The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats / O óleo de alho, Allium sativum L. (Amaryllidaceae), como potencial protetor contra Anisakis spp. Infecção tipo II (L3) (Nematoda) em ratos Wistar

Abstract The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Resumo O consumo de peixe inadequadamente tratado termicamente representa um risco para a saúde pública, com a possibilidade da propagação de larvas de Anisakis. O presente estudo demonstrou as alterações fisiológicas e histopatológicas acompanhadas de inoculação oral de extractos brutos de Anisakis tipo II (L3) frescos e termicamente tratados em ratos. Os vermes foram isolados de um peixe marinho, examinados e identificados por microscopia de luz e eletrônica de varredura. O estudo foi conduzido em 6 grupos de ratos: controle (I), óleo de alho (GO) inoculado (II), L3 fresco inoculado (III), L3 tratado termicamente inoculado (IV), L3 fresco + GO inoculado (V), e um grupo L3 + GO tratado termicamente inoculado (VI). Observou-se que ratos inoculados com L3 fresco e tratados termicamente mostraram funções hepáticas e renais anormais, associadas à destruição da sua arquitetura normal. GO produziu um efeito protector em grupos de ratos inoculados com extractos L3 + GO através da melhoria das funções do fígado e dos rins, o que foi confirmado pela estrutura normal marcada da sua histologia. A cozedura de peixes infectados com L3 induziu alterações mais graves do que os peixes não cozidos. Recomenda-se a administração de alho antes e depois do consumo de peixe, para evitar o efeito perigoso dos anisakids, mesmo que sejam cozidos.

Preliminary study of Realgar and arsenic trioxide on gut microbiota of mice / 中国中药杂志

The aim of this paper was to observe the effect of Realgar and arsenic trioxide on gut microbiota. The mice were divided into low-dose Realgar group(RL), medium-dose Realgar group(RM), high-dose Realgar group(RH), and arsenic trioxide group(ATO), in which ATO and RL groups had the same trivalent arsenic content. Realgar and arsenic trioxide toxicity models were established after intragastric administration for 1 week, and mice feces were collected 1 h after intragastric administration on day 8. The effects of Realgar on gut microbiota of mice were observed through bacterial 16 S rRNA gene sequences. The results showed that Lactobacillus was decreased in all groups, while Ruminococcus and Adlercreutzia were increased. The RL group and ATO group were consistent in the genera of Prevotella, Ruminococcus, and Adlercreutzia but different in the genera of Lactobacillus and Bacteroides. Therefore, the effects of Realgar and arsenic trioxide with the same amount of trivalent arsenic on gut microbiota were similar, but differences were still present. Protective bacteria such as Lactobacillus were reduced after Realgar administration, causing inflammation. At low doses, the number of anti-inflammatory bacteria, such as Ruminococcus, Adlercreutzia and Parabacteroides increased, which can offset the slight inflammation caused by the imbalance of bacterial flora. At high doses, the flora was disturbed and the number of Proteobacteria was increased, with aggravated intestinal inflammation, causing edema and other inflammatory reactions. Based on this, authors believe that the gastrointestinal reactions after clinical use of Realgar may be related to flora disorder. Realgar should be used at a small dose in combination with other drugs to reduce intestinal inflammation.

Hydrogen sulfide protects H9c2 cardiomyoblasts against H2O2-induced apoptosis

Reactive oxygen species (ROS) are highly reactive chemical species that may cause irreversible tissue damage, and play a critical role in cardiovascular diseases. Hydrogen sulfide (H2S) is a gasotransmitter that acts as a ROS scavenger with cardio-protective effects. In this study, we investigated the cytoprotective effect of H2S against H2O2-induced apoptosis in cardiomyocytes. H9c2 rat cardiomyoblasts were treated with H2S (100 μM) 24 h before challenging with H2O2 (100 μM). Apoptosis was then assessed by annexin V and PI, and mitochondrial membrane potential was measured using a fluorescent probe, JC-1. Our results revealed that H2S improved cell viability, reduced the apoptotic rate, and preserved mitochondrial membrane potential. An increased Bcl-2 to Bax ratio was also seen in myocytes treated with H2S after H2O2-induced stress. Our findings indicated a therapeutic potential for H2S in preventing myocyte death following ischemia/reperfusion.

Properties of realgar bioleaching using an extremely acidophilic bacterium and its antitumor mechanism as an anticancer agent

Realgar is a naturally occurring arsenic sulfide (or Xionghuang, in Chinese). It contains over 90% tetra-arsenic tetrasulfide (As4S4). Currently, realgar has been confirmed the antitumor activities, both in vitro and in vivo, of realgar extracted using Acidithiobacillus ferrooxidans (A. ferrooxidans). Bioleaching, a new technology to greatly improve the use rate of arsenic extraction from realgar using bacteria, is a novel methodology that addressed a limitation of the traditional method for realgar preparation. The present systematic review reports on the research progress in realgar bioleaching and its antitumor mechanism as an anticancer agent. A total of 93 research articles that report on the biological activity of extracts from realgar using bacteria and its preparation were presented in this review. The realgar bioleaching solution (RBS) works by inducing apoptosis when it is used to treat tumor cells in vitro and in vivo. When it is used to treat animal model organisms in vivo, such as mice and Caenorhabditis elegans, tumor tissues grew more slowly, with mass necrosis. Meanwhile, the agent also showed obvious inhibition of tumor cell growth. Bioleaching technology greatly improves the utilization of realgar and is a novel methodology to improve the traditional method.

The Role of Hydrogen Sulfide in Acute Liver Injury Induced by Traumatic Stress in Rats / 法医学杂志

OBJECTIVE@#To explore the role of hydrogen sulfide (H2S) in acute liver injury induced by crushing hind limbs of rats.@*METHODS@#The rats were randomly divided into the following groups: control, crushing, H2S donor sodium hydrosulfide (NaHS) + crushing, H2S inhibitor propargylglycine (PAG) + crushing group. The acute liver injury model was established by 'crushing the hind limbs of rats with standard weight. Rats were sacrificed at 30 min and 120 min after the crush. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured by colorimetric method, and the content of H2S in plasma and the contents of malondialdehyde (MDA), protein carbonyl, glutathione (GSH) in the liver and the activity of H2S generating enzyme (cystathionine y-lyase, CSE) were determined by chemical method. The expression of CSE mRNA in liver was detected by RT-PCR.@*RESULTS@#For crush injury group, the levels of AST and ALT in serum, MDA and protein carbonyl in liver increased. The levels of GSH, CSE, CSE mRNA in liver and H2S in serum decreased. The administration of NaHS before limbs crush could attenuate the changes of liver injury, but the pre-treatment with PAG could exacerbate the changes.@*CONCLUSION@#The decrease of H2S production could involve in mediating the acute liver injury induced by traumatic stress in rats.

Caspase-8 and p38MAPK in DATS-induced apoptosis of human CNE2 cells

Nasopharyngeal carcinoma is a common malignancy in Southern China of uncertain etiologic origin. Diallyl trisulfide (DATS), one of the major components of garlic (Allium sativum), is highly bactericidal and fungicidal. In this study, we investigated the function of p38 mitogen-activated protein kinase (MAPK) and caspase-8 in DATS-induced apoptosis of human CNE2 cells using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], flow cytometry assay, and Western blotting. After CNE2 cells were treated with DATS (50, 100, or 150 μM) for 24 h, cell viability rates were 75.9, 63.4 and 39.6 percent, and apoptosis rates were 24.5, 36.9, and 62.4 percent, respectively. The data showed that DATS induced CNE2 cell death in a dose-dependent manner. After human CNE2 cells were treated with 100 μM DATS and inhibitors (10 μM SB203580 and Z-LETD-FMK for p38MAPK and caspase-8, respectively), changes in cell viability and apoptosis and in p38MAPK and caspase-8 activity were detected. Cell viability rates were 66.5 and 68.1 percent and decreased 9.9 and 11.5 percent compared with inhibitor treatment alone. Apoptosis rates were 31.53 and 29.98 percent and increased 9.1 and 10 percent compared with inhibitor treatment alone. The results indicated that DATS activates p38MAPK and caspase-8, but both inhibitors have an effect on P38MAPK and caspase-8 activity. In conclusion, our data indicate that p38MAPK and caspase-8 are involved in the process of DATS-induced apoptosis in human CNE2 cells and interact with each other.

Alternative control of Tetranychus evansi Baker & Pritchard (Acari: Tetranychidae) on tomato plants grown in greenhouses

Tetranychus evansi Baker & Pritchard is an important pest of solanaceous plants, including tomatoes. This mite is characterized by a high reproductive rate, which leads to high population growth in a short period of time causing important economic damage. Control of T. evansi is mainly through synthetic acaricides. In searching for environmentally friendly control measures, we evaluated the efficiency of alternative products to control T. evansi on tomato plants under greenhouse conditions. The products tested were lime sulphur and neem based products. We first estimated the lethal concentration (LC) and instantaneous rate of increase (r i) of T. evansi exposed to different product concentrations in laboratory conditions, and later tested the efficacy of LC95 and the concentrations that restrained mite population growth (r i = 0) in greenhouse conditions. The following treatments were repeated three times: NeemPro (81.0 and 71.6 mg a.i./l), Natuneem (31.1 and 20.4 mg ai/l), Organic Neem (39.1 and 30.4 mg a.i./l), lime sulphur (1.0 and 0.6 percent) and water (control). For all products, control provided by LC95 was higher than provided for lower concentrations (r i = 0) one day after spraying. However, after five days, for both concentrations, the percentage of T. evansi population reduction was superior to 95 percent and increased over time. Only plants sprayed with Natuneem (31.1 mg a.i./l) showed symptoms of phytotoxicity. Lime sulphur and neem based products, applied in appropriate concentrations and formulations, bear out as a viable alternative to control T. evansi on tomato plants.

Implications of dibenzyl trisulphide for disease treatment based on its mode of action / Implicaciones del trisulfuro de dibencilo para el tratamiento de enfermedades a partir de su modo de acción

Studies conducted on the secondary metabolite (natural product), dibenzyl trisulphide (DTS), which was isolated from the sub-tropical shrub Petiveria alliacea (guinea hen weed, anamu) [Phytolaccaceae] have shown tremendous pharmaceutical promise as a drug prototype. This is now reflected in the development of the broad spectrum anti-cancer molecule, fluorapacin (bis(4-fluorobenzyl) trisulphide) which has an excellent safety profile. The mode of action elucidated for DTS is the mitogen activated protein extracellular regulated kinases 1 and 2 (MAPKinases ERK 1 and ERK 2). The MAPKinase signal transduction biochemical pathways are important in the regulation of a wide range of cellular processes which are important in disease establishment. These processes include: cancer cell proliferation, nerve repair, memory enhancement, autoimmune diseases, which are linked to thymic cell involution and bone marrow functions, cerebrovascular and cardiovascular diseases. In addition to the MAPkinase signal transduction mode of action, DTS also prevents the denaturation of serum albumin which is a feature of nonsteroidal anti-inflammatory drugs, thus supporting the molecule's possible role in the treatment of inflammatory ageing diseases.

Los estudios realizados sobre el metabolito secundario (producto natural), trisulfuro de dibencilo (TSD), que fue aislado del arbusto subtropical Petiveria alliacea (hierba de guinea, anamú) [Phytolaccaceae] muestran que se trata de una tremenda promesa farmacéutica como prototipo de droga. Esto se refleja actualmente en el desarrollo de la molécula anticancerígena de amplio espectro, la fluorapacina (bis (4-fluorobencilo) trisulfuro) que posee un excelente perfil de seguridad. El modo de acción para el TSD se explica partiendo de las proteínas quinasas 1 y 2 activadas por mitógenos y reguladas por señales extracelulares (Quinasas MAP ERK 1 y ERK 2). Las vías bioquímicas de transducción de la señal de la quinasa MAP, son importantes en la regulación de una amplia variedad de procesos celulares, importantes a la hora de determinar una enfermedad. Dichos procesos comprenden: la proliferación de la célula cancerosa, la reparación de nervios, el mejoramiento de la memoria, y las enfermedades autoinmunes, vinculadas con la involución tímica y las funciones de la médula, las enfermedades cerebrovasculares y cardiovasculares. Además del modo de acción de las transducción de señales de la quinasa MAP, el TSD previene también la desnaturalización de la albúmina sérica, lo cual constituye una característica de las drogas anti-inflamatorias no esferoidales, apoyando así el posible papel de las moléculas en el tratamiento de las enfermedades inflamatorias en el proceso de envejecimiento.

A critical review of the therapeutic potential of dibenzyl trisulphide isolated from Petiveria alliacea L (guinea hen weed, anamu) / Un estudio crítico del potencial terapéutico del trisulfuro de dibencilo aislado a partir de petiveria alliacea l (yerba de guinea, anamú)

The data compiled in the present review on dibenzyl trisulphide (DTS) isolated from Petiveria alliacea L (the guinea hen weed or anamu) revealed that the compound and its derivatives could be of tremendous pharmaceutical interest. The mode of action elucidated for DTS revealed that it is a mitogen activated protein extracellular regulated kinases 1 and 2 (MAPKinases erk1 and erk 2) signal transduction molecule. Dibenzyl trisulphide caused hyper-phosphorylation of growth factor induced MAPKinases (erk 1 and erk 2) phosphorylation, a process critical for the improvement of long term memory, and is implicated in neuronal growth. Dibenzyl trisulphide and its derivatives exhibited potent anti-proliferation/cytotoxic activity on a wide range of cancer cell lines. The cytotoxic activity of DTS was increased by 70-1000 fold when bound to albumin in vitro. Dibenzyl trisulphide seems to have a cytokine switching mechanism in which it down regulates cytokines from the Type I helper cells (Th -1 cell) pathway which contained several pro-inflammatory cytokines and up-regulates those on the Type 2 helper cells (Th-2) pathway. The trisulphide up-regulates some reticuloendothelial system parameters eg granulocyte counts and increased thymic and Peyer's patches masses via cell proliferation processes which are known to be regulated via the MAPKinase signal transduction pathway. When the zygotes ofAsternia pectinifera (Starfish) were exposed to DTS at concentration of 10 mM, a dose lethal to all cancer cells tested, it was observed that the sensitive process of protein biosynthesis was not affected Similarly, the proliferation of the HOFA human fibroblast, a noncancerous cell line, was not severely affected by DTS at 8.9 microM over seven days, a concentration also lethal to most cancer cell lines tested The implications of the findings will be highlighted in the present review.

Los datos compilados en el presente estudio sobre el trisulfuro de dibencilo (TSD) aislado a partir de Petiveria alliacea L (yerba de Guinea, ó anamú) revelaron que el compuesto y sus derivados podrían tener extraordinario interés farmacéutico. El modo de acción esclarecido en el TSD, reveló que se trata de una molécula de transducción de señales de proteínas kinasas 1 y 2 (MAP quinasas ERk 1 y 2) reguladas extracelularmente y activadas por mitógenos. El trisulfuro de dibencilo causó hiperfosforilación de la fosforilación de las quinasas MAP (Erk 1 y 2) inducidas mediante factor de crecimiento, un proceso crítico para el mejoramiento de la memoria a largo plazo, y que está implicado en el crecimiento neuronal. El trisulfuro de dibencilo y sus derivados mostraron una poderosa actividad citotóxica y antiproliferativa en una amplia gama de líneas celulares de cáncer. La actividad citotóxica del TSD se incrementaba de 70 á 1000 veces, cuando se vinculaba a la albúmin in vitro. El trisulfuro de dibencilo parece poseer un mecanismo conmutador citoquínico que regula por decremento las citoquinas provenientes de la vía de las células auxiliares de tipo 1 (células Th-1), que contiene varias citoquinas pro-inflamatorias y regula por incremento las de la vía de las células auxiliares de tipo 2 (Th-2). El trisulfuro regula por incremento los parámetros del sistema reticuloendotelial, p.ej. los conteos de granulocitos y el aumento tanto de las masas tímicas como de las masas de placas de Peyer, a través de los procesos de proliferación celular, de los cuales se sabe que son regulados mediante la vía de la transducción de señales de la quinasa MAP. Cuando los cigotos de Asternia pectinifera (estrella de mar) fueron expuestos al TSD a una concentración de 10 mM ­ una dosis letal para todas las células cancerosas sometidas a prueba ­ se observó que el proceso sensible de biosíntesis de la proteína no era afectado. De modo similar, la proliferación del fibroblasto humano HOFA ­ una línea celular no cancerosa ­ no fue afectada severamente por el TSD a 8.9 µM en siete días ­ una concentración letal para la mayoría de las líneas celulares cancerosas sometidas a prueba. Las implicaciones de los hallazgos se pondrán de relieve en el presente estudio

Involvement of multiple signaling pathways in diallyl sulfide mediated apoptosis in mouse skin tumors.

Many chemopreventive agents appear to target signaling intermediates in apoptosis-inducing pathways. Inherently, the process of neoplastic conversion selects against apoptosis to initiate, promote, and perpetuate the malignant phenotype. Thus, targeting apoptosis pathways in pre-malignant cells, in which these pathways are still relatively intact, may be an effective module of cancer prevention. Diallyl sulfide (DAS), a naturally occurring organosulfide, present in garlic, is reported to have pleiotropic biological effects. DAS is known to inhibit chemically induced tumors in a number of in vivo and in vitro studies. The aberration of tumor suppressor gene, p53 and the ras oncogene have been linked to the induction of multiple signaling pathways and to the resistance offered by cancer cells to the apoptosis. Therefore, the present study was carried out to investigate the role of DAS on modulation of multiple p53 and ras-induced signaling pathways in 7,12-dimethylbenathacene (DMBA) induced skin carcinogenesis. The results showed that DAS up regulates expression of tumor suppressor protein p53 (wt p53) and its downstream target molecule p21/waf1. Proapoptotic protein, bax was upregulated by DAS supplementation. An opposite trend was observed in DMBA induced antiapoptotic proteins expressions, survivin and bcl-2, which were significantly downregulated by DAS supplementation. In the present study we also demonstrated that DAS supplementation significantly reduces the expression of ras oncoprotein and to modulate expression of its signaling molecules including PI3K/Akt and MAPKs. Western blot analysis demonstrated that DAS significantly reduced the DMBA induced protein expressions of PI3K/Akt and p38MAPK. However, DAS supplementation did not alter the expression JNK1 and ERK1/2. Thus, our results confirm that DAS can adopt a multi-prong strategy to target multiple signaling pathways leading to induction of apoptosis and inhibition of growth of DMBA induced skin tumors in Swiss albino mice. Although studies of single pathways have been helpful in guiding investigations, new tools to study the integration and multiplicity of signaling pathways hold the hope of improved understanding of the signaling pathway alterations in cancer chemoprevention by naturally occurring compounds.

In vitro anti-proliferation/cytotoxic activity of sixty natural products on the human SH-SY5Y neuroblastoma cells with specific reference to dibenzyl trisulphide

Sixty natural products belonging to the following structural classes: artemisinins, coumarins, flavonoids, tannins, tetrahydroberberine alkaloids, tetracyclic triterpenes, tetranortriterpenoids and polysulphides were screened against the human SH-SY5Y neuroblastoma cell line revealing differences in their effects on cell morphology and in anti-proliferation/cytotoxic activity. Based on the data obtained, dibenzyl trisulphide is the most effective anti-proliferative/cytotoxic compound. In addition, we hereby propose the human SH-SY5Y cell line as a sensitive and uncomplicated in vitro test system for detecting compounds with potential anti-proliferation/cytotoxic activity

Bioassay of three sulphur containing compounds as rat attractant admixed in cereal-based bait against Rattus rattus Linn.

Three sulphur containing compounds, carbon disulphide, dimethyl disulphide and dimethyl sulphide were bioassayed for preference after admixing them in cereal base as ready bait block for use against commensal rat, R. rattus (wild type) in four way choice chamber system. Rat preference for different baits was also studied with automatic recording animal activity meter. Rats exhibited attractancy to the baits at 0.005% concentration of all the three compounds while at 0.01% concentration they have showed repellency. Dimethyl sulphide at 0.005% concentration showed better attractancy towards both sexes of rat.

Effects of allyl sulfur compounds and garlic extract on the expression of Bcl-2, Bax, and p53 in non small cell lung cancer cell lines

Allyl sulfur compounds play a major role in the chemoprevention against carcinogenesis. The present study compared the antiproliferative effects of diallyl sulfide (DAS), diallyl disulfide (DADS) and garlic extract on p53-wild type H460 and p53-null type H1299 non small cell lung cancer cells (NSCLC). The DAS and DADS treatment of both H460 and H1299 cells resulted in the highest numbers of cells in apoptotic state as measured by acridine orange staining, however, garlic extract treatment did not induce any significant apoptotic cells by MTT assay. DADS was found to be more effective in inducing apoptosis on NSCLC. The level of p53 protein in H460 cell was increased following DADS treatment. DAS and garlic extract treatment of H460 cells induced a rise in the level of Bax and a fall of Bcl-2 level. These results demonstrate that DAS, DADS and garlic extract are effective in reduction of anti-proliferative gene in NSCLC and suggest that modulation of apoptosis-associated cellular proteins by DAS, DADS and garlic extract may be the mechanism for apoptosis which merit further investigation as potential chemoprevention agents.

Modification of radiation induced changes in murine hepatic lipid profiles by garlic (Allium sativum Linn.) unsaturated oils.

Adult male Swiss albino mice were administered 74 kBq g-1 body weight of 45Ca in the presence and absence of garlic unsaturated oils, and the changes in total lipids, triglycerides, phospholipids and free fatty acids contents of liver were observed at various intervals from 1 to 14 days post-treatment. The results obtained indicate that garlic oils prevented rapid increase in hepatic total lipids, triglycerides and phospholipids and decrease in free fatty acids induced by radiocalcium and the values reached normal values earlier in garlic treated animals than in irradiated animals. Possible mechanism underlying the protective action of garlic oils is reported.

Hypolipidemic effect of garlic protein substituted for casein in diet of rats compared to those of garlic oil.

Garlic (Allium sativum Linn.) is ascribed with many therapeutic effects. For the present study, the water soluble proteins and the essential oil of garlic were investigated for their hypolipidemic effect on hyperlipidemia induced by cholesterol containing diet in albino rats. Both garlic protein (16% of diet) and garlic oil (100 mg/kg body weight/day) exhibited significant lipid lowering effects. The hypolipidemic action is primarily due to a decrease in hepatic cholesterogenesis in the treated rats. Even though garlic oil was found to be more effective, the garlic protein is more palatable and free from an obnoxious smell.

Biochemical effects of garlic protein diet and garlic oil on glycosaminoglycan metabolism in cholesterol fed rats.

Garlic protein diet or daily administration of garlic oil to 2% cholesterol fed rats controlled significantly the increases in sulphated glycosaminoglycans in their heart and aorta. However hyaluronic acid level increased. UDPG dehydrogenase decreased and several degrading enzymes increased in the aorta on treatment. The effects of treatment were just the reverse in liver. The high percentage of cysteine in garlic protein and the reactive disulphide group in the oil may be responsible for their beneficial effects.