has been shown as suitable method to produce respirable dry powders. In this work, we present inhalable formulations manufactured via spray drying of aqueous solutions of rifampicin. Powders manufactured were characterized for their aerodynamic and solid state properties, as well as their physical and chemical stability. The main focus of this study was to investigate the mechanism of particle formation using an acoustic levitator. Fine particle fractions of the test formulations ranged from 80 to 89% whereas a reference formulation (a spray dried isopropyl alcoholic solution of rifampicin) showed a lower fine particle fraction of 37%. Acoustic levitator and surface tension experiments showed that interfacial properties of rifampicin lead to early crust formation upon drying of the droplets, which eventually decoupled from the liquid core and formed highly collapsed, low apparent density powders with excellent aerosol properties.

Grape-Leaf Extract Attenuates Alcohol-Induced Liver Injury via Interference with NF-κB Signaling Pathway.

Grape (Vitis vinifera) leaf extracts (GLEs) are known to be rich in phenolic compounds that exert potent antioxidant effects. Given the vulnerability of the liver to oxidative damage, antioxidants have been proposed as therapeutic agents and coadjuvant drugs to ameliorate liver pathologies. The current study was designed to characterize secondary metabolites and investigate the hepatoprotective effects of GLE and its underlying mechanisms. The secondary metabolites were profiled using HPLC–PDA–ESI-MS, and forty-five compounds were tentatively identified. In experimental in vivo design, liver injury was induced by oral administration of high doses of ethanol (EtOH) for 12 days to male Sprague Dawley rats that were split into five different groups. Blood samples and livers were then collected, and used for various biochemical, immunohistochemical, and histopathological analyses. Results showed that GLE-attenuated liver injury and promoted marked hepatic antioxidant effects, in addition to suppressing the increased heat-shock protein-70 expression. Moreover, GLE suppressed EtOH-induced expression of nuclear factor-κB (NF-κB) p65 subunit and proinflammatory cytokine tumor necrosis factor-α. Caspase-3 and survivin were enhanced by EtOH intake and suppressed by GLE intake. Finally, EtOH-induced histopathological changes in liver sections were markedly normalized by GLE. In conclusion, our results suggested that GLE interferes with NF-κB signaling and induces antioxidant effects, which both play a role in attenuating apoptosis and associated liver injury in a model of EtOH-induced liver damage in rats.

Toxicological research and safety consideration of coumarins

Coumarin is an important class of natural organic compounds, which widely exists in a variety of plants and microorganisms. Coumarins have many biological activities and wide clinical applications, such as anti-tumor, anti-HIV, anti-bacterial, anti-inflammatory, anti-oxidation, anti-coagulation, but they have obvious toxic effects in rodents. It was found that the toxicity of coumarins in different animals and organs was significantly different, and high dose oral administration was more likely to produce toxic reactions. Based on the research and analysis of domestic and foreign literatures in recent 60 years, this paper mainly summarized the hepatotoxicity and pulmonary toxicity induced by coumarins, and probed into their possible mechanisms. It was found that the toxicity of coumarins had metabolic differences and species differences. The liver of rats and lungs of mice were more susceptible to coumarins. Toxic reactions occurred mainly in the second metabolic pathway of coumarin metabolism in vivo. In order to put forward safety considerations and evaluate the impact of coumarin on human body, it was found that coumarin is unlikely to produce hepatotoxicity at normal exposure level. It was also suggested that species differences due to different metabolic patterns in model animals should be carefully considered when assessing coumarin toxicity, in order to provide reference for clinical research and rational use of coumarins and improve the rational use of coumarins.

Infectious complications as predictors of adverse outcome in a patient with systemic lupus erythematosus (clinical case)

Systemic lupus erythematosus is a systemic autoimmune disease. Over the past decades, great success has been achieved in its treatment. However, mortality in systemic lupus erythematosus is still high. An adverse outcome may occur because of kidney damage, damage of the nervous system, severe hematological disorders, etc. The factor of unfavorable prognosis is infectious complications. The article presents a prospective clinical observation of a faytal case of severe course of systemic lupus erythematosus. The patient was treated in the rheumatology department of the State Regional Clinical Hospital in Saratov from 2007 to 2014. The presence of high disease activity, multiple system disorders — polyarthritis, lupus nephritis, hepatitis, leukopenia, and recurrent necrotizing cutaneous vasculitis required the administration of high doses of immunosuppressive drugs. The progressive course of the disease, resistance to hormonal therapy, the rapid development of infectious complications made the prognosis for the patient’s life extremely unfavorable. Persistent autoimmune leukopenia was the background condition for the development of complications. We used combined therapy — the oral administration of high doses of glucocorticoids together with intravenous injections (pulse therapy), broad-spectrum antibacterial drugs, intravenous immunoglobulin, drugs that improve tissue trophy and microcirculation. It was possible to decrease the disease activity, and also to restore the functional activity of the patient, to maintain low disease activity for 3 years, to prolong the life of a young patient by 8 years. The adverse combination of high activity of systemic lupus erythematosus with recurrent soft tissue infections caused difficulties for the therapy. Administration of adequate doses of cytostatic and biological agents were impossible due to leukopenia and secondary infection, which led to the death of the patient.

Resistance of parvalbumin to gastrointestinal digestion is required for profound and long-lasting prophylactic oral tolerance.

BackgroundEarly introduction of food allergens into children's diet is considered as a strategy for the prevention of food allergy. The major fish allergen parvalbumin exhibits high stability against gastrointestinal digestion. We investigated whether resistance of carp parvalbumin to digestion affects oral tolerance induction.MethodsNatural Cyp c 1, nCyp c 1, and a gastrointestinal digestion‐sensitive recombinant Cyp c 1 mutant, mCyp c 1, were analyzed for their ability to induce oral tolerance in a murine model. Both antigens were compared by gel filtration, circular dichroism measurement, in vitro digestion, and splenocyte proliferation assays using synthetic Cyp c 1‐derived peptides. BALB/c mice were fed once with high doses of nCyp c 1 or mCyp c 1, before sensitization to nCyp c 1. Immunological tolerance was studied by measuring Cyp c 1‐specific antibodies and cellular responses by ELISA, basophil activation, splenocyte proliferations, and intragastric allergen challenge.ResultsWild‐type and mCyp c 1 showed the same physicochemical properties and shared the same major T‐cell epitope. However, mCyp c 1 was more sensitive to enzymatic digestion in vitro than nCyp c 1. A single high‐dose oral administration of nCyp c 1 but not of mCyp c 1 induced long‐term oral tolerance, characterized by lack of parvalbumin‐specific antibody and cellular responses. Moreover, mCyp c 1‐fed mice, but not nCyp c 1‐fed mice developed allergic symptoms upon challenge with nCyp c 1.ConclusionSensitivity to digestion in the gastrointestinal tract influences the capacity of an allergen to induce prophylactic oral tolerance.

Toxicological Examinations Following Oral Administration of Bacillus thuringiensis var kurstaki Biological Insecticide in Wistar Rats

Introduction: Bacillus thuringiensis var kurstaki is one of the best known biological insecticide which is used extensively in the world. This biopesticide has been broadly used against important insect pests and vectors of animals and humans. However, the controversial data was published to date considering toxicological studies on non-target species are urgent to determine probable adverse effects and risk assessment of this biopesticide. In this research, histopathological changes, hematological and some biochemical factors were evaluated following the single dose oral administration of B. thuringiensis in rats. Materials and Methods: Twelve Wistar rats were randomly divided into 2 groups including experimental and control groups. Animals were treated orally by gavage to single dose of sub-lethal dose (5000 mg/kg) of B. thuringiensis suspension. Finally, hematological factors (WBC, RBC, Hb, HCT, MCV, MCH, MCHC and PLT), some biochemical parameters (ALT, AST, ALP, BUN and Cr) and histopathological observations were evaluated. Results: The results demonstrated that oral administration of high dose of B. thuringiensis biopesticide induced some pathological complications including congestion and inflammation in vital organs of rats such as liver, heart, lung and kidney. It also caused hematological abnormalities and biochemical alterations in the experimental animals group. Conclusions: According to the findings, it can be concluded that high dose of B. thuringiensis biopesticide can induce toxicity in rats. Therefore, further investigations including subacute and chronic are recommended.