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Biomedical subjects

L Qiu

Publications and source records attributed to L Qiu.

At least 19 recordsLinked to original sources

Apoptosis-inducing factor is a major contributor to neuronal loss induced by neonatal cerebral hypoxia-ischemia.

Nine-day-old harlequin (Hq) mice carrying the hypomorphic apoptosis-inducing factor (AIF)(Hq) mutation expressed 60% less AIF, 18% less respiratory chain complex I and 30% less catalase than their wild-type (Wt) littermates. Compared with Wt, the infarct volume after hypoxia-ischemia (HI) was reduced by 53 and 43% in male (YX(Hq)) and female (X(Hq)X(Hq)) mice, respectively (P<0.001). The Hq mutation did not inhibit HI-induced mitochondrial release of cytochrome c or activation of calpain and caspase-3. The broad-spectrum caspase inhibitor quinoline-Val-Asp(OMe)-CH(2)-PH (Q-VD-OPh) decreased the activation of all detectable caspases after HI, both in Wt and Hq mice. Q-VD-OPh reduced the infarct volume equally in Hq and in Wt mice, and the combination of Hq mutation and Q-VD-OPh treatment showed an additive neuroprotective effect. Oxidative stress leading to nitrosylation and lipid peroxidation was more pronounced in ischemic brain areas from Hq than Wt mice. The antioxidant edaravone decreased oxidative stress in damaged brains, more pronounced in the Hq mice, and further reduced brain injury in Hq but not in Wt mice. Thus, two distinct strategies can enhance the neuroprotection conferred by the Hq mutation, antioxidants, presumably compensating for a defect in AIF-dependent redox detoxification, and caspase inhibitors, presumably interrupting a parallel pathway leading to cellular demise.

Amino Acid Chloromethyl Ketones↗

Mutant Bik expression mediated by the enhanced minimal topoisomerase IIalpha promoter selectively suppressed breast tumors in an animal model.

To ensure the success of systemic gene therapy, it is critical to enhance the tumor specificity and activity of the promoter. In the current study, we determined that topoisomerase IIalpha promoter is selectively activated in breast cancer cells. An element containing an inverted CCAAT box (ICB) was shown to be responsible for the breast cancer specificity. When the ICB-harboring topoisomerase IIalpha minimal promoter was linked with an enhancer sequence from the cytomegalovirus immediate early gene promoter (CMV promoter), this composite promoter, CT90, exhibited activity comparable to or higher than the CMV promoter in breast cancer cells in vitro and in vivo, yet expresses much lower activity in normal cell lines and normal organs than the CMV promoter. A CT90-driven construct expressing BikDD, a potent proapoptotic gene, was shown to selectively kill breast cancer cells in vitro, and to suppress mammary tumor development in an animal model of intravenously administrated, liposome-delivered gene therapy. Expression of BikDD was readily detectable in the tumors but not in the normal organs (such as heart) of CT90-BikDD-treated animals. The results indicate that liposomal CT90-BikDD is an effective systemic breast cancer-targeting gene therapy.

Animals↗

Expression of elevated levels of pro-inflammatory cytokines in SARS-CoV-infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS.

The authors have previously shown that acute lung injury (ALI) produces a wide spectrum of pathological processes in patients who die of severe acute respiratory syndrome (SARS) and that the SARS coronavirus (SARS-CoV) nucleoprotein is detectable in the lungs, and other organs and tissues, in these patients. In the present study, immunohistochemistry (IHC) and in situ hybridization (ISH) assays were used to analyse the expression of angiotensin-converting enzyme 2 (ACE2), SARS-CoV spike (S) protein, and some pro-inflammatory cytokines (PICs) including MCP-1, TGF-beta1, TNF-alpha, IL-1beta, and IL-6 in autopsy tissues from four patients who died of SARS. SARS-CoV S protein and its RNA were only detected in ACE2+ cells in the lungs and other organs, indicating that ACE2-expressing cells are the primary targets for SARS-CoV infection in vivo in humans. High levels of PICs were expressed in the SARS-CoV-infected ACE2+ cells, but not in the uninfected cells. These results suggest that cells infected by SARS-CoV produce elevated levels of PICs which may cause immuno-mediated damage to the lungs and other organs, resulting in ALI and, subsequently, multi-organ dysfunction. Therefore application of PIC antagonists may reduce the severity and mortality of SARS.

Acute Disease↗

Steinernema akhursti sp. n. (Nematoda: Steinernematidae) from Yunnan, China.

A new species of entomopathogenic nematode, herein described as Steinernema akhursti sp. n., was recovered from soil samples collected from Yunnan Province, the People's Republic of China. Both morphological and molecular data show congruently that S. akhursti sp. n. belongs to the Steinernema feltiae group. It can be separated from all described Steinernema species by the combined morphological and morphometrical characters of various stages of the nematodes. For the first generation male, the new species can be recognized by spicule length 90 +/- 4.6 microm, spicule tip blunt with an aperture on the ventral side, gubernaculum with a long and needle-shaped cuneus, and tail conoid with a prominent mucron on the tip and a concave on ventral side. For the infective juvenile, the combination of the following characters: body length 812 +/- 19 microm, distance from anterior end to excretory pore 59 +/- 1.5 microm, tail length 73 +/- 2.9 microm, E% 77 +/- 4.5, lateral field with six evenly distributed and identical ridges at the middle body portion, and tail with long and slightly constrict hyaline portion can be used to separate the new species from other nematodes. For the female, the new species is characterized by: tail conoid with a short mucron and slightly swelling anal portion and a symmetrical, slightly protruding vulva with conspicuous double-flapped epiptygma. The nematode can be separated from other described species of Steinernema by DNA sequences of either a partial 28S rDNA or the internal transcribed spacer regions of rDNA and from the closely related species S. feltiae and Steinernema oregonense by cross-breeding tests.

Animals↗

A novel approach to remove nitrogen in a biological aerobic filter (BAF).

Laboratory study on nitrogen removal was carried out using biological aerobic filter (BAF) fed with synthetic domestic wastewater in the temperature range of 20.5-26.5 degrees C in order to gain insight into the mechanism and the influences of operation parameters on nitrite accumulation in the biological nitrogen removal process. Influent loading with 0.26-0.62 kg (m(3)d(-1)) NH4+-N, 0.28-0.63 kg (m(3)d(-1)) TN, 1-2 m h(-1) hydraulic loading and air/ water ratio 3:1, resulted in the removal of NH4+-N and TN by 0.15-0.52 kg (m(3)d(-1)) and 0.18-0.42 kg (m(3)d(-1)) respectively. Hence BAF seems to be a promising nitrogen removal technique. The investigation of nitrite concentrations in the bulk and effluent, the profiles of inorganic nitrogen compounds and the spatial distribution of microbial populations and activity indicated that the nitrite accumulation and shortcut nitrification-denitrification had taken place in the bioreactor. No clear relationships were observed between the nitrite accumulation with the influent loading, temperature and pH value except that nitrite concentrations in the bulk increased with high back washing frequency. The results suggested that the filter structure and operation pattern were the principal factors that cause the nitrite accumulation and shortcut nitrification-denitrification occurring in the biological aerobic filter.

Bacteria↗

Steinernema aciari sp. n. (Nematoda: Steinernematidae), a new entomopathogenic nematode from Guangdong, China.

A new species of entomopathogenic nematode, Steinernema aciari sp. n. was described. It was recovered from a soil sample collected from Haimen town, Shantou district in the eastern coast of Guangdong province, the People's Republic of China during a survey for entomopathogenic nematodes. S. aciari sp. n. belongs to the Steinernema glaseri group. It can be separated from all described Steinernema species by the combined morphological and morphometrical characters of various stages of the nematodes. For male, the new species can be recognized by spicule length (86+/-6.3 microm); spicule tip blunt with a hook-like structure; gubernaculum with a short and Y-shaped cuneus and corpus well-separated posteriorly. For infective juvenile, the combination of the following characters: body length (1113+/-68 microm), distance from anterior end to excretory pore (95+/-3.7 microm), tail length (78+/-5.2 microm), and E % (123+/-7) can be used to differentiate the new species from other nematodes. For female, the tail (conoid with a long mamillate terminus and a distinct postanal swelling) and vulva (slightly protruding from body surface with conspicuous double flapped epiptygma) shapes can be used as diagnostic characters for the new species. The new species can also be distinguished from other Steinernema species by DNA sequences of either a partial 28S rDNA or the internal transcribed spacer regions of rDNA, and from the close related species S. glaseri, Steinernema longicaudum CWL05, and Steinernema guangdongense by cross-breeding test.

Animals↗

Immune biology of macaque lymphocyte populations during mycobacterial infection.

Immune responses of lymphocyte populations during early phases of mycobacterial infection and reinfection have not been well characterized in humans. A non-human primate model of Mycobacterium bovis bacille Calmette-Guerin (BCG) infection was employed to characterize optimally the immune responses of mycobacteria-specific T cells. Primary BCG infection induced biphasic immune responses, characterized by initial lymphocytopenia and subsequent expansion of CD4+, CD8+ and gammadelta T cell populations in the blood, lymph nodes and the pulmonary compartment. The potency of detectable T cell immune responses appears to be influenced by the timing and route of infection as well as challenge doses of BCG organisms. Systemic BCG infection introduced by intravenous challenge induced a dose-dependent expansion of circulating CD4+, CD8+ and gammadelta T cells whereas, in the pulmonary compartment, the systemic infection resulted in a predominant increase in numbers of gammadelta T cells. In contrast, pulmonary exposure to BCG through the bronchial route induced detectable expansions of CD4+, CD8+ and gammadelta T cell populations in only the lung but not in the blood. A rapid recall expansion of these T cell populations was seen in the macaques reinfected intravenously and bronchially with BCG. The expanded alphabeta and gammadelta T cell populations exhibited their antigen specificity for mycobacterial peptides and non-peptide phospholigands, respectively. Finally, the major expansion of T cells was associated with a resolution of active BCG infection and reinfection. The patterns and kinetics of CD4+, CD8+ and gammadelta T cell immune responses during BCG infection might contribute to characterizing immune protection against tuberculosis and testing new tuberculosis vaccines in primates.

Animals↗

Effect of nitric oxide on iron-mediated cytotoxicity in primary cultured renal proximal tubules.

Nitric oxide (NO) has been proved to be a mediator of hypoxic injury in renal proximal tubules (PT), but its effect on iron-induced cytotoxicity has remained little known. In this study, we observed the relationship between NO production and lactate dehydrogenase (LDH) release in primary proximal tubular epithelia co-incubated with different doses of NTA-Fe and lipopolysaccharide (LPS) alone or in combination. NO production was monitored by NO2 concentration in supernatants based on the Griess reaction; while the semi-quantitative RT-PCR was applied to detect the inducible nitric oxide synthase (iNOS) mRNA level induced by NTA-Fe and LPS together. In addition, experimental groups were subjected to reactive oxygen species (ROS) scavengers to determine the impact of the interaction between NO and ROS on iron-mediated cytotoxicity. After a 12-h co-incubation, we found that NTA-Fe increased both LDH release and NO2(-) production in a dose-dependent manner (P < 0.001). The level of iNOS mRNA induced by LPS was enhanced by 500 microM NTA-Fe (P < 0.01), lower or higher concentrations had no effect. However, the supernatantNO2(-) level in the same group did not change significantly (P > 0.05) although tubular injury was aggravated (P < 0.001). The addition of L-arginine increased LDH release from 25.05 +/- 8.36% in the iron group to 38.67 +/- 7.67% in iron plus LPS group (P < 0.05); concomitantly, L-NAME mitigated iron toxicity in LPS-treated PT (P < 0.05). Hydroxyl scavengers provided complete protection against iron-mediated cytotoxicity (P < 0.001), but the decrease of NO2(-) production was only significant in the LPS-treated group. In contrast, SOD was partially effective in the LPS group (P < 0.05) whereas the NO2(-) level in the supernatant was inversely raised (P < 0.05). GSH had no effect on either iron toxicity or NO2(-) production. Thus, we conclude that NO can exacerbate the cytotoxicity caused by NTA-Fe in cultured proximal tubular epithelia, but NO is not the only factor. NTA-Fe could enhance the upregulation of iNOS transcription induced by LPS in a specific concentration range, and its regulation of NO production might also involve a post-transcription mechanism. The hydroxyl group is the major mediator in our model and the pro-oxidant role of NO is probably due to its ability to promote the Fenton reaction and form both ONOO(-) and *OH via its interaction with ROS.

Animals↗

Administration of plasmid DNA expressing human interleukin-6 significantly improves thrombocytopoiesis in irradiated mice.

When people are exposed to large doses of ionizing rays in a short time, hematopoiesis is impaired and hemorrhage is one of the major clinical features. Suddenly decreasing platelet counts are responsible for the life-threatening hemorrhagic complication. Therefore, some cytokines have been used to improve thrombocytopoiesis in various radiation-induced thrombocytopenia models. Current measures for this purpose involve repeated intravenous or subcutaneous injections of recombinant proteins, which are expensive and inconvenient, or gene therapy with viral vectors that could not obviate the risk of infection. We tried to determine the possibility of gene therapy with plasmid vectors for radiation-induced hematopoietic injury, which could overcome the above-mentioned problem. In this study, we describe the enhanced efficiency of radiation on gene transfer with plasmid vector in vivo and the physiological role of expressed human interleukin-6 (hIL-6) in vivo on a radiation-induced thrombocytopenia model. After a single intramuscular injection of plasmid hIL-6 DNA on 6.5-Gy-irradiated mice, the hIL-6 protein level in mouse plasma was determined with enzyme-linked immunosorbent assay (ELISA). The level of hIL-6 began to increase from the 4th day, reached the peak value on about the 11th day, and remained at a higher level on the 28th day. Meanwhile, unirradiated mice injected with the same amount of plasmid DNA showed less hIL-6 on the 11th day after administration. Further experiments demonstrated that the hIL-6 level in 7.5-Gy-irradiated mice was about three times higher than that of 5.0-Gy-irradiated mice, suggesting radiation could improve gene transfer efficiency of plasmid DNA in vivo and might be dependent on radiation doses. The expression of hIL-6 in vivo showed a significant effect on hematopoietic recovery after radiation. Not only the platelet nadir in peripheral blood, but also the number of colony-forming cells in bone marrow rose. The increased platelet counts were partially due to the increase of reticulated platelet that reflected the activity of a given population of megakaryocyte in bone marrow. We conclude that radiation could significantly enhance the gene transfer efficiency of plasmid DNA and that gene therapy with plasmid vectors for radiation-induced hematopoietic injury might be more effective than other diseases without DNA repair.

Animals↗

[Study on changes of serum T helper cell type 1 and 2 cytokines after active immunotherapy in women with unexplained habitual abortion].

OBJECTIVE: To investigate the changes of serum T helper cell (TH)1/TH2 type cytokines after the active immunotherapy in unexplained habitual abortion (UHA) women. METHODS: Concentrations of interleukin (IL)-2, IL-12, interferon (IFN)-gamma, IL-4, IL-10 and transforming growth factor (TGF)-beta 1 were measured by enzyme-linked immunosorbant assay (ELISA) method in sera from thirty-three cases of unexplained habitual abortion (UHA) women before and after active immunotherapy. Thirty normal non-pregnancy (NNP) women and thirty normal pregnancy (NP) women were taken as control. RESULTS: (1) Serum concentrations of IL-2 and IL-12 were higher significantly (P < 0.01) in UHA women [(13.3 +/- 13.8) ng/L and (50.5 +/- 25.8) ng/L, respectively] than those in NNP women [(4.6 +/- 6.4) ng/L and (20.3 +/- 28.2) ng/L, respectively]. Serum concentrations of IL-4 and IL-10 were lower significantly (P < 0.01) in UHA women [(13.8 +/- 1.0) ng/L and (13.5 +/- 0.7) ng/L, respectively] than those in NNP women [(14.5 +/- 1.2) ng/L and (14.9 +/- 2.4) ng/L, respectively]. However, there were no significant differences in serum concentrations of IFN-gamma and TGF-beta 1 between UHA women and NNP women. (2) Serum concentrations of IL-2 were lower significantly (P < 0.05) in NP women [(1.6 +/- 4.3) ng/L] than those in NNP women. Serum concentrations of IL-4 were higher significantly (P < 0.01) in NP women [(16.3 +/- 0.8) ng/L] than those in NNP women. But there were no significant differences in serum concentrations of IFN-gamma, IL-10 and TGF-beta 1 between NP and NNP women. (3) After active immunotherapy, serum concentrations of IL-2 and IL-12 were decreased significantly [(5.6 +/- 9.0) ng/L and (28.5 +/- 40.3) ng/L respectively, P < 0.01] whereas serum concentrations of IL-4 and IL-10 were increased significantly [(14.7 +/- 1.2) ng/L and (15.0 +/- 1.8) ng/L respectively, P < 0.01] in UHA women. In addition, no significant changes in serum concentrations of IFN-gamma and TGF-beta 1 were found in UHA women after immunotherapy. (4) As compared to NNP women. there were no significant differences in serum concentrations of the above-mentioned cytokines in UHA women after active immunotherapy. CONCLUSIONS: The results suggest that profile of TH1/TH2 type cytokines tilts to TH2 immunity in normal pregnancy, but to TH1 immunity in UHA. The active immunotherapy could make a shift from TH1 to TH2 immunity, thus favoring the maintenance of pregnancy.

Abortion, Habitual↗

[Effect of multi-micronutrient on heat adaptation and its probable mechanism].

A multi-micronutrient compound is composed of ascorbic acid, thiamin, riboflavin, vitamin B6, niacin, folic acid, K, Na, Ca, Mg, Cu, Mn, Fe and Zn. In order to explore the effect of the multi-micronutrient on heat adaptation and its probable mechanism, heat-exposed Drosophila melanogaster, mice and rats were observed. The results showed that the multi-micronutrient could significantly increase the survival rate of heat-exposed Drosophila melanogaster and prolong the survival time and swimming time of mice under high temperature. The results indicated that the significant influence of multi-micronutrients on heat adaptation could be the result of enhancing the glucocorticoid receptor activity in hepatic cytosol in the heat-exposed rats, inhibiting liver phospholipase A2 activity in the heat-exposed rats, increasing serum SOD activity and decreasing serum MAD content in the heat-exposed rats, as well as increasing level of HSP70 in the heat-exposed Drosophila melanogaster. The results suggested that the effect of enhancing heat adaptation might be related to improving neuroendocrine regulation, enhancing anti-oxidative function against the damage of free radicals as well as increasing HSP70 content.

Adaptation, Physiological↗

Recognition and ubiquitination of Notch by Itch, a hect-type E3 ubiquitin ligase.

Genetic studies identified Itch, which is a homologous to the E6-associated protein carboxyl terminus (Hect) domain-containing E3 ubiquitin-protein ligase that is disrupted in non-agouti lethal mice or Itchy mice. Itch-deficiency results in abnormal immune responses and constant itching in the skin. Here, Itch was shown to associate with Notch, a protein involved in cell fate decision in many mammalian cell types, including cells in the immune system. Itch binds to the N-terminal portion of the Notch intracellular domain via its WW domains and promotes ubiquitination of Notch through its Hect ubiquitin ligase domain. Thus, Itch may participate in the regulation of immune responses by modifying Notch-mediated signaling.

Animals↗

Neuroprotective activity of tiagabine in a focal embolic model of cerebral ischemia.

Gamma aminobutyric acid (GABA) agonists have been shown to have neuroprotective effects when used after focal or global cerebral ischemia. In this study, we evaluated the neuroprotective effects of a GABA re-uptake inhibitory agent, tiagabine, on focal ischemic brain injury in an embolic model in rats. Tiagabine, injected at 1 h after embolization, significantly reduced brain infarction volume, measured with 2,3,5-triphenyltetrazolium chloride (TTC) histological assay. There were varying degrees of neuroprotective effect exhibited in the other experimental groups however this did not reach significance. These results suggest that tiagabine is neuroprotective when administrated at an early period after the ischemic brain injury.

Animals↗

Monomeric midkine induces tumor cell proliferation in the absence of cell-surface proteoglycan binding.

Midkine (MK), a retinoic acid-inducible heparin-binding protein, is a mitogen which initiates a cascade of intracellular protein tyrosine phosphorylation mediated by the JAK/STAT pathway after binding to its high affinity p200(+)/MKR cell surface receptor in the G401 cell line [Ratovitski, E. A. (1998) J. Biol. Chem. 273, 3654-3660]. In this study, we determined the biophysical characteristics of purified recombinant murine MK and analyzed the requirements for ligand multimerization and cell surface proteoglycan binding for the G401 cell mitogenic activity of MK. Our studies indicate that the secreted form of MK (M = 13 kDa) exists in solution as an asymmetric monomer with a frictional coefficient of 1. 48 and a Stokes radius of 23.7 A. By constructing bead models of MK using the program AtoB and the program HYDRO to predict the hydrodynamic properties of each model, our data suggest that MK has a dumb-bell shape in solution composed of independent N- and C-terminal domains separated by an extended linker. This asymmetric MK monomer is a biologically active ligand with mitogenic activity on G401 cells in vitro. Neither heparin-induced formation of noncovalent MK multimers nor tissue transglutaminase II covalent multimerization of MK enhanced MK mitogenic activity in this system. Since neither heparin competition nor cell treatment with chondroitinase ABC or heparinase III abolished the mitogenic effects of MK on G401 cells, cell-surface proteoglycan binding by MK does not appear to be a requirement for its observed mitogenic effects. These results provide strong evidence that the MK-specific p200(+)/MKR has distinctive biochemical properties which distinguish it from the receptor tyrosine phosphatase cell-surface proteoglycan PTPzeta/RPTPbeta and support the hypothesis that the diverse biological effects of MK are mediated by multiple cell-specific signal transduction receptors.

Animals↗

Using deuterium as an isotopic tracer to study the energy metabolism of infective juveniles of Steinernema carpocapsae under aerobic conditions.

Changes in survival, mean dry weight, levels of key energy reserve compounds and respiration of non-feeding infective juveniles (IJs) of Steinernema carpocapsae incubated in various ratios of D2O/H2O on a shaker at 28 degrees C were determined. Patterns of deuterium distribution in trehalose, glycogen and key fatty acids of the IJs incubated in 50% (v/v) D2O/H2O were also examined. The rates of decline in mean dry weight and lipid levels of IJs incubated in D2O/H2O were proportional, while the survival times of IJs were inversely proportional, to the ratio of D2O/H2O. Deuterium was randomly and extensively incorporated into the C-H bonds of trehalose and glycogen but was barely incorporated in fatty acid moieties of the IJs. The changes in the patterns and the extents of deuterium incorporation, as well as the levels of trehalose, glycogens and fatty acids during the experimental period indicate that: (1) The lipogenesis pathway is not functioning in the Ijs. (2) Trehalose and glycogen are constantly consumed and replenished and they are mainly derived from lipids. (3) Futile cycles involving trehalose and glycogen, which enable IJs to regulate the levels of these two compounds more effectively, may exist. The results support the view that lipids are the primary energy reserve of the IJs while trehalose, glycogen and proteins can be used for energy generation, even though this is not their primary role.

Aerobiosis↗

Permeability of the infective juveniles of Steinernema carpocapsae to glycerol during osmotic dehydration and its effect on biochemical adaptation and energy metabolism.

Permeability of the sheath and cuticle of the infective juveniles (IJs) of Steinernema carpocapsae to glycerol and its effect on biochemical adaptation of the IJs to osmotic dehydration were examined by incubating both sheathed and exsheathed IJs in glycerol-d5 solution then monitoring the changes in levels of deuterium labelled and non-labelled glycerol and trehalose. Energy metabolism of the IJs during osmotic dehydration and subsequent rehydration and the effect of the permeated glycerol on this process were investigated by examining and comparing the changes in mean dry weight and key biochemical composition of the IJs dehydrated in glycerol and sodium chloride solutions. The results show: (1) similarly to evaporative dehydration, osmotic dehydration induces IJs to synthesise the protectants glycerol and trehalose; (2) glycerol permeates the sheath and the cuticle into the body of IJs during dehydration in glycerol solution. Part of the permeated glycerol plays a role as protectant like that synthesised by IJs from their energy reserve materials while part is incorporated into trehalose; (3) the sheath reduces the rate of permeation of glycerol and therefore affects the equilibrium glycerol and trehalose levels of the IJs and also the time needed to reach the equilibrium levels; (4) the reduction in mean dry weight and lipids of the IJs during dehydration in glycerol solution is substantially less than those dehydrated in sodium chloride solution. Both the total protectant level and the ratio of glycerol to trehalose of the IJs dehydrated in glycerol solution are higher than those dehydrated in sodium chloride solution; (5) glycogen reserves of the IJs play a role as a buffer reservoir of the protectants during both dehydration and rehydration but the principal sources of the protectants during dehydration are more likely to be lipids and proteins rather than glycogen.

Adaptation, Physiological↗

Inhibition of melanin synthesis by cystamine in human melanoma cells.

In studies to determine whether pigmentation can be regulated physiologically by thiols, human melanoma cells (MM418c5) and melanocytes were found to become depigmented when cultured continuously in 50 microM cystamine. Cystamine was depleted from the culture medium and the treatment was nontoxic and reversible. Cysteamine, dithiothreitol, and phenylthiourea were less effective, and glutathione, cysteine, and cystine were inactive. Tyrosinase (dopa oxidase) activity was not greatly affected except for induction of a lag period. In contrast, tyrosinase activity in an amelanotic melanoma cell line (MM96L) was rapidly inhibited without consumption of cystamine/cysteamine, in association with the generation of free thiol in the culture medium, and could be enhanced by the cystine transport inhibitor, glutamate. Tyrosinase expressed by a recombinant vaccinia virus was inhibited by cystamine treatment of MM96L and HeLa cells. Cystamine treatment lowered the degree of cross-linking of the pigmentation antigen gp75/TRP-1 in MM418c5 cells. Tyrosinase protein and mRNA levels in MM418c5 cells were not affected by cystamine. The results show that cystamine at a concentration close to physiologic levels has multiple effects on the melanogenic pathway. In amelanotic cells, tyrosinase has a short half-life and is readily inhibited by cystamine/cysteamine whereas tyrosinase in the more mature melanosomes of the pigmented cell appears to be less accessible to proteolytic and thiol attack. Inhibition of melanin synthesis in the latter cell type may arise to a significant degree from reduction of cystamine to cysteamine, which sequesters quinones.

Cystamine↗

Histone deacetylase inhibitors trigger a G2 checkpoint in normal cells that is defective in tumor cells.

Important aspects of cell cycle regulation are the checkpoints, which respond to a variety of cellular stresses to inhibit cell cycle progression and act as protective mechanisms to ensure genomic integrity. An increasing number of tumor suppressors are being demonstrated to have roles in checkpoint mechanisms, implying that checkpoint dysfunction is likely to be a common feature of cancers. Here we report that histone deacetylase inhibitors, in particular azelaic bishydroxamic acid, triggers a G2 phase cell cycle checkpoint response in normal human cells, and this checkpoint is defective in a range of tumor cell lines. Loss of this G2 checkpoint results in the tumor cells undergoing an aberrant mitosis resulting in fractured multinuclei and micronuclei and eventually cell death. This histone deacetylase inhibitor-sensitive checkpoint appears to be distinct from G2/M checkpoints activated by genotoxins and microtubule poisons and may be the human homologue of a yeast G2 checkpoint, which responds to aberrant histone acetylation states. Azelaic bishydroxamic acid may represent a new class of anticancer drugs with selective toxicity based on its ability to target a dysfunctional checkpoint mechanism in tumor cells.

Cell Cycle↗