Inhibition of foot and mouth disease virus (FMDV) uncoating by a plant-derived peptide isolated from Melia azedarach L leaves

Summary  Meliacine (MA), a peptide isolated from leaves of the high plant Melia azedarach L inhibited the multiplication of foot and mouth disease virus (FMDV) in BHK-21 cells. In this report, we establish that the MA-inhibitable process takes place within the first hour of the viral reproductive cycle. MA had no virucidal effect and did not affect adsorption and penetration of the virus in cells. In experiments with neutral red-labeled virus, it was found that MA significantly suppressed the development of photoresistance of the virus in infected cells. In untreated cultures nearly all virus which adsorbed to cells was uncoated within 1 h at 37 °C, whereas in treated cultures, even after 3 h only 3% of the virus was uncoated. Labeling of BHK-21 cells with acridine orange showed that MA affects the pH of intracellular acidic vesicles. Therefore, it is concluded that MA prevents the process of uncoating of FMDV in BHK-21 cells by inhibiting vacuolar acidification. Accepted October 15, 1997 Received April 21, 1997     

Ovicidal and larvicidal activity of crude extracts of Melia azedarach against Haemonchus contortus (Strongylida)

The rapid development of anthelmintic resistance, associated with the high cost of the available anthelmintic drugs, has limited the success of gastrointestinal nematodiosis control in sheep and goats and thus created interest in studying medicinal plants as an alternative source of anthelmintics. The aim of this study was carried out to evaluate the anthelmintic activity of the leaves and seed aqueous and hydro-alcoholic extracts of Melia azedarach L. (Meliaceae) were tested for in vitro ovicidal and larvicidal activity against Haemonchus contortus (Strongylida). Both extracts were evaluated at five concentrations: 12.5, 6.2, 3.12, 1.56, and 0.78 mg/ml. The leaves aqueous and hydro-alcoholic extracts inhibited 99.4% and 100% of the egg hatching and 100% of larval development at 12.5 mg/ml, respectively. In a similar way, the leaves hydro-alcoholic extract was the most active on egg inhibition (ED ₅₀ = 1.97 and ED ₉₀  = 5.05 mg/ml), leaves and seed aqueous and hydro-alcoholic extracts showed the best inhibition of larval development (ED ₅₀  = 3.01, 2.43, 3.17, 2.40, and ED ₉₀  = 10.53, 8.14, 11.94, and 8.19 mg/ml), respectively. These results suggest that utilization of M. azedarach extracts is useful in the control of H. contortus.     

Effects of Azadirachta indica and Melia azedarach (Meliaceae) extracts from leaves on Trypanosoma cruzi growth and ultrastructure

The chloroformic extracts from dried fresh leaves ofAzadirachta indica A. Juss. and Melia azedarach L. (Meliaceae) showed marked inhibitory activity on epimastigotes growth of Trypanosoma cruzi, evidenced by 96-wells microtiter plate bioassay and radioactive thymidine incorporation experiment. Each chloroformic extract was separated using silica gel and alumina column. In transmission electron microscopy the bioactive chromatographic fractions caused ultrastructural changes in epimastigotes such as vacuolization probably induced by degeneration of the kinetoplast-mitochondrion complex, organelle degeneration, and cell division disruption. In spectral analysis these bioactive fractions seemed to be composed mainly of fatty acid mixtures.     

In vitro antimalarial activity of extracts of Albizia gummifera,Aspilia mossambicensis,Melia azedarach and Azadirachta indica against Plasmodium falciparum

Since chemotherapy is presently the primary strategy of malaria control in the world, and some malaria parasites are developing resistance to the commonly used antimalarial drugs, new antimalarial compounds are required. Therefore, it is important to test antimalarial activities of medicinal plant extracts which most herbalists claim to cure malaria. We evaluated the antimalarial activities of extracts of Albizia gummiffera, Aspilia mossambicensis, Melia azedar and Azadirahchta indica against laboratory adapted isolates of Plasmodium falciparum using an in vitro radioisotopic uptake technique. Chloroquine was used as a reference antimalarial drug. Al. gummifera had the highest antimalarial activity (mean fifty percent inhibitory concentration {IC(50)S} in ug/ml of test culture =3.5 +1.6SD, n=3) followed by As. mossambicensis (mean IC(50)=29.3+11.8SD, n=4) and Me. Azedarach (mean IC(50) =299.7+202.0SD, n=4). And lastly Az. Indica (mean IC(50)=349.9+213.1 SD, n=4). The antimalarial activities of the reference drug, chloroquine, was far much higher (mean IC(50)=0.065+0.057SD, n=)4). These findings show that Al. gummifera and As. mossambicensis plant extracts have potent antimalarial compounds. Phytochemical analyses should be done on these two plants to isolate the compound(s) containing he active principles(s).     

Outpatient percutaneous treatment of deep venous malformations using pure ethanol at low doses under local anesthesia

INTRODUCTION:

Venous malformations are the most frequent vascular malformation. Deep venous malformations are located in subcutaneous tissue or in the muscles. Percutaneous sclerotherapy is the treatment of choice, and the use of ethanol at low doses has not yet been described.

OBJECTIVE:

To analyze the results of treating Deep venous malformations patients with low doses of ethanol.

METHODS:

Thirty‐nine patients treated between July 1995 and June 2007 were followed up prospectively over a median period of 18 months. Twenty‐nine were female (74.4%) and 10 were male (25.6%), with ages ranging from 11 to 59 years (median of 24 years). All of the lesions affected limbs, and the main symptom reported was pain (97.4%). Each patient underwent fortnightly alcohol application sessions under local anesthesia on an outpatient basis. The lesions were classified into three groups according to size using nuclear magnetic resonance imaging: small, up to 3 cm (4 patients); medium, between 3 and 15 cm (27 patients); and large, greater than 15 cm (8 patients).

RESULTS:

The symptoms completely disappeared in 14 patients (35.9%) and improved in 24 (61.5%). The lesion size reduced to zero in 6 patients (15.4%) and decreased in 32 (82%). The median number of sessions was 7. There were no complications in 32 patients (82%), while 3 presented local paresthesia (7.7%), 2 superficial trombophlebites (5.1%), 1 skin ulcer (2.6%), and 1 case of hyperpigmentation (2.6%).

CONCLUSION:

Outpatient treatment for Deep venous malformations patients using ethanol at low doses was effective, with a low complication rate.     

Hemodialysis as a treatment of severe ethanol poisoning

A 64-year-old woman presented with coma and shock due to severe ethanol intoxication. Her initial, markedly elevated blood alcohol level of 136.5 mM fell only by 16% after a 4-hour period of conservative treatment consisting of mechanical respiration and the administration of intravenous fluids, vasopressors and inotropics. Subsequent hemodialysis rapidly reduced her blood ethanol concentrations to less threatening levels, with prompt restoration of her consciousness. Hemodialysis may be life-saving and should be considered in patients with severe ethanol intoxication.     

Posttrial treatment with ethanol enhances recall of prose narratives

A small number of studies have examined the effects of postlearning alcohol intoxication on memory for various materials. In contrast to most research examining the effects of alcohol on human memory, these studies demonstrated a facilitation of memory for information learned prior to intoxication. The present study was designed to examine the effects of alcohol on memory for two different kinds of materials. Standard word lists and narrative prose passages were employed to obtain a more detailed picture of the effects of posttrial intoxication with alcohol on memory. Intoxication with alcohol did not affect subject's ability to recall simple word lists. However, recall of prose passages was facilitated to a statistically significant degree. Results are discussed within the context of a current theory of retrograde facilitation of memory via various drugs/substances.     

Inhibitory effect of Crocus sativus L. ethanol extract on adjuvant-induced arthritis

Saffron is a well-known spice produced from dried stigmas of Crocus sativus L. flowers. Apart from its wide use in food preparations, it also has a broad range of medical properties. We examined the potential anti-inflammatory effects of saffron ethanolic extract (SEE) using an animal model of arthritis. Adjuvant-induced arthritis was induced in Wistar rats by injection of Complete Freund's Adjuvant. The rats were then injected intraperitoneally every other day with 25–600 mg SEE/kg or dexamethasone (DEX, 2 mg/kg). Changes in body weight, paw oedema and arthritis indices were recorded over the subsequent 12 days of treatment. Results revealed that SEE particularly at the higher concentrations significantly reduced paw and tibiotarsal joint diameters and comparing with DEX caused no significant change in body weight. These observations suggest that SEE displays a considerable anti-inflammatory potency and could potentially be used as an anti-arthritic agent in control of inflammation in rheumatoid arthritis.     

Increased systemic and brain cytokine production and neuroinflammation by endotoxin following ethanol treatment

Background  

Cytokines and alcohol share a common modulation of inflammation and hormones as well as being implicated in multiple diseases, but the mechanisms are poorly understood. The purpose of this study was to investigate the interaction of liver, serum and brain cytokines as well as whether ethanol would potentiate endotoxin (Lipopolysaccharide, LPS) responses once ethanol had cleared.     

The relationship between apoptosis and splenocyte depletion in rats following ethanol treatment

Splenocyte depletion observed in chronic ethanol-treated rats (ETRs) was studied in relation to apoptosis. The rats were fed with ethanol in a Liber-DeCarli liquid diet (36% of total calories as ethanol) for 7 weeks. Spleens of ETRs and control rats were examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method, immunohistochemistry using anti-rat p53 and RM4 (specific for macrophages) monoclonal antibodies, and transmission electron microscopy (TEM). The splenic white pulp in ETRs decreased in size and showed a moth-eaten appearance because of the severe depletion of splenocytes. Most TUNEL-positive cells aggregated into clusters or nests and were not isolated in the white pulp of ETRs. The site of RM4 immunoreactivity was consistent with that of clusters of TUNEL-positive cells. The p53 immunoreactivity was observed in apoptotic splenocytes that were isolated or phagocytosed by macrophages. TEM study revealed the increase in tingible body macrophages phagocytosing apoptotic splenocytes in their cytoplasm in ETRs. Chronic ethanol intake certainly induces apoptosis in splenic white pulps, and tingible body macrophages act as both sentinels and scavengers of apoptotic splenocytes expressing p53.     

乙醇处理与大鼠脑内多巴胺能体系的关系

目的观察乙醇处理大鼠脑内酪氨酸羟化酶(TH)和多巴胺转运体(DAT)的表达变化,判断乙醇对脑内多巴胺(DA)能体系的影响。方法选取Wistar大鼠60只,随机分为对照组和乙醇处理组,每组30只,乙醇处理组大鼠以20%乙醇代替饮水饲养6个月;利用免疫组织化学、流式细胞术及免疫印迹等方法,分析乙醇处理大鼠有关脑区TH和DAT的表达改变。结果1.免疫细胞化学法研究发现,乙醇处理组大鼠脑内黑质(SN)-腹侧被盖区(VTA)、尾壳核(CPu)和伏核(NACC)TH的灰度值明显低于对照组(P0.05);尾壳核和伏核DAT的灰度值明显低于对照组(P0.05)。2.流式细胞术检测发现,乙醇处理组大鼠中脑TH的表达量明显高于对照组(P0.05)。3.免疫印迹检测发现,乙醇处理组大鼠所观察脑区TH和DAT与β-actin条带相对吸光度比值明显高于对照组(P0.05)。结论乙醇处理增加大鼠脑内TH和DAT的表达。     

CT导向下经皮无水乙醇消融在复杂性肾癌治疗中的临床应用

目的 探讨CT导向下经皮无水乙醇消融(PEA)在复杂性癌治疗中的临床价值.方法 对7例复杂性肾癌患者,其中孤立肾肾癌2例,单侧肾肾癌5例,病灶数11个,肿瘤直径1.7～8.4 cm,在CT引导、局麻下进行经皮穿刺无水乙醇消融治疗,常规行螺旋CT平扫和增强扫描进行随访评价疗效.结果 患者平均每人2次PEA治疗,随访12～26个月,中位时间15个月.1例孤立肾肾癌患者经2次PEA治疗后,已随访1年9个月未见复发;1例孤立肾多发肾癌患者4次PEA治疗后肾衰死亡;2例肾癌伴腰痛血尿症状经PEA治疗后病灶消融完全,血尿症状缓解;其他3例肾癌患者分别随访1年6个月、2年2个月和1年,1例死于远处转移,其他2例CT复查未见肿瘤进展.全部患者无1例出现尿瘘、出血、肠穿孔或针道转移等并发症.结论 CT导向下无水乙醇消融治疗对复杂性肾癌是一种微创、疗效确切、可供选择的局部治疗方法 ,但对于孤立肾肾癌,PEA治疗须谨慎.     

Effect of ethanol and cocaine treatment on the immune system of v-Ha-ras-transgenic mice

The objective was to investigate if the presence of the v-Ha-ras oncogene could induce immune changes different to the ones observed in normal mice. Therefore, we decided to use Oncomice, the transgenic mice with an activated v-Ha-ras oncogene under the control of the mouse mammary tumor virus-promoter, and their normal inbred counterparts, FVB mice. Both strains of mice were fed the Lieber-DeCarli liquid diet with ethanol or the isocaloric control diet and were injected daily with cocaine or saline. The percentage and absolute number of T and B lymphocytes in the spleen and thymus were determined. The in vitro production of TNF-α (tumor necrosis factor-α), IL-2 (interleukin-2) and IFN-γ (interferon-gamma) by spleen cells, and the levels of serum sIL-2R (soluble IL-2 receptor) were also measured. Oncomice fed the Lieber-DeCarli ethanol diet or receiving either saline or cocaine injections presented a higher tumor incidence than Oncomice receiving the control diet. A reduced total number of thymocytes as well as absolute number of cells in all the subsets was found in Oncomice. Moreover, a decreased percentage of CD8⁺ cells was also observed in Oncomouse spleens. These features were even more marked in ethanol-treated Oncomice. Higher serum sIL-2R levels were observed in Oncomice, especially in mice treated with ethanol or cocaine. The results suggest that the oncogene product, P21^ras, plays an important role in dysregulating the immune system and hence in favoring tumorigenesis.     

Study of ethanol induced toxicity in liver explants using microfluidic devices

Current in vitro methodologies for the culture and analysis of liver specific responses lack the sophistication of in vivo dynamics. In this work, a microfluidic based experimental methodology has been utilized to reproduce a biomimetic microenvironment in which pseudo in vivo liver tissue studies can be carried out under in vitro conditions. This innovative technique, which exploits the inherent advantages of microfluidic technology, has been utilised to study the viability and functionality of explant liver tissue over four days in the presence of varying concentrations of ethanol. Concentrations of ethanol as low as 20 mM have produced a decrease in WST-1 metabolism, a marker of mitochondrial activity, and an increase lactose dehydrogenase release, reflecting cell death, in the explant samples; these effects increase with higher ethanol concentrations. A concomitant decrease in albumin and urea synthesis was also observed. We believe the proposed methodology is widely applicable and is clearly of relevance to biological and clinical research including drug development and toxicity, as well as enabling better fundamental understanding of tissue/cell processes.     

天麻素对乙醇诱导肝细胞株L02细胞损伤的影响

目的:探讨天麻素对乙醇诱导肝细胞株L02损伤的影响。方法:将人肝细胞株L02培养,加入不同浓度的乙醇,采用MTT比色法确定乙醇作用的浓度。用流式细胞仪测定细胞活性氧簇(ROS)及线粒体膜电位的改变;采用反相高效液相色谱(RP-HPLC)分析法检测细胞ATP含量。结果:乙醇(25 mL/L)作用36 h可损伤肝细胞,细胞内ROS的水平明显增加;而细胞线粒体膜电位及肝细胞中ATP的含量则显著降低(P0.01)。天麻素具有减轻细胞损伤的作用,并可降低损伤的肝细胞内ROS的含量,增加细胞线粒体膜电位和ATP的水平。结论:天麻素可抑制乙醇诱导的肝细胞损伤及脂质过氧化反应,改善线粒体功能,增加能量合成,发挥保护肝细胞的作用。     

Combining diagnosis and treatment using ASBRU

Traditionally, diagnosis and treatment have been seen as two distinct tasks. Consequently, most approaches to computer supported health care focus on one of the two-mostly on diagnosis or rather on the interpretation of measurements which is much better understood and formalised. However, in practice diagnosis and treatment overlap and influence each other in many ways. Combinations range from repeatedly going through the diagnosis-treatment loop over a period of time to permanent monitoring of the patients' health condition as it is done in intensive care units. In this article we describe how to model these combinations using the clinical protocol-representation language ASBRU. It implements treatment steps in a hierarchy of skeletal, time-oriented plans. Diagnosis can either be described in a declarative way in the conditions, under which treatment steps are taken or it can be modelled explicitly as plans of their own right. We demonstrate our approach using examples taken from the American Association of Paediatricians' guideline for the treatment of hyperbilirubinemia in the new-born.     

Neural tube defects in early rat embryos following maternal treatment with ethanol and caffeine

Alcohol and caffeine consumption during pregnancy are risk factors for fetal outcome. Their combined effects on the early development of the central nervous system were investigated in the rat. Neuralation was severely affected, resulting in a wide spectrum of developmental defects.     

The effects of ethanol and silymarin treatment during gestation on spatial working memory.

Background  

Using a rat model we have found that the bioflavonoid silymarin (SY) ameliorates some of the negative consequences of in utero exposure to ethanol (EtOH). In the current study our aim was to determine if spatial working memory (SWM) was impaired in offspring whose mothers were maintained on a liquid diet containing EtOH during different gestational weeks. We also determined if SWM was altered with a concomitant administration of SY with EtOH during specific gestational weeks.