Membrane cartridges for endotoxin removal from interferon preparations

The suitability of membrane cartridges for the removal of endotoxin from both distilled water and interferon preparations was examined. The endotoxin concentrations were reduced to 4.0 and 7.3 EU/ml, respectively, when about 4000 ml of distilled water with 20 and 28 EU/ml were passed through the deoxycholate and chitosan immobilized membrane cartridges. When 200 ml of interferon preparation with endotoxin concentration more than 80 EU/ml and pH 3.9 were applied to a deoxycholate immobilized membrane cartridge at a flow-rate of 9 ml/min, the endotoxin concentration was reduced to less than 10 EU/ml. However, if an interferon preparation of 450 ml, with more than 80 EU/ml of endotoxin and pH 3.9 was applied to the chitosan immobilized membrane cartridge at a flow-rate of 18 ml/min, the endotoxin concentration was reduced to less than 10 EU/ml.     

Development of mouse embryos in media containing lectins

Lectins known to stimulate mitosis in cultured cells were evaluated for effects on development of mouse embryos in vitro. Two-cell mouse embryos were cultured in one of the following treatments: Whitten's medium as the control medium; Whitten's medium with 1, 10 or 100 mug/ml concanavalin A; Whitten's medium with 1, 10 or 100 mug/ml leucoagglutinin; Whitten's medium with 1, 10 or 100 mug/ml phytohemagglutinin; Whitten's medium with 1, 10 or 100 mug/ml pokeweed-mitogen; and Whitten's medium with 1, 10 or 100 mug/ml wheat germ agglutinin. Development to the morula stage was blocked in media with 100 mug/ml concanavalin A and 10 and 100 mug/ml wheat germ agglutinin, whereas blastocyst formation was blocked in all pokeweed-mitogen supplemented media. Embryos incubated in 10 and 100 mug/ml wheat germ agglutinin underwent premature cavitation or vacuolation at 24 to 48 h of culture. More embryos formed blastocysts in media with 1 and 100 mug/ml phytohemagglutinin and 10 mug/ml leucoagglutinin than in Whitten's medium (P<0.05). The percentage of embryos hatching was greatest in 1 mug/ml phytohemagglutinin (P<0.05), but it was the same in Whitten's medium, 1 mug/ml concanavalin A and 1 mug/ml leucoagglutinin (P>0.05). Cell division was not stimulated by the lectins; however, it was significantly suppressed in media with 10 and 100 mug/ml concanavalin A, 100 mug/ml phytohemagglutinin, 1, 10 and 100 mug/ml pokeweed-mitogen, and 10 and 100 mug/ml wheat germ agglutinin. Solubility of the zona pellucida in sodium isothicyanate (NaSCN) was reduced in 100 mug/ml phytohemagglutinin, 100 mug/ml leucoagglutinin and 1 mug/ml wheat germ agglutinin media (P<0.05) when compared to Whitten's medium and may have accounted for the reduced hatching observed in these treatments. Development of isolated blastomeres into blastocysts was reduced in media with 1 mug/ml wheat germ agglutinin, 1 mug/ml concanavalin A, and 10 and 100 mug/ml leucoagglutinin (P<0.05) but was similar in media with 1 mug/ml leucoagglutinin and 1, 10 and 100 mug/ml phytohemagglutinin when compared to Whitten's medium (P>0.05). The extent of embryo development in media with lectins depended upon the degree of cytotoxicity and potential biochemical modifications induced in the zona pellucida. Greatest embryo development took place in medium with 1 mug/ml phytohemagglutinin; however, the mechanism was not that of stimulation of cell division or a change in zona pellucida solubility.     

The tumour marker CA 195 in colorectal and pancreatic cancer.

The aim of this study was to measure the serum level of the tumour markers CA 195 and CEA in patients with either colorectal or pancreatic cancer both before and at serial intervals after operation. CA 195 and CEA were measured in 199 patients with colorectal cancer and 52 patients with pancreatic cancer. The median concentrations of CA 195 were 3.0 u/ml (interquartile range 3.0-4.5 u/ml) in patients with a Dukes' stage A lesion, 5.8 u/ml (3.0-18.2 u/ml) in patients with a Dukes' stage B lesion, 6.1 u/ml (3.0-24.7 u/ml) in patients with a Dukes' stage C and 23.8 u/ml (11.1-409.0 u/ml) in patients with metastatic disease (normal range 0-7 u/ml). The median levels of CEA were 2.6 ng/ml (1.7-3.3 ng/ml) for Dukes' stage A, 3.3 ng/ml (1.7-7.2 ng/ml) for Dukes' stage B, 3.7 ng/ml (2.2-7.9 ng/ml) for Dukes' stage C and 34.5 ng/ml (13.3-289.4 ng/ml) for metastatic disease. A rising level of CA 195 or CEA after operation suggested recurrence of the tumour. In none of these patients was the recurrence operable. In patients with pancreatic adenocarcinoma, the level of CA 195 was significantly higher in patients with metastatic disease but it did not discriminate between resectable and unresectable disease. The duration of survival correlated with the initial level of CA 195 (Rs = -0.66, p less than 0.001).     

Inhibitory influence of methotrexate and vincristine on the release of cobalophilins from polymorphonuclear granulocytes

The studies have evaluated the effect of methotrexate and vincristine on the release of cobalophilins (vitamin B12 binding proteins) from resting and functionally stimulated polymorphonuclear granulocytes (PMN). Methotrexate (2.5 micrograms/ml; 5.0 micrograms/ml; 20.0 micrograms/ml; and 50.0 micrograms/ml) and vincristine (0.3 microgram/ml; 0.6 microgram/ml; 2.4 micrograms/ml; and 6.0 micrograms/ml) inhibited the cobalophilins release from resting granulocytes. This effect increased with growing concentrations of these drugs. Stimulated PMN could be shown to release cobalophilins more actively than resting granulocytes. Methotrexate (2.5 micrograms/ml; 5.0 micrograms/ml and 20.0 micrograms/ml) and vincristine (0.3 microgram/ml; 0.6 microgram/ml and 2.4 micrograms/ml) inhibited the phagocytosis-activated release of cobalophilins irrespective of the time of PMN stimulation, i.e. before or after being incubated with latex particles.     

Dextran restores albumin-inhibited surface activity of pulmonary surfactant extract.

We examined the effect of dextran (molecular weight 71,000) in counteracting the surfactant inhibitory action of plasma albumin. The surface adsorption time of 0.5 mg/ml modified natural surfactant (MNS; porcine lung extract consisting of phospholipids and hydrophobic surfactant proteins) with 7.5 mg/ml albumin decreased from 681 to 143 s by addition of dextran at a concentration of 10 mg/ml (P < 0.01). The minimum surface tension of 2.0 mg/ml MNS with 30 mg/ml albumin decreased from over 21 mN/m to below 3 mN/m when dextran was added at a concentration of 10 mg/ml (P < 0.01). Surfactant-deficient newborn rabbits given 10 ml/kg of a liquid containing 2.0 mg/ml MNS with 30 mg/ml albumin had a mean tidal volume 13 ml/kg (P < 0.05). Although the underlying mechanism remains to be elucidated, we conclude that dextran restores the albumin-inhibited surface activity of MNS.     

Enhancement of hatching and trophoblastic outgrowth by mouse embryos cultured in Whitten's medium containing plasmin and plasminogen

Mice were induced to superovulate and 2-cell embryos were cultured in Whitten's medium with 10 mg bovine serum albumin/ml (WM) as control, Medium WM with 2.3, 4.6, 23.1 or 46.2 micrograms plasmin/ml, Medium WM with 14.6, 29.1 or 145.7 micrograms plasminogen/ml, Medium WM with 0.1, 0.2, 1.1 or 2.2 micrograms trypsin/ml; Medium WM with 0.2, 0.3, 1.6 or 3.3 micrograms pronase/ml and Medium WM with 10% heat-treated bovine serum (HTBS). Proteolytic activities in the culture media were evaluated at the start of the culture period and 10 days later. Blastocyst formation was significantly reduced in cultures supplemented with pronase and in the two higher levels of trypsin when compared to that in Medium WM. More embryos developed to the blastocyst stage in Medium WM + 2.3 or 23.1 micrograms plasmin/ml and Medium WM + 14.6 micrograms plasminogen/ml than in Medium WM (P less than 0.05). The incidence of hatching was significantly greater in Medium WM than in all plasminogen- and plasmin-supplemented media except for Medium WM + 29.1 micrograms plasminogen/ml. Although not significantly different, hatching was lower in Medium WM and Medium WM + 0.1 microgram trypsin/ml when compared to Medium WM + HTBS. Similar numbers of embryos completed the hatching process in Media WM, WM + 0.1 or 0.2 micrograms trypsin/ml and WM + 0.3 micrograms pronase/ml. Since dissolution of the zona pellucida occurred within 96 h for embryos cultured in Media WM + 1.6 or 3.3 micrograms pronase/ml and WM + 1.1 or 2.2 micrograms trypsin/ml, hatching could not be evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphological and physiological aspects of digestive processes in the graminivorous primate Theropithecus gelada-a preliminary study

Hindgut fermentation has been suggested to contribute significantly to the digestive process in the gelada (Theropithecus gelada). We therefore hypothesized that in an in vitro fermentation test (Hohenheim gas test, using gas production as measure of microbial digestion) inoculum based on fresh gelada feces would degrade grass to a similar degree as zebra (Equus burchelli chapmani) feces and to a higher degree than that of hamadryas baboons (Papio hamadryas). Additionally, morphology of gelada tongue, salivary glands, stomach, and intestine were examined in this study. Gas production was measured between 4 and 96 hr using animal feces incubated with 200 mg of air-dry hay or mixed concentrate sample. For grass hay, 12-hr gas production was as follows: T. gelada (19.9 ml)>Papio (18.4 ml)>Equus (15.7 ml). After 24 hr, gas production changed: Papio (35.1 ml)>T. gelada (31.9 ml)>Equus (27.9 ml). At 96 hr, Papio was unexpectedly the most effective species with the highest gas production (53.1 ml)>zebra (51.2 ml)>gelada (49.4 ml). With a concentrate standard, 12-hr gas production was as follows: T. gelada (38.5 ml)>Equus (36.8 ml) = Papio (36.4 ml). At 24 hr, gas production differed: Papio (51.7 ml)>Equus (47.0 ml) = T. gelada (46.8 ml). At 96 hr, zebra was the most effective species with the highest gas production (63.9 ml)>Papio (60 ml) = T. gelada (59.9 ml). In conclusion, the results show that the microbial population present in gelada feces is able to ferment forage and concentrate substrates in vitro, although this fermentation did not occur with the expected effectiveness. Future studies should therefore focus also on the bacteria species involved.     

负载金属离子的5A 沸石的体外抗菌实验

目的:探讨负载金属离子的5A沸石的体外抗菌作用。方法:选用金黄色葡萄球菌、铜绿假单孢菌、白色念珠菌,利用倍比稀释法对5A沸石组、磺胺嘧啶银组及载不同浓度Ag+、Zn2+5A沸石共15组进行了体外抗菌试验研究,确定最佳抗菌效果离子负载方案。结果:三种细菌的MIC,双金属离子负载在抗菌上具有协同载Ag+5A沸石分别达到了125μg/ml~500μg/ml、31.25μg/ml~500μg/ml、250μg/ml~500μg/ml;附载Zn2+5A沸石分别达到了12.5mg/ml~25mg/ml、6.25mg/ml~50mg/ml、25mg/ml;负载双离子5A沸石分别为250μg/ml~500μg/ml、62.5μg/ml~500μg/ml、500μg/ml。结论:5A沸石负载金属离子后均具有抗菌作用,且抗菌作用与附载金属离子的量正相关;负载相同质量Ag+的5A沸石较附载Zn2+者具有更强的抗菌作用;双金属离子负载在抗菌上具有协同作用;2%Ag++8%Zn2+与2%Ag++10%Zn2+及4%载银组与阳性对照组无显著性差异。     

Changes in circulating plasma levels of cortisol in lactating and non-lactating dairy cattle during the estrous cycle

Plasma cortisol levels were determined by radioimmunoassay for five lactating cows and five open heifers from blood samples collected daily during the course of a complete estrous cycle. Each animal was fitted with an indwelling jugular catheter to minimize stress from bleeding. Mean plasma cortisol levels were 5.67 ng/ml for the lactating cows and 5.87 ng/ml for the open heifers. Mean values for individuals ranged from 3.79 ng/ml to 6.94 ng/ml in the lactating group, and 3.37 ng/ml to 11.69 ng/ml in the open group. These differences, both between and within groups, were not significant. Mean cortisol values during the estrous cycle ranged from 2.26 +/- 0.93 ng/ml on day one to 9.49 +/- 2.11 ng/ml on day six for the lactating group, and 2.74 +/- 0.52 ng/ml on day six to 14.66 +/- 10.78 ng/ml on day twelve for the open group. Group by day interactions were not significant. Attempts to correlate plasma cortisol with lactation or day of estrus were not significant.     

Circulating proangiogenic cytokines and angiogenesis inhibitor endostatin in untreated patients with chronic lymphocytic leukemia

The serum concentration of two pro-angiogenic cytokines: basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1), and anti-angiogenic factor endostatin in the serum of 80 never treated B-cell chronic lymphocytic leukemia (CLL) patients and 27 healthy volunteers was measured using an enzyme linked immunosorbent assay. The serum levels of both bFGF and TGF-beta1 were found to be significantly higher in the CLL group (median 40.5 pg/ml and 38.6 ng/ml respectively) when compared to the control group (median 9.4 pg/ml and 18.9 ng/ml, respectively) (p<0.001). The levels of endostatin were not significantly different in CLL and control groups (median 12.3 ng/ml and 8.4 ng/ml, respectively) (p=0.09). In the group of CLL patients the level of bFGF was significantly higher in patients with progressive disease as compared with patients with stable disease (median 90.5 pg/ml and 40.5 pg/ml respectively) (p<0.001). Patients in Rai stage III and IV also had significantly higher levels of bFGF than patients in Rai stage 0-II (median 100.1 pg/ml and 29.3 pg/ml respectively) (p<0.001). The levels of both TGF-beta1 and endostatin were lower in patients in Rai stage III and IV (median 28.9 ng/ml and 9.1 ng/ml respectively) than in patients in Rai stage 0-II (42.8 ng/ml and 13.1 ng/ml respectively) (p<0.001 and p=0.002 respectively). The level of endostatin was also lower in the group of CLL patients with progressive disease (median 10.0 ng/ml) as compared to patients with stable disease (median 20.5 ng/ml) (p=0.008). In conclusion, the disturbance in the balance between pro- and anti-angiogenic factors may have an important influence on the course of CLL.     

Sensitivity of beta-hemolytic streptococci to antibiotics]

Susceptibility of 64 beta-hemolytic streptococcal strains isolated from the patients with sore throat was studied by the method of serial dilutions in fluid nutrient medium (Konikov broth). Heterogenecity with respect to the sensitivity was investigated in 34 strains among separate populations of the microbes (10 to 15 in every strain). The MIC of benzylpenicillin, oxacillin and erythromycin ranged within 0.007--0.24 U/ml, 0.02--0.36 gamma/ml and 0.005--0.1 gamma/ml respectively. The MIC of benzylpenicillin with respect to separate populations most sensitive to it was 0.007--0.015 U/ml, while that with respect to the lease sensitive populations ranged from 0.015 to 0.24 U/ml. The respective values for oxacillin were 0.02--0.12 and 0.18--0.36 gamma/ml and those for erythromycin were 0.005--0.025 and 0.05--0.1 gamma/ml. Therefore, the beta-hemolytic streptococci isolated from the patients with sore throat were characterized by a rather high sensitivity to the antibiotics which was important precondition for their efficiency in treatment of the patients with the above disease.     

The Venetian Turpentine Mounting Medium

As a mounting medium to follow aceto-carmine the following modification of Zirkle's is suggested: Venetian turpentine, 25 ml.; phenol, 50 ml.; propionic acid, 35 ml.; acetic acid, 10 ml.; water, 20 ml. The technic can be employed with either root-tip or pollen-mother smears, and has been used with quite a variety of plants. It is especially valuable where it is desired to make temporary mounts permanent. The method is simple, and with reasonable care no displacement of marked cells occurs.     

Proliferation of human embryonic corneal stromal cells in a serum-free medium

Substantial multiplication of stromal cells from human embryonic corneas has been obtained in a basal medium MCDB 104 supplemented with 25 ng EGF/ml, 10 micrograms insulin/ml, 20 micrograms transferrin/ml, 25 ng MSA/ml, 500 micrograms ovalbumin/ml, 50 micrograms LDL/ml, 50 micrograms HDL/ml and 10(-6) M hydrocortisone. Even though the growth rate appears to be similar to that in 10% serum, the cells cease proliferating at a lower density.     

Activities of garlic oil, garlic powder, and their diallyl constituents against Helicobacter pylori

Chronic Helicobacter pylori disease is reduced with Allium vegetable intake. This study was designed to assess the in vivo anti-H. pylori potential of a variety of garlic substances. The garlic materials all showed substantial but widely differing anti-H. pylori effects against all strains and isolates tested. The MICs (range, 8 to 32 microg/ml) and minimum bactericidal concentrations (MBCs) (range, 16 to 32 microg/ml) of undiluted garlic oil (GO) were smaller than those of garlic powder (GP) (MIC range, 250 to 500 microg/ml; MBC range, 250 to 500 microg/ml) but greater than the MIC of allicin (4. 0 microg/ml) (Table 2) present in GP. Allicin (MIC, 6 microg/ml; MBC, 6 microg/ml) was more potent than diallyl disulfide (MIC range, 100 to 200 microg/ml; MBC range, 100 to 200 microg/ml), its corresponding sulfide, but of a strength similar to that of diallyl tetrasulfide (MIC range, 3 to 6 microg/ml; MBC range, 3 to 6 microg/ml). Antimicrobial activity of the diallyl sulfides increased with the number of sulfur atoms. Time course viability studies and microscopy showed dose-dependent anti-H. pylori effects with undiluted GO, GP, allicin, and diallyl trisulfide after a lag phase of ca. 1 to 2 h. Substantial in vitro anti-H. pylori effects of pure GO and GP and their diallyl sulfur components exist, suggesting their potential for in vivo clinical use against H. pylori infections.     

A群脑膜炎球菌多糖菌苗固体总量试验

本试验采用５０℃干烤法在不同实验条件下对Ａ群脑膜炎球菌多糖菌苗的固体总量进行了测定,并对其结果进行了比较。实验结果表明：（１）１ｍ１,２ｍｌ和４ｍ１取样量的固体总量之间无显著性差异。（２）采用２ｍｌ取样量用不同规格称量瓶和不同规格吸管对测定结果无明显影响。（３）采用１ｍ１、２ｍｌ和４ｍｌ取样量试验结果重复性均好。但从标准差及变异系数看,取样２ｍｌ或４ｍｌ者,试验误差更小,结果更为准确。     

负载金属离子的5A沸石的体外抗菌实验

目的：探讨负载金属离子的5A沸石的体外抗菌作用。方法：选用金黄色葡萄球菌、铜绿假单孢菌、白色念珠菌,利用倍比稀释法对5A沸石组、磺胺嘧啶银组及载不同浓度Ag＋、Zn2＋5A沸石共15组进行了体外抗菌试验研究,确定最佳抗菌效果离子负载方案。结果：三种细菌的MIC,双金属离子负载在抗菌上具有协同载Ag＋5A沸石分别达到了125μg/ml~500μg/ml、31.25μg/ml~500μg/ml、250μg/ml~500μg/ml;附载Zn2＋5A沸石分别达到了12.5mg/ml~25mg/ml、6.25mg/ml~50mg/ml、25mg/ml;负载双离子5A沸石分别为250μg/ml~500μg/ml、62.5μg/ml~500μg/ml、500μg/ml。结论：5A沸石负载金属离子后均具有抗菌作用,且抗菌作用与附载金属离子的量正相关;负载相同质量Ag＋的5A沸石较附载Zn2＋者具有更强的抗菌作用;双金属离子负载在抗菌上具有协同作用;2%Ag＋＋8%Zn2＋与2%Ag＋＋10%Zn2＋及4%载银组与阳性对照组无显著性差异。     

Solid-phase extraction procedure to remove organic acids from honey

A solid-phase extraction procedure was applied to remove organic acids from honey. Malic, maleic, citric, succinic and fumaric acids were isolated with an anion-exchange cartridge. The different parameters which affected the extraction procedure were studied and optimised to establish the optimal conditions for maximum recovery of organic acids and minimum extraction of interferences. The optimised procedure used a cartridge which was activated with 10 ml of 0.1 M sodium hydroxide solution (percolation rate 3 ml/min). A 10 ml volume of honey solution was passed at a flow-rate of 0.5 ml/min. The cartridge was washed with 10 ml of water (3 ml/min) and organic acids were eluted with 4 ml of 0.1 M sulfuric acid (0.5 ml/min). This solution was injected directly into the chromatograph. When this procedure was carried out on standard solutions of organic acids, recoveries between 99.2 and 103.4% were found. If this procedure was applied to honey samples these recoveries were also satisfactory and ranged from 62.9 to 99.4%.     

Effect of bis [benzyl N'-(indol-3-ylmethylene)-hydrazinecarbodithioato]-zinc(II) derivatives on wound healing in Sprague Dawley rats

Effects of topical application of Bis[benzyl N'-(indol-3-ylmethylene)-hydrazinecarbodithioato]-zinc(II) (BHCZ) on wound healing and histology of healed wound were assessed. Sprague Dawley rats were experimentally induced wound in the posterior neck area. Tween 20 (0.2 ml of 10%) was applied to rats in Group 1 (negative control). Intrasite gel (0.2 ml) was applied topically to rats in Group 2 as reference. BHCZ at the concentrations 0.2 ml of 25, 50 and 100 mg/ml were applied to Group 3, 4 and 5, respectively. Wound dressed with BHCZ significantly healed earlier than those treated with 10% Tween 20. Also wound dressed with 100 mg/ml BHCZ accelerated the rate of wound healing compared to those dressed with intrasite gel and, 25 mg/ml and 50 mg/ml BHCZ. Histological analysis of healed wound with BHCZ showed comparatively less scar width at wound enclosure and the healed wound contained less macrophages and large amount of collagen with angiogenesis compared to wounds dressed with 10% Tween 20. Results of this study showed that wounds dressed with 100 mg/ml of BHCZ significantly enhanced acceleration of the rate of wound healing enclosure, and histology of healed wounds showed comparatively less macrophages and more collagen with angiogenesis.     

The distribution of blood flow to the reproductive organs of rats near term.

The rate of ovarian and utero-placental blood flow through vessels of less than 25 mum diameter was examined with radioactive microspheres in 5 non-pregnant rats and 19 rats at Day 22 of pregnancy. Total blood flow to the reproductive organs was 0-559 ml/min in the non-pregnant animals and 13-2 ml/min in those near term, a 23-fold difference. The mean ovarian blood flow was high and increased from 0-202 ml/min to 0-845 ml/min. Myometrial and endometrial blood flow increased from 0-156 to 2-24 ml/min. The mean maternal placental blood flow at Day 22 of pregnancy was 0-76 ml/min or 121 ml.,min-1 .100 g-1. Litter size was negatively correlated with mean fetal weight but showed little relationship to mean placental weight or to mean maternal placental blood flow.