Variation In Surface Proteins In Tetrahymena*

SYNOPSIS. Surface proteins of Tetrahymena were identified by lactoperoxidase iodination, and comparisons were made between a number of strains and species within the genus. an adequate procedure for strain comparisons was found to be solubilization of whole cells following iodination, separation of total cell protein using polyacrylamide gel electrophoresis, and identification of surface proteins by autoradiography of dried gels. the results obtained in the present study show the existence of both interspecific and intraspecific variation in surface proteins of Tetrahymena, but the differences tend to be small within species and large between species. the relation of these cell surface fingerprints to the present taxonomic designations within the genus is discussed. Questions are raised about the functional significance of these surface proteins.     

C-Myc Expression In Exfoliated Cells In Serous Effusions

In an attempt to improve the discrimination between benign and malignant cells in pleural and peritoneal fluids, 32 effusions were investigated for c-myc expression. Smears were prepared from cells harvested from the fluids for immunocytochemical staining to identify the presence of c-myc protein. Recombinant DNA technology (Northern blotting, slot blotting and in situ hybridization) was used to detect c-myc mRNA. No significant difference in expression of c-myc was noted in benign or malignant effusions. Although the results are inconclusive, the recombinant DNA technology developed for this research could be used to investigate the expression of other oncogenes in cytological material.     

In memoriam

Microcosm experiments were performed to identify the influence of bacterial cell surfaces on the morphology, mineralogy, size and solubility of CaCO₃ precipitated in response to the enzymatic hydrolysis of urea in an artificial groundwater (AGW) by the ureolytic bacteria, Bacillus pasteurii. In each microcosm, B. pasteurii were contained within a cellulose dialysis membrane (10 K Dalton MWCO), resulting in bacteria-inclusive and bacteria-free AGW solution. Urea hydrolysis by B. pasteurii resulted in the production of ammonium and an increase in pH in the whole AGW solution. This initiated predominantly rhombohedral calcite precipitation at the same critical saturation state ( S _critical = 12) in the B. pasteurii-inclusive and bacteria-free zone of the AGW, indicating the mineralogy and morphology of CaCO₃ precipitation is not controlled by B. pasteurii surfaces. However, the temporal evolution of distinctly different lognormal crystal-size-distributions in the B. pasteurii-inclusive and bacteria-free zone of the AGW resulted from identical changes in bulk solution chemistry. Specifically, B. pasteurii increased the size and size variance of crystals, and led to a greater crystal growth rate throughout the experiments, relative to bacteria-free AGW. Calculated crystal solubility (ln K _S0 ) was lower for crystals > 4000 nm in diameter, reflecting smaller molar surface areas. This suggests that the larger crystals generated in the presence of B. pasteurii have a lower affinity for re-dissolution than those generated in the bacteria-free AGW, which may act as a positive feedback to maintain larger crystal sizes in the presence of B. pasteurii. During ureolysis, higher bacterial concentrations may therefore generate larger and less soluble carbonate crystals. This has important implications for the adaptation of bacterial ureolysis as a method for precipitating calcium carbonate and co-precipitating metals and radionuclides in contaminated aquifers.     

In Appreciation

Marine animals can induce metallothioneins (MTs) in their responses to exposure to certain trace metals in the environment. MTs generally function as metal storage/detoxification or homeostatic regulation of both essential and non-essential metals. This review discusses the important roles of MTs in metal biokinetics other than metal detoxification and homeostasis in marine animals. Recent studies have revealed the complicated cellular and biochemical processes involving intracellular ligands (cytosolic proteins and insoluble deposits) during metal uptake and elimination. The responses of metal biokinetics to MT induction are metal- and organism-specific. Depending on the different marine animals and metals, all biokinetic parameters such as dissolved metal uptake rate, dietary assimilation efficiency and elimination (efflux) rate can be significantly impacted by MT induction. Among the different metal biokinetic parameters, dietary assimilation efficiency and elimination rate appear to be most impacted by MT induction. MT turnover kinetics can also significantly affect metal uptake kinetics, but again, such influence is also dependent on the organism, particularly its predominant pathway of metal detoxification. Even though the total MT pool in aquatic animals may remain constant, the turnover of MTs, involving MT synthesis and breakdown, can potentially lead to a major change of metal accumulation biokinetics. We propose several issues that need to be further addressed in studying the interaction between MT induction and metal accumulation biokinetics.     

In Vitro and In Vivo Characterization of Pyocin

Pyocin, a bacteriocin obtained from lysates of ultraviolet-induced cultures of Pseudomonas aeruginosa was characterized in vitro and in vivo after 1,000-fold purification by chemical, column, and differential centrifugation procedures. Electron micrographs of negatively stained pyocin preparations contained rod-shaped particles which resembled the contractile tail protein of the T-even phages of Escherichia coli. Although two separate and distinct pyocin fractions were eluted from diethylaminoethyl cellulose (pH 7.5) during the purification procedure, the particles appeared identical. In addition, the two fractions exhibited a close correlation between their titers and the particle numbers as observed in the electron microscope. The particles were approximately 20 by 90 mmu with a core diameter of 5 mmu and a sheath length of 50 mmu. Neither intact phage nor ghosts were seen in any of the preparations, although ringlets of two different diameters, which appeared to correspond to the diameters of the sheath and inner core, were observed. Other studies indicated that, although crude preparations were stable to freezing and thawing, purified preparations lost all of their activity under similar treatment. However, the addition of 50% glycerol to purified preparations completely protected activity. Conversely, aged normal human or rabbit sera enhanced the antibacterial activity of pyocin approximately fourfold, although serum albumin and hemoglobin had no effect. In vivo studies indicated that purified pyocin was not lethal for mice when injected intraperitoneally in concentrations of 28,000 to 1,400,000 units (5.6 to 276 mug of protein), nor was 7,200 to 36,000 units dermonecrotic for rabbits.     

In Memoriam

Practicing physicians are just becoming aware of the animal rights movement, which during the 1980s spawned numerous acts of violence against research facilities throughout the United States. The animal rightists are challenging physicians to show moral justification for the human exploitation of nature and the world of subhuman species. They have aroused public interest in animal welfare, sparked protective legislation for experimental animals, and indirectly encouraged the creation of committees to oversee the conduct of animal experimentation and the conditions of animal confinement. This controversy has necessitated a closer look at the questions of animal experimentation and animal rights against the backdrop of human experimentation and human rights. Physicians and specialists in animal care seek to alleviate suffering and anxiety, and, as moderates, they may be able to bring both sides of the animal rights controversy together in a spirit of mutual tolerance and in the common cause of promoting both human and animal welfare.     

In Memoriam

The Patient Self-Determination Act was implemented in December 1991. Before and after its implementation, we used a structured interview of 302 randomly selected patients to determine their awareness, understanding, and use of advance directives. Implementation of the Act did not have a major effect on these. Although more than 90% of patients were aware of the living will, only about a third selected the correct definition or the correct circumstances in which it applied, and less than 20% of patients had completed one. About a third of patients were aware of a Durable Power of Attorney for Health Care and chose the correct definition, and about half identified the correct circumstances in which it applies; less than 10% had completed such a document. Surprisingly, patients who said they had completed advance directives did not demonstrate better understanding of these documents. Our results indicate that many patients, including some who have completed advance directives, do not fully understand them. It may be unwise to regard these documents as carefully considered, compelling statements of patients'' preferences. Appropriate responses to our findings include increased public education, revising state statutes to bring them into congruence with public perception, and expanding the dialogue between physicians and patients.     

In Memoriam

A key element of the self‐critique in contemporary anthropology centres on the idea that categories such as culture and society have exaggerated human differences. This paper argues that the work of Anthony Cohen, which has attempted to frame “localities” as a new anthropological object, can be criticized on similar grounds. A close reading of Cohen's theoretical and ethnographic writings shows that the “anthropology of locality”, while claiming to be a critique of nationalhomogenization, in fact replicates ata lower level of scale features of the national units. Consideration of this particular example of the exaggeration of (local) difference, moreover, occasions reflection on the wider critique of generalization within anthropology.     

肝硬化疾病与支链氨基酸应用研究进展

蛋白质-营养不良是肝硬化病人最常见的并发症之一。肝脏作为蛋白质、脂类和糖代谢的主要器官,病变后的代谢紊乱随之而来。不适宜的蛋白质-能量摄入只会加重病情最后发生肝性脑病等危及生命的严重后果。因此,肝硬化病人的营养管理显得尤为重要,氨基酸的适宜供给无疑是营养治疗的重中之重。已知支链氨基酸能通过刺激肝细胞合成、减少肝损伤后的分解代谢等诸多方式改善营养状况,但是各种试验结果仍存在争议。最佳适宜量究竟多大,安全性范围的设定以及确切的保护机理等问题仍待进一步深入研究。     

Antithrombotic Activities of Luteolin In Vitro and In Vivo

The objectives of present study were to investigate whether luteolin affects procoagulant proteinase activity and fibrin clot formation and influences thrombosis and coagulation in Sprague–Dawle rats. Luteolin significantly inhibited the enzymatic activity of thrombin and FXa activity by 29.1% and 16.2%. Luteolin also inhibited fibrin polymer formation in turbidity and microscopic analysis using fluorescent conjugate. Coagulation assay of luteolin was found to prolong activated partial thromboplastin time and prothrombin time. Moreover, luteolin protected the development of oxidative stress induced thrombosis in the FeCl₃‐induced carotid arterial thrombus model. This study demonstrated that luteolin may be useful by reducing or preventing thrombotic challenge and can help us better understand the antithrombotic action of luteolin.     

In Vivo,In Vitro,and In Silico Characterization of Peptoids as Antimicrobial Agents

Bacterial resistance to conventional antibiotics is a global threat that has spurred the development of antimicrobial peptides (AMPs) and their mimetics as novel anti-infective agents. While the bioavailability of AMPs is often reduced due to protease activity, the non-natural structure of AMP mimetics renders them robust to proteolytic degradation, thus offering a distinct advantage for their clinical application. We explore the therapeutic potential of N-substituted glycines, or peptoids, as AMP mimics using a multi-faceted approach that includes in silico, in vitro, and in vivo techniques. We report a new QSAR model that we developed based on 27 diverse peptoid sequences, which accurately correlates antimicrobial peptoid structure with antimicrobial activity. We have identified a number of peptoids that have potent, broad-spectrum in vitro activity against multi-drug resistant bacterial strains. Lastly, using a murine model of invasive S. aureus infection, we demonstrate that one of the best candidate peptoids at 4 mg/kg significantly reduces with a two-log order the bacterial counts compared with saline-treated controls. Taken together, our results demonstrate the promising therapeutic potential of peptoids as antimicrobial agents.     

Circadian Variation In Migration Velocity In Small Intestinal Epithelium

Abstract. The variation in migration rates of cells within the small intestinal epithelium was studied over a 24-hr period at 3-hr intervals (migration of cells was studied independently for the crypts and the villi using the changing distributions of [³H]TdR labelled cells as an indicator of cell migration).
Clear changes in the rates of cell movement were observed during a 24-hr period for both crypt and villus epithelium. the rates of cell migration in these two compartments did not correlate well with the exception of samples taken at 18.00 hours. At this time of day there appeared to be no cell movement at all in either crypts or villi. There was not a good correlation between the migration velocity throughout the day and the changes in the number of mitoses.
It is proposed that mitotic rates do not directly govern migration rates but that the converse may be true. Further, the lack of correlation between crypt and villus migration rates at any time of day suggest that the mechanisms controlling all movement in these two regions of small intestinal epithelium may be different.     

In Memoriam

Thirteen cows, Experiment I, were injected at 1 h post-partum with saline (n=4) or Human Chorionic Gonadotropin, (HCG) (n=9). Eleven additional cows, Experiment II, were ovariectomized (ovx) unilaterally at 10 days pre-partum (n=4) or 4 h post-partum (n=7) to remove the only corpus luteum (CL). These ovx post-partum animals (n=7) were also injected as part of Experiment I with saline (n=2) or HCG (n=5). All CL were sliced and incubated in media alone and with LH, tritium-labeled-pregnenolone (³HP₅) or both. Blood samples were collected pre- and post-injection, Experiment I and pre- and post-surgery, Experiment II. Plasma and media samples were analyzed for progesterone (P₄) by radioimmunoassay. Media samples with ³HP₅ were chromatographed for determination of P₅ conversion to P₄. No significant differences (P > 0.1) between P₄ profiles for saline or HCG treated animals were observed. Pre- and post-partum CL slices were able to convert P₅ to P₄. Post-partum CL slices were unresponsive to LH whereas pre-partum slices responded. These data indicate the decline in luteal function peri-partum is probably not due to withdrawal of gonadotropin support.     

Regulation of Pyruvate Decarboxylase In Vitro and In Vivo

Results presented in this paper strongly support the view thatregulation of the key enzyme of alcoholic fermentation, pyruvatedecarboxylase (PDC), is achieved in a number of ways, all associatedwith possible lowering of the cytoplasmic pH during anoxia.These mechanisms include not only the well-known acid pH optimumof PDC, but also long-term, reversible changes in characteristicsof the enzyme established both in vitro and in vivo. Following transfer of desalted extracts from pH 6.0 to 7.4,maximal activity of PDC was decreased, while there was a considerableincrease in the lag before maximal activity was reached. Similarchanges in enzyme characteristics were observed when wheat (Triticumaestivum L. cv. Gamenya) roots and rice (Oryza sativa L. cv.Calrose) coleoptiles were transferred from anoxic to aerobicsolutions, provided PDC was assayed within 10 min of the startof maceration. All of the above changes were usually readilyreversible when extracts were returned to pH 6.0, or when plantswere returned to anoxic solutions. Additional regulation of PDC would be achieved by the S_0.5 forpyruvate which is 0.75 mol m^–3 at pH 6.0, 1.0 mol m^–3at pH 6.8, and 2.5 mol m^–3 at pH 7.4; the latter is wellabove estimates for pyruvate concentrations in the cytoplasmof aerated tissues. We assess that the combined effects of the acid pH optimum,the high S_0.5 at pH 7.4 and the long-term decreases in activityobserved during incubation at pH 7.4 would reduce PDC activityin aerobic cells to at most 7% of the activity in anoxic cells.Possible additional controls for the pathway of alcoholic fermentationare briefly considered. Key words: PDC, regulation, anoxia     

Changes In Stem Cell Compartments In Mice After Hydroxyurea

Hydroxyurea injection kills only approximately 10% of CFU, which are in the S phase of the cell cycle. In mice given a single injection of hydroxyurea CFU_s in the femur decreased only about 50% in 2–4 days after hydroxyurea, and then started to return to normal levels with an overshoot evident after the eighth day after hydroxyurea. CFU_s in spleens of mice given a single injection of hydroxyurea show an evident overshoot as early as 3 days after the drug, reaching levels which are about 300% of normal. CFU_s from blood disappear rapidly, but equally rapidly return to normal values followed by a significant overshoot. the radioresistance of mice increased up to 4 days after hydroxyurea.     

Custom In The Courtroom, Law In The Village: Legal Transformations In Papua New Guinea

After more than twenty-five years on the legal landscape of Papua New Guinea, 'customary law' is ripe for reassessment, particularly as it appears to be an ideal mechanism with which the Papua New Guinean state can meet some of its obligations to a burgeoning body of international law. This article addresses the need to understand customary law in the context of its varying usage across different legal domains in an archetypally pluralistic state. In contrast to older approaches focusing on the problematic interface between an exogenous legal system and indigenous methods of dispute settlement, my concern is with the ways in which these distinct legal forms have fared in each other's company since independence in 1975. Case-studies from a village court and an urban national court demonstrate that village court magistrates and high court judges alike use custom and law as strategic sources of authority. While village courts take custom for granted and therefore must 'discover' law, high courts take law for granted and must 'discover' custom. These processes indicate that, rather than being hybridized as 'customary law', the distinctiveness of custom and law are often maintained in order for one to appear as a resource upon which the other can draw.     

Selective Pressures on RNA Hairpins In Vivo and In Vitro

Comparison of the most stable potential hairpins in the sequences of natural ribozymes with those in the randomized sequences has revealed that the hairpin loop energies are lower than expected by chance. Although these hairpins are not necessarily parts of functional structures, there is a selective pressure to diminish the destabilizing free energies of the hairpin loops. In contrast, no significant bias is observed in the stacking values of the most stable stems. In the ribozymes isolated in vitro the loops of potential hairpins are closer to random values, which can result in less efficient folding rates. Furthermore, the effects of kinetic traps seem to be more significant in the folding pathways of the in vitro isolates due to a potential to form stable stacks incompatible with the functional folds. Similarly to natural ribozyme sequences, the untranslated regions of viral RNAs also form hairpins with relatively low loop free energies. These evolutionary trends suggest ways for efficient engineering of improved RNA constructs on the basis of analysis of in vitro isolates and approaches for the search of regions coding for functional RNA structures in large genome sequences. Received: 12 January 2001 / Accepted: 21 May 2001