The proliferating nuclear fraction as a prognostic factor in superficial bladder carcinomas

Pongamia pinnata root has been advocated in Ayurveda for treatment of various inflammatory and infective conditions including ulcers. Sequential petroleum ether, benzene, chloroform, acetone and ethanolic extracts of P. pinnata roots when administered in the dose of 50 mg/kg, i.p. in rats was found to have anti-inflammatory, analgesic activity while pentobarbitone-induced 'sleep time' was reduced by all the extracts except petroleum ether which, however, enhanced it. They were also found to possess antiulcer effects when administered either by i.p. (45 min before) or oral route (45 min before or for 4 days) against restraint-stress or pylorus-ligated gastric ulcers in rats, the maximum protection being afforded by petroleum ether and ethanol extracts. The mechanism of antiulcer effect could either be due to decrease in acid-pepsin secretion and augmentation of mucin secretion as observed with ethanol extract, while petroleum ether extract might be producing the effect by virtue of its anti-stress activity.     

Staging of Hodgkin''s disease: a surgical perspective

1. The preliminary phytochemical screening of the two seeds established the presence of carbohydrates and/or glycosides, flavnoids, unsaturated sterols and/or triterpenes, saponins, trypsin inhibitors and haemagglutinins. In addition, it established the absence of cardenolides, tannins, alkaloids and oxidase enzyme. 2. Certain pharmacopoeial constants, including moisture, ash, acid-insoluble ash, water-soluble ash and crude fibre were determined. 3. The two seeds were subjected to successive extractions with different organic solvents such as petroleum ether (50-70 degrees C), diethyl ether, chloroform and ethyl alcohol. The successive yields of extractives were determined. Examination of the crude extracts showed that petroleum ether extract contained sterols and/or triterpenes, while ether, chloroform, and ethyl alcohol extracts contained reducing substances. 4. General analysis of the two seeds for proteins, fats, carbohydrates, fibre and ash contents were carried out and the results were given in g/100 g dry seeds. Pigeon pea contained 25.2 g protein, 170 mg calcium and 8.9 mg iron. The protein content of kidney bean was 23 g, while calcium and iron contents were 134 mg and 8.02 mg respectively. 5. Extractions of the proteins using different solvents such as cold water, hot water, saline buffer pH 7 and sodium hydroxide was the best extractant. 6. The amino-acid content of the two seeds, whether raw or cooked, showed that they were deficient in methionine, cystine and tryptophan. Other essential amino acids were present in amounts higher than that given by the FAO provisional pattern. 7. Cooking the seeds by the popular methods used in the country resulted in an increase in the amounts of the amino acids, threonine, leucine and isoleucine, while the other amino acids present remained unchanged or decreased. It was also observed that cooking the seeds destroyed the trypsin inhibitors and haemagglutinins found in the two seeds.     

The beginnings of anesthesia

In the nineteenth century, introduction of the first inhaled general anaesthetic (Long, 1842) induced a further search for new types of anaesthesia: carbon dioxide, petroleum ether, derivatives of ethylene, acetone, methyl dichloride, and the study of a new technique-hypnosis. Only chloroform, ether, nitrous oxide were used. Towards the end of the century, ether became the mainstay of inhaled anaesthetics. The other routes of administering anaesthetics (rectal, venous, spinal, local) appeared around 1860.     

Chemical constituents of Rosa laevigata Michx

Six compounds were isolated from Rosa laevigata. Five of them were obtained from the ethanolic extract and identified as 2 alpha, 3 beta, 19 alpha, 23-tetrahydroxyurs-12-en-28-oic acid, 2 alpha, 3 alpha, 19 alpha, 23-tetrahydroxyurs-12-en-28-oic acid, euscaphic acid, beta-sitosterol and daucosterol. The other one was obtained from the acetate of emulsive layer of the petroleum ether and elucidared as 2 alpha, 3 beta-dihydrolup-28-methyl ester diacetate.     

Application of the Salmonella mutagenicity assay and determination of polycyclic aromatic hydrocarbons in workplaces exposed to petroleum pitch and petroleum coke

Workplaces of an Italian carbon electrode factory, exposed to petroleum pitch and petroleum coke, were studied using a coupled chemical and biological approach to evaluate occupational mutagenic/carcinogenic hazards. Analytical procedures for the determination of polycyclic aromatic hydrocarbons (PAH) and Salmonella/microsome mutagenicity tests (with TA98 and TA100 strains) were performed on both industrial ingredients (pitch and coke) and airborne particulate matter of the working environment, after fractionating by sequential Soxhlet extractions with four organic solvents of increasing polarity (benzene, chloroform, methanol and acetone). The results showed: (a) the presence of extraordinarily high PAH (carcinogenic and non-carcinogenic) contents in the benzene extracts of petroleum pitch (3.6 wt% of total PAH) and of airborne particulate samples (up to 0.35 wt% of total PAH), in correlation with very high indirect (after metabolic activation) mutagenic responses of benzene extracts with strain TA98; (b) very high indirect mutagenic responses in the other extracts of the airborne particulate samples (especially with strain TA98); (c) the production during the processing at high temperatures of directly acting mutagens (without metabolic activation) which were absent in the starting materials and their release in the air of workplaces. The comparison of chemical analytical and mutagenicity data has proved to be an interesting approach for better defining the relative health hazards due to occupational exposure to potentially mutagenic/carcinogenic petroleum products.     

The effect of adrenergic agonists on the systemic response to hemorrhage

An in vivo antitumor screening of extracts of Gomphrena martiana indicated positive activity in the petroleum ether extract, and its further bioactivity-directed fractionation resulted in a lipophilic flavonoid fraction. Upon inoculation of various doses of 5,6,7-trisubstituted flavones on two murine tumor lines, Sarcoma 180 and Ehrlich's carcinoma, a decrease of tumor growth was observed. An in vitro KB cultured cell screen indicated cytotoxicity.     

延安地区土壤中石油类污染物测定方法的研究

以三氯甲烷和石油醚的混合溶剂（V三氯甲烷∶V石油醚=2∶1）为提取剂,用重量法和紫外分光光度法测定了延安地区土壤中石油污染物的含量,并对土样中石油类污染物的提取和测量方法进行了比较研究。结果表明,采用单一的有机溶剂如三氯甲烷、石油醚、二氯甲烷、四氯化碳、正己烷和环己烷等作提取剂,石油污染物的提取效果不理想,而采用三氯甲烷和石油醚的混合溶剂（V三氯甲烷∶V石油醚=2∶1）为提取剂,提取效果较好。土壤样品中石油污染物经提取后用紫外分光光度法测定,操作简单、试剂用量少、污染小,结果的精度较好,适合于延安地区土壤中石油类污染物含量的测定。     

Anti-inflammatory activity of seed extracts of Pongamia pinnata in rat

Pongamia pinnata is a marsh growing Indian tree. Its seeds are used in febrile and inflammatory diseases in Indian system of medicine. Previous preliminary studies with ethanolic seed extract of this plant had exhibited an anti-inflammatory effect in rat. Different solvent fractionated extracts were evaluated in the present study for anti-inflammatory effect in chemically induced paw inflammation in rats. Anti-inflammatory effects of P. pinnata were best seen against bradykinin and PGE1-induced inflammation. In contrast minimal effects were seen against histamine and 5-HT-induced inflammation. The predominant action of extracts of Pongamia pinnata appears to be a modulation of eicosanoid-events in inflammation.     

Replacement of chloroform throughout glycosphingolipid isolation

Chloroform, the predominant constituent of solvents used for lipid extraction and chromatography, is believed to give rise to birth defects and serious damage to health, and may also be carcinogenic. Therefore, simple and successful methods have been developed to replace chloroform throughout the isolation of glycosphingolipids (GSLs) by less harmful solvents. Gangliosides of sheep brain (ganglio-series gangliosides GM1, GDla, GD1b and GT1b) and of lymphocyte-derived mouse hybridoma cells (namely GM3) were extracted with six different solvent mixtures. Chloroform:methanol:water (40:80:30, v/v/v) was employed as reference (solvent I). Combinations without chloroform were: n-propanol:water (40:10, v/v) (II), methylisobutylketone:methanol:water (40:80:30, v/v/v) (III), ethylacetate:methanol:water (40:72:28, v/v/v) (IV), methylacetate:methanol:water (40:72:28, v/v/v) (V) and petroleum ether:isopropanol:water (40:112:38, v/v/v) (VI). After extraction and dialysis, the weight of lipid extract as well as the content of sialic acid, gangliosides, sulphatides and phospholipids were determined. Quantitation of GSL yields in crude extracts obtained by the alternative solvent mixtures II to VI showed recoveries of brain gangliosides from nearly 67% up to 104% compared with the reference solvent I. Extraction of hybridoma cells by means of the alternative combinations without chloroform revealed at least the same and mostly better ganglioside yields in the range from 98% to 116% with regard to the reference solvent I. n-Propanol:water (II) and methylisobutylketone:methanol:water (III) were the recommended extractants for both tissues. Therefore, the methods described offer simple, less hazardous and successful strategies for GSL extraction in excellent yield without the need for using chloroform.     

Biotransformation of trithiozine. III - Isolation and identification of further metabolites in rat urine

The metabolism of a new antisecretory and antiulcer drug, trithiozine (I.S.F. 2001, T), was studied in 4 hr rat urine samples after i.p. administration. After extraction at pH 7 with chloroform, the urine was either incubated with beta-glucuronidase or acidified to pH 2 and subsequently extracted with chloroform. The organic layers were evaporated to dryness and the residues used for TLC analysis. The neutral extracts revealed five spots, not present in control rat urine, corresponding to the unchanged drug T and to four metabolites. Two of the metabolites had been previously identified as the 4-(3,4,5-trimethoxybenzoyl)tetrahydro-1,4-oxazine (TBO) and the 4-(3,4,5-trimethoxythiobenzoyl)tetrahydro-1,4-oxazine-S-oxide (TO). Three other metabolites were found in the extracts after beta-glucuronidase incubation. TLC, U.V. and M.S. data were consistent with the structure 4-(3,5-dimethoxy-4-hydroxythiobenzoyl)tetrahydro-1,4-oxazine (HT), the corresponding S-oxide (HO) and the 4-(3,5-dimethoxy-4-hydroxybenzoyl)tetrahydro-1,4-oxazine (HBO). The acidic extracts revealed two spots structurally identified as the 3,4,5-trimethoxybenzoic acid (TBA) and the previous HBO. On the basis of present knowledge, a possible metabolic pathway of T is reported, consisting in a rapid metabolic oxidation on the sulfur atom and a slower demethylation on the para methoxy group. The presence of TBA is indicative of subsequent enzymatic hydrolysis of TBO. The intense and long-lasting inhibitory effect of T on gastric secretion is tentatively correlated with the pharmacological activities of some of its metabolites.     

Contribution of phenolic and quinonoid structures in the mutagenicity of the edible mushroom Agaricus bisporus

The objectives of this work were to establish the contribution of agaritine in the mutagenicity of ethanolic extracts from Agaricus bisporus and to examine the possible involvement of phenolic and quinonoid compounds in the mutagenic response to mushrooms. The mutagenic profile of agaritine in the Ames test, in the absence of an activation system, was different from that of the mushroom ethanolic extracts. Incorporation of rat hepatic cytosolic fractions as the activation system increased the mutagenicity of the mushroom ethanolic extracts in Salmonella typhimurium strain TA 104 but did not influence the mutagenicity of agaritine. It was concluded that agaritine is not the principal mutagenic component in the mushroom. The cytosol-induced mutagenicity of the mushroom extracts required NADPH, and was inhibited by dicoumarol and menadione. Moreover, the mutagenic response in the presence of cytosolic fractions was inhibited by superoxide dismutase, catalase, glutathione and dimethyl sulfoxide, thus implicating reactive oxygen species. Finally, tyrosinase, the enzyme converting mushroom phenols to quinones, increased the mutagenicity of the mushroom extracts. Collectively, the above results indicate that phenolic and quinonoid compounds, presumably through the generation of reactive oxygen species, may play a significant role in the mutagenicity of mushroom extracts.     

CRUD FORMATION IN THE SOLVENT EXTRACTION SYSTEM Zr (IV), HNO3-D2EHPA

The limiting organic concentration (LOC) of zirconium (IV), which is the maximum possible concentration of the metal species in the organic phase without formation of crud as a third phase, was determined under different experimental conditions such as concentration of HNO₃ in the aqueous phase and concentration of a modifier (TBP or isodecanol) in the organic phase. Also, the effect of several diluents (orthoxylene, toluene, n-heptanes, n-dodecan, petroleum ether, chloroform, and kerosene) and the effect of aqueous solution aging on LOC have been investigated. The results showed that the formation of crud depends on the dipole moment of the organic phase, and that aging of the aqueous feed has a significant effect on the LOC of zirconium.     

Studies of the methods used for the determination of the digestibility of crude fats in calves

An investigation was made of methods used for the exact determination of the crude content and crude extract of faeces. The following methods were examined: (1) Following treatment with hydrochloric acid a 1:1 mixture of ether and petroleum ether was used. (2) Following hydrolysis with hydrochloric acid extraction with ether was carried out immediately from the aqueous phases. (3) The analysis was carried out with Folch's solvent. A 1:1 mixture of methanol and chloroform was used. (4) The faeces were dried with infrared lamps and crude fat extraction was carried out with Folch's solvent. A 1:1 mixture of methanol and chloroform was used. (5) The faeces were dried with infrared lamps and extraction with ether was done in a Soxhlet apparatus. Nearly identical results were obtained with methods 1 and 2 or 3 and 4 in determining the content of crude extract, the acid number and the percentage proportion of fatty acids. Essential differences were found between methods 1 and 2 and methods 3 and 4. The most reliable results were obtained by using method (2). Method (5) was found to be most suitable for large-scale testing. Values for the content of crude extract in the faeces were somewhat lower (0.78%-1.2%) when determined by method (5). These variations were not significant statistically. (P greater than 5%). No significant differences were observed between methods (2) and (5). It was thus established that method (5) is best suited for estimating the crude extract of the faces. The faeces of calves were found to contain a high proportion of unsaponifiable matter. This proportion was subtracted from the percentage of crude extract and the amount of faecal crude fat was thus estimated. Very small but statistically significant differences were found by converting digestibility data of crude extract into those of crude fat. Hence, for future work the most appropriate method appears to be the estimation of the crude extract in feedingstuffs.     

Establishment of a protocol for testing antimicrobial activity in southern African macroalgae

Six southern African seaweeds, two Chlorophyta, Phaeophyta and Rhodophyta, were used to test for antibacterial activity against Bacillus subtilis, Staphylococcus aureus, Acinetobacter Iwoffi and Escherichia coli. Extracts were prepared using a range of organic solvents (acetone, chloroform, diethyl ether, 80% ethanol and methanol), by soaking ground seaweed samples in the extraction solvent, evaporating to dryness, and redissolving in the respective solvent. In addition, milled and ground samples were boiled in 80% ethanol for 4 h. Crude extracts were tested for antibacterial activity using three application techniques, viz paper discs, direct spotting and wells, and onto three types of agar plates, viz seeded agar, bacterial lawns and overlay agar. Inhibition zone diameters were measured and analysed by multifactorial analysis of variance. Boiling milled seaweeds in 80% ethanol, with application of the extracts into 6.1 mm wells in overlay agar constituted the test protocol which yielded the highest activity indices.     

Economic reduction of refuse in the hospital

The effects are described of adding either the dried fruiting bodies of the oyster fungus Pleurotus ostreatus, or an ethanolic extract of it, to the diet of normal Wistar male rats and a strain with hereditary hypercholesterolaemia. Addition of the dry oyster fungus to the diet significantly increased, by more than two-fold, the triacylglycerol (TAG) level in the plasma of both groups of rats compared with their respective controls. In contrast, the ethanolic extract did not significantly change TAG levels. Values for total cholesterol and its high- and low-density lipoprotein fractions in the plasma, as well as the calculated atherogenic index, did not show any significant change. Levels of liver cholesterol were significantly lowered by the dried oyster fungus in both hypercholesterolaemic and normal groups of rats, and by the ethanolic extract in normal rats. A significantly increased phospholipid-to-cholesterol ratio in the aortas of both groups of rats, after the administration of either dried oyster fungus or the ethanolic extract of it, suggests a favourable anti-atherogenic effect for both.     

Time courses of hepatic injuries induced by chloroform and by carbon tetrachloride: comparison of biochemical and histopathological changes

The relationship was investigated between biochemical and morphological changes in chloroform (CHCl3)- and carbon tetrachloride (CCl4)-induced liver damage. The time courses of hepatic microsomal cytochrome P450 (CYP) content, hepatic microsomal CYP2E1 activity, hepatic reduced glutathione (GSH) content, plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were examined in relation to the liver morphology in rats orally treated with CHCl3 or CCl4 (3.35 mmol/kg). The CYP content and the activity of CYP2E1 markedly decreased in the CCl4-treated rats 3 h after treatment compared to much lower decreases in the CHCl3-treated rats. The hepatic GSH content was decreased to a similar extent in both groups of rats at 3 h after treatment; in the CCl4-treated rats, the GSH content continued to decrease, reaching a minimum at 24 h and without attaining the normal level at 72 h after treatment. By contrast, hepatic GSH content in the CHCl3-treated rats began to increase from 6 h, attaining complete recovery 48 h after treatment. Plasma ALT and AST activities were significantly elevated by CCl4 as early as 3 h after treatment, while the activities in the CHCl3-treated rats did not increase until 6 h after treatment. In both groups of rats, ALT and AST activities reached a maximum at 24 h, and gradually decreased, remaining at abnormal levels at 72 h. Hepatic cells in the CCl4-treated rats were found to be necrotic as early as 3 h post-treatment, whereas few or no morphological changes appeared in the liver of CHCl3-treated rats. The extent of necrosis was at a maximum 24 h after treatment in both CHCl3- and CCl4-treated rats. In addition, some necrotic cells remained in the liver of CCl4-treated rats 72 h after treatment, while the necrosis in the CHCl3-treated rats was almost negligible. The present results indicate that almost the same time-courses of biochemical and morphological changes were followed in rats of both the CHCl3- and CCl4-treated groups.     

A new diabetes model induced by neonatal alloxan treatment in rats

Anti-leishmanial activity of chloroform and methanol extracts of Vernonia amygdalina, a plant widely used in Ethiopia for the treatment of parasitic infections, has been assessed in vitro on Leishmania aethiopica. Amastigotes were more sensitive to V. amygdalina than promastigotes. The chloroform extract had a stronger parasiticidal activity, with median effective doses (ED50) of 18.5 micrograms/ml and 13.3 micrograms/ml for promastigotes and amastigotes, than the methanol extract with ED50 of 74.4 micrograms/ml and 45.8 micrograms/ml respectively. Cytotoxicity caused by V. amygdalina to host cells, the human leukaemia monocyte THP-1 cell line, as determined by the methyl tetrazolium assay, resulted in a median lethal dose (LD50) of 19.6 micrograms/ml for the chloroform extract and 243.4 micrograms/ml for the methanol extract. In comparison, the ED50 and LD50 of pentamidine, a standard anti-leishmanial drug, were 0.5 micrograms/ml and 1.4 micrograms/ml respectively. These results indicate that V. amygdalina displays potent anti-leishmanial activities and warrants further investigation.     

Liver fibrosis

Surfactant abnormalities may contribute to the impairment of gas exchange observed in Pneumocystis carinii pneumonia. Analysis of rat bronchoalveolar lavage (BAL) lipid extracts from normal controls, steroid controls, trimethaprim-sulfamethoxazole (TMP-SMX) controls, TMP-SMX/P. carinii pneumonia controls, and P. carinii pneumonia animals reveal similar total phospholipid and total protein levels. However, there was a marked reduction in phosphatidylglycerol (PG) from the BAL of P. carinii pneumonia rats as compared with control animals, with a decrease from 4.91 +/- 1.29 nmol/mg protein to 0.46 +/- 0.57 nmol/mg protein (p<0.05) and a decrease, as a percent of total phospholipids, from 7.7% +/- 0.88% to 0.91% +/- 0.59% (p<0.001). Furthermore, in vitro surface activities of BAL lipid extracts from control and P. carinii pneumonia rats revealed minimum surface tension increases from 9.38 +/- 1.71 mN/m in controls to 16.36 +/- 0.83 mN/m in P. carinii pneumonia rats (p<0.05) and likewise maximum surface tension increases from 22.14 +/- 4.34 mN/m to 38.57 +/- 2.07 mN/m (p<0.01). Of interest, the surface activity of PG-deficient P. carinii pneumonia BAL lipid extracts is completely restored to that of normal controls by the addition of exogenous PG. These findings suggest that a functionally abnormal surfactant occurs in P. carinii pneumonia and that this may account, in part, for the impairment of gas exchange observed in this disorder.     

Induction of flat morphology in K-ras-transformed fibroblasts by lycorine, an alkaloid isolated from the tropical plant Eucharis grandiflora

In the course of screening for Ras function inhibitors from plant extracts, we isolated lycorine from a chloroform extract of Eucharis grandiflora leaves. Lycorine induced flat morphology in K-ras-NRK cells after treatment for 2-3 days, whereas its morphological effect on NRK cells was weaker. Lycorine was found to inhibit protein synthesis specifically in cultured K-ras-NRK cells. It also lowered the cellular amount of Ras in 2-3 days.     

Chronic dissection of the thoracic aorta in a patient with tuberculous pleuro-pericarditis

The production of mycotoxins by Alternaria alternata in cellulosic ceiling tiles was examined with thin-layer chromatography and high-performance liquid chromatography procedures. Alternariol and alternariol monomethyl ether were found in ceiling tile extracts, whereas extracts of control rice cultures of all three isolates produced these mycotoxins plus altenuene and altertoxin I. Extensive fungal growth and mycotoxin production occurred in the ceiling tiles at relative humidities of 84-89% and 97%.