The effect of cell excision and microelectrode perforation on membrane resistance measurements of Nitella translucens.

The effect of cell excision and microelectrode perforation on the membrane resistance measurements of Nitella translucens was determined by direct experiment. From the results it is concluded that perforation has no effect on cells as short as 1 cm. Current leakage though the node of an excised cell has however to be given some consideration. The method used for determining the resistance recovery to insertion has a wide application and its simplicity will encourage its use in other biological systems.     

Water Flow Through Vessel Perforation Plates--The Effects of Plate Angle and Thickness for Liriodendron tulipifera

The effects of scalariform perforation plate thickness and anglewithin vessel elements of Liriodendron tulipifera were studiedwith a computational fluid dynamical model. The pressure gradientand hence resistance to flow through the plate increased asthe perforation plate increased in thickness. Increasing theangle of the plate relative to the axis of the vessel (samenumber of pores) also increased the pressure gradient alongthe modelled cell. For the model matching the actual vesselelement, the plate contributed 8% to the flow resistance ofthe vessel element. This contribution increased only to 11%for doubled plate thickness, and to 14% for a plate at an angleof 60° to the vessel axis. The perforation plate alteredthe velocity profile across the vessel element, but to a differentextent depending on the angle of the plate. A plate at an acuteangle to the vessel axis has little effect on the paraboloidprofile as found upstream from the plate, while obtuse angleplates change the flow profile such that fluid through poresnear the wall is accelerated to a greater velocity than foundin the centre of the cell. Key words: Conductance, perforation plate, vessel, water flow     

Microwounding is a pivotal factor for the induction of actin-dependent penetration resistance against fungal attack

Induced penetration resistance is triggered by failed penetration attempts of nonpathogenic fungi. The resistance mechanism is an important nonhost reaction in plants that can block the invasion of filamentous pathogens such as fungi and oomycetes. However, it remains unclear whether the mechanical stimuli accompanying fungal penetration play a role in induced penetration resistance, whereas the perforation of the cell wall may provide significant stimuli to plant cells. Here, we used microneedles or biolistic bombardment to mimic fungal penetration pegs and a micromanipulation transfer technique of the bio-probe, a germling of Blumeria graminis hordei, to the wounded cells to demonstrate that microwounds derived from fungal penetration attempts may trigger induced penetration resistance in plant cells. When preinoculated with the nonpathogenic fungi Erysiphe pisi and Colletotrichum orbiculare, which were unable to penetrate a barley cell, the penetration of a bio-probe that was transferred by micromanipulation onto the same cell was completely blocked. Fungal penetration was essential to the triggering of induced penetration resistance because a penetration-peg-defective mutant of C. orbiculare completely lacked the ability to trigger resistance. The artificial microwounds significantly, but not completely, blocked the penetration of the bio-probe. Treatment with the actin polymerization inhibitor cytochalasin A or expression of the actin depolymerizing protein HvPro1 caused complete ablation of the induced penetration resistance triggered by either failed fungal penetration or artificial microwounds. These results strongly suggest that microwounding may trigger actin-dependent induced penetration resistance. Manipulation of induced penetration resistance may be a promising target to improve basic disease resistance in plants.     

New Mechanisms of Flucytosine Resistance in C. glabrata Unveiled by a Chemogenomics Analysis in S. cerevisiae

5-Flucytosine is currently used as an antifungal drug in combination therapy, but fungal pathogens are rapidly able to develop resistance against this drug, compromising its therapeutic action. The understanding of the underlying resistance mechanisms is crucial to deal with this problem. In this work, the S. cerevisiae deletion mutant collection was screened for increased resistance to flucytosine. Through this chemogenomics analysis, 183 genes were found to confer resistance to this antifungal agent. Consistent with its known effect in DNA, RNA and protein synthesis, the most significant Gene Ontology terms over-represented in the list of 5-flucytosine resistance determinants are related to DNA repair, RNA and protein metabolism. Additional functional classes include carbohydrate and nitrogen—particularly arginine—metabolism, lipid metabolism and cell wall remodeling. Based on the results obtained for S. cerevisiae as a model system, further studies were conducted in the pathogenic yeast Candida glabrata. Arginine supplementation was found to relieve the inhibitory effect exerted by 5-flucytosine in C. glabrata. Lyticase susceptibility was found to increase within the first 30min of 5-flucytosine exposure, suggesting this antifungal drug to act as a cell wall damaging agent. Upon exponential growth resumption in the presence of 5-flucytosine, the cell wall exhibited higher resistance to lyticase, suggesting that cell wall remodeling occurs in response to 5-flucytosine. Additionally, the aquaglyceroporin encoding genes CgFPS1 and CgFPS2, from C. glabrata, were identified as determinants of 5-flucytosine resistance. CgFPS1 and CgFPS2 were found to mediate 5-flucytosine resistance, by decreasing 5-flucytosine accumulation in C. glabrata cells.     

An Induced Resistance Effect of Hydroxyurea on Plants Infected by Meloidogyne javanica

Aqueous solutions of hydroxyurea (HU) in various concentrations were applied as soil drenches to Meloidogyne javanica-infected plants. At a concentration of 15 ppm, the chemical hampered giant cell formation and the number of females on the roots was 20% of that of the control but the growth of the host plants was not affected. Additional HU applications after the one at infection did not add to the inhibitory effect. HU exerted its effect on M. javanica-infected tomato in five soil types. Soil temperature of 32 C neutralized the HU-induced resistance in much the same way that high temperature breaks the natural resistance in M. incognita-resistant tomato. This study provides further evidence of the role of HU as an induced resistance agent.     

Genetics of CHLAMYDOMONAS REINHARDTII Diploids. II. the Effects of Diploidy and Aneuploidy on the Transmission of Non-Mendelian Markers

The transmission of two non-Mendelian drug resistance markers has been studied in crosses of Chlamydomonas reinhardtii involving diploids and aneuploids with different mating type genotypes. Under normal laboratory conditions for gametogenesis, mating and zygote maturation, the transmission pattern of the non-Mendelian markers sr-u-1 (resistance to streptomycin) and spr-u-1-27-3 (resistance to spectinomycin) is primarily determined by the mating type genotypes of the parental cells. Our results confirm and expand an earlier observation suggesting that an apparent codominant function of the female (mt⁺) allele in regulating chloroplast gene transmission in meiosis appears to be distinct and separate from its recessive function in regulating mating behavior. The chloroplast DNA complement (as indexed by the number of extranuclear DNA-containing bodies) may exert a secondary effect on the transmission of these markers. Within a mating type group (mt⁺/mt^- or mt^-/mt^-) a cell line with more chloroplast DNA tended to transmit its non-Mendelian markers more frequently than a cell line with less chloroplast DNA.     

The amiloride resistance gene,car1, of Schizosaccharomyces pombe

Amiloride, an inhibitor of various sodium transporters, is toxic to Schizosaccharomyces pombe at low concentration in minimal but not in rich media. Amiloride-resistant mutants were isolated and shown to represent a new locus (car1 for changed amiloride resistance) on chromosome I. The carl gene was cloned and sequenced. Sequence analysis revealed an open reading frame of 526 amino acids with a predicted molecular weight of 58 545 Da. It has 52% hydrophobic residues and belongs to the class of 12-transmembrane-domain transport proteins. Gene disruption of carl results in increased amiloride resistance. earl has sequence similarity to proteins from Candida associated with resistance to benomyl, methotrexate and cycloheximide. No single physiologically identifiable component of sodium transport appeared to be lost. We propose that earl serves an uptake function, perhaps as a symport with an unknown substrate and this carrier may transport amiloride into the cell. Further, we suggest that amiloride toxicity at low concentrations is not due to its effect on sodium transport but, rather, depends on intracellular interference with an unknown biosynthetic pathway.     

Analysis of the effects of medicinal leech on arterial function in elderly volunteers by means of photoplethysmography with Angioscan-01

The state of the arterial wall in elderly patients after the application of medicinal leech was analyzed using an Angioscan-01 diagnostic device. The effect of one medicinal leech on the endothelial vasomotor function of small resistance arteries and medium-sized muscular arteries was estimated. Arterial wall rigidity was assessed by measuring the arterial stiffness index and the augmentation index during stress testing in the form of brachial artery occlusion. It has been shown that the application of one medicinal leech has not only local, but also system effect on arterial endothelium by improving its vasomotor function through normalization of arterial wall stiffness. This process is supposed to involve the salivary cell secretion of medicinal leech, which is able to raise the NO level both in cells and in extracellular fluid and to activate e-NOS and n-NOS, as it has been shown recently in the culture of human vascular endothelial cells (HUVEC).     

Elimination of the nucleolus from the nucleus of a living cell by centrifugation (a literature review)

The following effects involving the nucleolus take place during centrifugation of living cells at centrifugal forces of several thousand g to several hundred thousand g: settling of the nucleolus in centrifugal direction on the nuclear envelope; pulling the latter as a long stalk with the nucleolus at its end (or alternatively an easy perforation of the nuclear envelope by the nucleolus); release of the nucleolus into the cytoplasm or its expulsion out of the cell; occasional stratification of the nucleolus in the nucleus; fusion of many nucleoli together under centrifugal pressure. The asymmetric topography of the nuclear envelope is considered to be one of the causes of its different resistance to the penetration of the nucleolus. Elimination of the nucleolus from cancer cell nuclei to test the nucleolar contribution to cell malignancy is suggested as one conceivable application of the centrifugal technique of cell enucleolation.     

Effect of interferon alpha on pokeweed mitogen-induced differentiation of human peripheral blood b lymphocytes

In a previous publication, we reported that B-lymphocyte-deprived mice possess a heightened in vivo resistance to a methycholanthrene-induced syngeneic fibrosarcoma. The present in vitro study has disclosed that spleens of such mice possess a significantly higher cytotoxic activity against the same tumor compared to normal rabbit globulin-injected control animals, and that this increase cannot be accounted for by the mere elimination of B lymphocytes. The cell mediating this response was found to be a natural-killer-like cell on the basis of its organ distribution, its full activation without immunization, its target specificity shown directly and by cold target inhibition experiments, and by its surface markers. It is suggested that this increased activity, which does not appear to decline with age, may be responsible for the heightened in vivo antitumor resistance of T mice.     

The mediator of cellular immunity. VIII. Effect of mitomycin C on specifically sensitized lymphocytes.

The antimitotic agent, mitomycin C, was used to probe cellular events underlying the expression of acquired resistance to L. monocytogenes in the rat. It was found that thoracic duct lymphocytes from donors primarily immunized with this organism can protect normal recipients against a Listeria challenge and that their ability to do so is inhibited by treatment of the lymphocytes with mitomycin C. Concentrations of the drug which are effective in this regard also inhibit lymphocyte proliferation, but have little if any effect upon RNA or protein synthesis in cells tested at the time of transfer. Yet mitomycin C probably has irreversible effects on lymphocytes in addition to its capacity to block cell replication, for inhibitor-treated immunoblasts fail to localize in peritoneal exudates induced in recipient rats. The inability of mitomycin-treated immunoblasts to extravasate in inflammatory foci is associated with a diminished capacity of specificially sensitized lymphocytes to promote cell division in peritoneal exudates induced by L. monocytogenes. The results illustrate the importance of lymphocyte-macrophage interactions in cellular resistance to infection and give credence to the view that these interactions occur locally at sites of microbial invasion.     

The expression of plasmid-determined resistance to ampicillin in colicin K-tolerant mutant of Escherichia coli

A large fraction of colicin K-tolerant mutants of Escherichia coli with R28K plasmid expressed the lowered resistance to ampicillin (Lra phenotype). Transduction experiments have shown that in HC-108 mutant the tolPAB and lra mutations are cotransducible. The tolPAB mutation is responsible specifically for the Lra phenotype due to changes in the cell envelope which result in the elimination of some form of barrier whose effect is to restrict the access of substrate to the antibiotic inactivating β-lactamase. The tolPA mutation has no effect on the stability of R28K. Neither does it affect the susceptibility of the cell to chloramphenicol, kanamycin, and streptomycin nor the resistance to these antibiotics determined by R1drd19.     

The inhibition of GHR enhanced cytotoxic effects of etoposide on neuroblastoma

Etoposide, a DNA damage-inducing agent, is widely used to treat neuroblastoma. Etoposide binds to and inhibits topoisomerase II, thereby inducing the DNA damage response. However, the underlying mechanism of etoposide resistance in neuroblastoma remains unclear. The results of the present study revealed that etoposide upregulated growth hormone receptor (GHR) expression levels in etoposide-resistant neuroblastoma cells, suggesting that GHR upregulation may be involved in the underlying mechanism of etoposide resistance. Thus, the combined effect of GHR knockdown and etoposide treatment on cell viability, apoptosis and migration in vitro, as well as tumor growth in mouse xenograft models in vivo, was subsequently analyzed. The results of cell viability and colony formation assays demonstrated that GHR knockdown enhanced the inhibitory effects of etoposide on cell viability and sensitized cells to etoposide. The enhanced cell viability was discovered to be, at least in part, due to the increase in etoposide-induced apoptosis following GHR knockdown. Moreover, the knockdown of GHR enhanced the inhibitory effect of etoposide on cell migration. Mouse xenograft studies confirmed the effects of GHR silencing in etoposide-resistant neuroblastoma progression in vivo. Furthermore, the effects of GHR knockdown in etoposide resistance were hypothesized to occur via the inactivation of the MEK/ERK signaling pathway. In conclusion, the results of the present study provided novel insight into the underlying mechanism of etoposide resistance and a potential target for the treatment of etoposide-resistant neuroblastoma.     

Distribution of scalariform and simple perforation plates within the vessel network in secondary xylem of Araliaceae and its implications for wood evolution

The distribution of scalariform and simple perforation plates along vessels in Arthrophyllum otopyrenum, Meryta tenuifolia, and Polyscias multijuga (Araliaceae) is examined. In all three species, most vessels bear simple perforation plates only, but the combination of simple and scalariform perforation plates in variable ratios also occurs. Aggregated arrangement of scalariform perforation plates along the vessels was statistically confirmed in some vessel portions. The scalariform perforation plates occur mostly in narrow vessels that are grouped in multiples. Within the clade represented by Polyscias and Arthrophyllum, the evolutionary transition from scalariform to simple perforation plates is realized as the gradual elimination of vessels or vessel portions with scalariform perforation plates, but is not accompanied by a gradual decrease of the number of bars per perforation plate. The narrow vessels that are grouped in vessel multiples are likely to retain the ability to develop scalariform plates, which could promote the evolution from simple to scalariform perforation plates as is the case within Meryta.     

THE MECHANISM OF THE INHIBITION OF HEMOLYSIS

The principal conclusion of this investigation is that the inhibitory effect of plasma or serum on hemolysis by saponin and lysins of the same type is similar in nature to the inhibitory effects of certain sugars and electrolytes, which again are similar to the acceleratory effects produced by indol, benzene, and other substances already studied. All these effects, both inhibitory and acceleratory, are the result of reactions between the inhibitors or accelerators and those components of the red cell membrane which are broken down by lysins. The inhibitory effect of plasma on saponin hemolysis has a number of properties in common with the inhibition produced by sugars and electrolytes and with accelerations in general. (a) The temperature coefficient is small and negative. (b) The extent of the inhibition depends on the type of red cell used in the hemolytic system. (c) The most satisfactory measure of the extent of the inhibition, the constant R, is a function of the concentration of lysin in the system, and (d) R is a linear function of the quantity of inhibitor present. It is also shown that the inhibitory effect of plasma, and serum is not entirely dependent on its protein content. The process underlying the phenomenon of lysis and its acceleration or inhibition seems to be one in which the lysin reacts with a component or components of the cell membrane in such a way as to break down its semipermeability to hemoglobin, and in which the accelerator or inhibitor also reacts with the same component in such a way as to increase or decrease the effectiveness of the lysin in producing breakdown. The membrane is considered as being an ultrastructure made up of small areas or spots of varying degrees of resistance to breakdown, the resistances being distributed according to a negatively skew type of frequency curve, and the process of lysis seems to begin with the least resistant spots breaking down first. These spots may be arranged in some regular spatial pattern, and the membrane has also to be regarded as possessing spots of varying rigidity of form. The accelerator or inhibitor changes the resistance of every reactive spot in the ultrastructure by a factor R, which suggests that acceleration and inhibition are results of some over-all effect, such as that of changing the extent to which lysin is concentrated at the surface or partitioned between the material of the membrane and the surrounding fluid. Some kind of combination between the accelerator or inhibitor and the material of the ultrastructure is presumably involved; at first the combination seems to be a loose one and partly reversible, but later some of the loose links are replaced by more permanent combinations involving the same types of bond as are broken down by the lysins themselves.     

Prognostic significance of sealed-off perforation in colon cancer: a prospective cohort study

Background

Perforated colon cancer is a rare complication, but has a high risk of recurrence. However, most studies have not distinguished sealed-off perforation from free perforation, and the prognosis is unclear. The aim of this study was to evaluate the oncologic outcome of colon cancer with sealed-off perforation.

Methods

Eighty-six consecutive patients who underwent resection for colon cancer with sealed-off or free perforation were included. We defined sealed-off perforation as a colon perforation with localized abscess identified on operative, computed tomography, or pathologic findings, with no evidence of free perforation, including fecal contamination and dirty fluid collection in the peritoneal cavity. Oncologic outcomes were compared between patients with colon cancer with sealed-off perforation and free perforation using a log-rank test and Cox regression analysis.

Results

The sealed-off perforation group included 62 patients, and 24 patients were in the free perforation group. TNM stage and lymphatic, venous, and perineural invasion were similar between the groups. The median follow-up period was 28.9?months (range 0–159). The sealed-off perforation group had better prognosis compared with the free perforation group in terms of progression-free survival (PFS) and overall survival (OS), although there were no statistically significant differences in PFS (5-year PFS 53.7% vs. 40.5%, p?=?0.148; 5-year OS 53.6% vs. 22.9%, p?=?0.001). However, in multivariable analysis using the Cox progression test, sealed-off perforation did not show a significant effect on cancer progression (p =?0.138) and OS (p =?0.727).

Conclusions

Colon cancer with sealed-off perforation showed no difference in prognosis compared with free perforation.

     

MiR-138 Acts as a Tumor Suppressor by Targeting EZH2 and Enhances Cisplatin-Induced Apoptosis in Osteosarcoma Cells

Chemotherapeutic insensitivity remains a major obstacle to treating osteosarcoma effectively. Recently, increasing evidence has suggested that microRNAs (miRNAs) are involved in drug resistance. However, the effect of miR-138 on cisplatin chemoresistance in osteosarcoma has not been reported. We used real-time PCR to detect the expression of mature miR-138 in osteosarcoma tissues and cell lines. Cell proliferation, invasion, and migration assays were used to observe changes to the osteosarcoma malignant phenotype. MiR-138 was downregulated in osteosarcoma tissues and cell lines, and miR-138 overexpression negatively regulated osteosarcoma cell proliferation, migration, and invasion. We also verified that EZH2 is a direct target of miR-138. Furthermore, enhancing EZH2 expression reduced the inhibitory effects of miR-138 on osteosarcoma. Proliferation, apoptosis assays and caspase-3 activity assay confirmed that elevated miR-138 expression enhanced osteosarcoma cell chemosensitivity to cisplatin by targeting EZH2. In conclusion, the present study demonstrates that miR-138 acts as a tumor suppressor by enhancing osteosarcoma cell chemosensitivity and supports its potential application for treating osteosarcoma in the future.