Biomedical subjects

W Li

Publications and source records attributed to W Li.

At least 523 records · Page 29Linked to original sources

PubMed1999

Experimental analysis of flow calculations based on HRCT imaging of individual bifurcations.

Flow simulations in airways and arteries allow the non-invasive study of transport and deposition processes in these vessel systems. Individual vessel geometries as input for such simulations are highly desirable. Computed tomography (CT) permits the acquisition of binary data to reconstruct such geometries. To prove the suitability of this reconstruction method, we compared measured with simulated velocities within model bifurcations. Particle image velocimetry was applied to measure flow velocities. Numerical simulations of these velocities were carried out by using the CT data of the same models as input to flow calculations (CFD). Within the resolution limits good agreement between measured and simulated velocities was found. For the smallest bifurcation (tube diameter: 2 mm) the agreement was less, indicating a methodical limitation by the actual resolution of the CT-scan technique. The study showed that a combination of CT and CFD can be considered as an appropriate step towards realistic simulations of particle transportation and deposition in individual geometries of the respiratory or cardiovascular systems.

Algorithms↗

PubMed1999

Contextual effects of transforming growth factor beta on the tumorigenicity of human colon carcinoma cells.

Transforming growth factor betas (TGF-betas) are a growth factor family with negative autocrine growth functions for most epithelial cells including colon carcinoma cell lines. Both type I (RI) and type II (RII) transmembrane TGF-beta receptors have been shown to be indispensable for TGF-beta-mediated cell growth regulation. Previous studies using different model systems have shown that both overexpression of TGF-beta1 and transfection of antisense TGF-beta1 to reduce TGF-beta1 expression could lead to increased tumorigenicity. These results are seemingly contradictory and suggest that effects of TGF-beta modulation on malignant properties of cancer cells may be contextual. This study addresses this issue using human colon carcinoma cells (CBS and FET) to determine the effects of modulation of the various components of the TGF-beta system on in vitro and in vivo growth properties in two independent isogenic models of colon carcinoma. Cells were stably transfected with a tetracycline-repressible RII expression vector (CBS4-RII), a tetracycline-repressible expression vector containing a truncated RII cDNA lacking the serine/threonine kinase domain (CBS4-deltaRII and FET6-deltaRII), or with a vector containing the TGF-beta1 cDNA (CBS4-beta1S and FET-beta1S). Expression of the truncated RII reduced TGF-beta sensitivity, whereas overexpression of RII increased TGF-beta sensitivity. TGF-beta overexpression did not affect TGF-beta response. In vivo tumorigenicity assays revealed that CBS4-RII cells had lower tumorigenicity than control cells, whereas CBS4-deltaRII and CBS4-beta1S had higher tumorigenicity than controls. The CBS4 cells are poorly tumorigenic in athymic mice, and the wild-type FET6 cells are nontumorigenic. FET6-deltaRII cells formed rapidly growing tumors, and FET-beta1S cells also formed tumors. These data illustrate the paradoxical tumor-promoting and -suppressing effects of TGF-beta signaling activity in two isogenic model systems from human colon carcinomas, thus demonstrating that the effects of modulation of TGF-beta expression or TGF-beta signaling capability affects malignancy in a contextual manner.

Animals↗

PubMed1999

Characterization of the effect on adhesion of different mutations in myelin P0 protein.

Table 1 summarizes the results obtained from expressing in CHO cells C21A P0 and N93A P0 alone, and each with wild-type P0. As can be seen, each of these mutated proteins reach the cell surface when expressed alone, but neither is adhesive. Finally, when each of these mutated P0 molecules is expressed with the wild-type P0, wild-type P0 is no longer adhesive. Therefore, both C21A Po and N93A P0 each have a dominant negative effect on adhesion of wild-type P0. This approach to address the functional consequences of mutations in P0 can now be used to assess the effects of different mutations associated with CMT1B.

Amino Acid Substitution↗

PubMed1999

Molecular cloning and sequencing of two 'short chain' and two 'long chain' K(+) channel-blocking peptides from the Chinese scorpion Buthus martensii Karsch.

Five full-length cDNAs encoding the precursors of two 'short chain' scorpion non-toxic peptides active on Ca(2+)-activated K(+) channels (BmP02 and BmP03) and two novel putative long chain K(+) channel-blocking peptides (named BmTXKbeta and BmTXKbeta2) were first isolated from the venom gland cDNA library of the Chinese scorpion Buthus martensii Karsch (BmK). BmTXKbeta2 showed a high similarity with AaTXKbeta, while BmTXKbeta was completely different in the deduced primary structure from the long chain and short chain scorpion toxins already characterized. Thus, BmTXKbeta expands the scorpion long chain K(+) channel-blocking peptide family. Although little sequence similarity exists between the above two short and two long peptides, they are similar at the positions of six cysteines, suggesting that they should all share a similar scaffold composed of an alpha-helix and a three-stranded beta-sheet.

3' Untranslated Regions↗

PubMed1999

Protein kinase C delta associates with and phosphorylates Stat3 in an interleukin-6-dependent manner.

Stat3 is activated by phosphorylation on Tyr-705, which leads to dimer formation, nuclear translocation, and regulation of gene expression. Serine phosphorylation of Stat3 by mitogen-activated protein kinase has also been observed in cells responding to epidermal growth factor and shown to affect its tyrosine phosphorylation and transcriptional activity. Serine phosphorylation of Stat3 is also induced by interleukin-6 (IL-6) stimulation, which is shown to be independent of mitogen-activated protein kinase and sensitive to the Ser/Thr kinase inhibitor H7. In this study, we investigated whether protein kinase C (PKC) is the kinase that is induced and responsible for Stat3 serine phosphorylation by IL-6 stimulation and which isoform of PKCs is likely to be involved. Here, we report that Stat3 was specifically associated with PKC delta in vivo in an IL-6-dependent manner in several cell types. Furthermore, Stat3 was phosphorylated by PKC delta in vivo on Ser-727, which could be inhibited either by a specific PKC delta inhibitor or by a dominant-negative mutant of PKC delta. Finally, we showed that the phosphorylation of Stat3 by PKC delta led to a negative regulation of Stat3 DNA binding and transcriptional activity. These results indicate that PKC delta is likely to be the kinase that phosphorylates Stat3 in response to IL-6 stimulation and suggest a possible regulatory role of PKC delta on Stat3 function.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1999

P-selectin expression by endothelial cells is decreased in neonatal rats and human premature infants.

Decreased adhesion of neutrophils to endothelial cells and delayed transendothelial cell migration of neutrophils have been consistently reported in neonatal animals and humans and contribute to their susceptibility to infection. The delayed transmigration of neutrophils is especially prevalent in premature neonates. To define the nature of this defect, we used an in vivo animal model of inflammation and found that radiolabeled leukocytes from adult rats transmigrated into the peritoneum of other adult rats 5 times more efficiently than they did in neonatal rats (P =.05). This indicated that defects in neonatal neutrophils could not completely account for the delayed transmigration. Delayed transmigration in the neonatal rats correlated with a defect in the expression of P-selectin on the surface of their endothelial cells. We found a similar P-selectin deficiency in endothelial cells lining mesenteric venules and umbilical veins of human premature infants when compared with term human infants. The decreased P-selectin in premature infants was associated with decreased numbers of P-selectin storage granules and decreased P-selectin transcription. Decreased P-selectin expression on the surface of endothelial cells in preterm infants may contribute to delayed neutrophil transmigration and increased susceptibility to infection.

Animals↗

PubMed1999

Statistical properties of open reading frames in complete genome sequences.

Some statistical properties of open reading frames in all currently available complete genome sequences are analyzed (seventeen prokatyotic genomes, and 16 chromosome sequences from the yeast genome). The size distribution of open reading frames is characterized by various techniques, such as quantile tables, QQ-plots, rank-size plots (Zipf's plots), and spatial densities. The issue of the influence of CG% on the size distribution is addressed. When yeast chromosomes are compared with archaeal and eubacterial genomes, they tend to have more long open reading frames. There is little or no evidence to reject the null hypothesis that open reading frames on six different reading frames and two strands distribute similarly. A topic of current interest, the base composition asymmetry in open reading frames between the two strands, is studied using regression analysis. The base composition asymmetry at three codon positions is analyzed separately. It was shown in these genome sequences that the first codon position is G- and A-rich (i.e. purine-rich); there is a co-existence of A- and T-rich branches at the second codon position; and the third codon position is weakly T-rich.

Base Sequence↗

PubMed1999

Interferon regulatory transcription factors are constitutively expressed and spatially regulated in the mouse lens.

Interferon regulatory factors (IRFs) are a family of transcription factors involved in regulation of cell growth and immunological responses. Nine IRFs have been described and they are expressed in a variety of cells, except for ICSBP and LSIRF/Pip, which are thought to be expressed exclusively in immune cells. Here, we show that IRF-1, IRF-2, ICSBP, and LSIRF/Pip are constitutively expressed in the mouse lens. These IRFs are present in both the cytoplasm and the nuclei of lens cells. However, the nuclear and cytoplasmic proteins exhibit distinct mobilities on SDS/PAGE. We further show that in the developing mouse lens, IRF-1 and IRF-2 are expressed at high levels in differentiated lens fiber cells with very low and barely detectable levels in undifferentiated lens epithelial cells. Although the level of ICSBP expression is very low in the normal mouse lens, in transgenic mice with constitutive expression of interferon gamma in the lens, its level is markedly elevated and ICSBP expression is detected exclusively in the nuclei of undifferentiated lens cells. Taken together, our data suggest that expression of IRF transcription factors is spatially regulated in the lens and that distinct IRFs may contribute to differential gene regulation in the epithelial and fiber compartments of the vertebrate lens.

Animals↗

PubMed1999

Magnesium modulates contractile responses of rat aorta to thiocyanate: A possible relationship to smoking-induced atherosclerosis.

Thiocyanate anions (SCN-) as the end products of tobacco smoke and found in the blood of cigarette smokers have been implicated in atherogenesis and heart diseases. Magnesium deficiency has also been implicated in the etiology of atherogenesis. The contractile responses of rat aorta to SCN- and the modulation of extracellular magnesium ions ([Mg2+]o) on the effect of SCN- were, therefore, studied in isolated rat aortic rings. SCN- exposure at a range of concentrations (from 10(-5) to 5 x 10(-2) M) induces contractile responses of isolated rat aortic rings with and without endothelium in a concentration-dependent manner. Significant differences in responsiveness to SCN- were found in rat aortic ring segments with and without endothelial cells. Preincubation of these vessels with low [Mg2+]o markedly shifted the contractile concentration-effect curves to the left, and the contractile effects of SCN- in rat aortic rings were potentiated. In contrast to lowering [Mg2+]o, increasing [Mg2+]o to 2.4 mM was found to dramatically attenuate the contractile responses to SCN-. In the absence of extracellular Ca2+ ([Ca2+]o), SCN--induced contractions were, however, almost abolished after exposure to Mg2+-free medium. In order to investigate the mechanisms of [Mg2+]o modulation of SCN--induced contractile response of rat aorta, changes in intracellular Ca2+ ([Ca2+]i) were measured in cultured primary smooth muscle cells isolated from rat aorta. The resting level of [Ca2+]i in the rat aortic smooth muscle cells was 80.6 +/- 6.6 nM. Exposure of these cells to SCN- (5 x 10(-5) to 5 x 10(-3) M) produced rises in [Ca2+]i in a concentration-dependent manner. Preincubation of these cells with low [Mg2+]o (0 or 0.3 mM, the lowest physiological range) for 24 h significantly potentiated increments in [Ca2+]i induced by SCN-. These rises in [Ca2+]i induced by SCN- were completely inhibited by pretreating the cells with 2.4 mM [Mg2+]o for 24 h. These results support a hypothesis whereby cigarette smoking or exposure to smoking can induce cardiovascular diseases, at least partly, probably by causing spasm and thickening of arterial blood vessels as a consequence of large rises in [Ca2+]i in vascular smooth muscle cells. The chronic presence of or exposure to both thiocyanate and low Mg2+ in the blood of smokers can result in rapid flux of Ca2+ into vascular smooth muscle cells, thus accelerating or initiating atherosclerotic processes in smokers.

Animals↗

PubMed1999

Mediation of N-(4-hydoxyphenyl)retinamide-induced apoptosis in human cancer cells by different mechanisms.

The induction of apoptosis by the synthetic retinoid N-(4-hydroxyphenyl)retinamide (4HPR) has been documented in vitro in various cancer types. A role for reactive oxygen species (ROS) in apoptosis induced by 4HPR in some cancer cells has been demonstrated recently. We studied five different human head and neck and five lung cancer cell lines to determine whether the ROS play a general role in 4HPR-induced apoptosis. We found that 4HPR induced apoptosis in all of the cell lines; however, this effect was blocked by antioxidants in only 2 of the 10 cell lines. 4HPR induced a greater than 4-fold increase in the generation of intracellular ROS in these two cell lines compared with a much lower effect in other cell lines. Furthermore, these two cell lines were most sensitive to the induction of apoptosis by 4HPR. The level of the cellular antioxidant thiol and superoxide dismutase activity were relatively lower in cells, which responded to 4HPR with a high level of ROS generation. These results indicate that although ROS can mediate 4HPR-induced apoptosis in some cells, which may have a low endogenous cellular antioxidant levels, other mechanisms exist for 4HPR-induced apoptosis. One such mechanism may involve retinoic acid receptors (RARs) because an RAR antagonist was able to block partially 4HPR-induced apoptosis. In conclusion, 4HPR-induced apoptosis involves at least three different mechanisms, which are complex and can overlap in the same cell line: (a) one mechanism involving 4HPR-induced ROS; (b) one involving RARs; and (c) at least one that does not involve ROS or RARs and remains unclear.

Antineoplastic Agents↗

PubMed1999

Pyrrolidine dithiocarbamate prevents ethanol-induced elevation of [Ca2+]i in cultured canine cerebral vascular smooth muscle cells.

Pyrrolidine dithiocarbamate (PDTC) has been shown to block nuclear transcription factor (NF-kappaB) activation induced by a wide range of stimuli in different cell lines. NF-kappaB is a common element of the promoter region of inflammatory cytokines which can be stimulated by ethanol. Recently, we have shown that PDTC can ameliorate cerebrovascular damage, brain cortical damage, leukocyte adhesion and rolling, and stroke induced by ethanol. We, therefore, tested the effects of preincubation with PDTC on alcohol-induced changes in intracellular free calcium ions ([Ca2+]i) in cultured canine cerebral smooth muscle cells. These vascular cells, chronically treated with ethanol (10-100 mM) for 24 and 72 h, exhibited concentration-dependent rises in [Ca2+]i. PDTC (0.1 microM) itself failed to influence resting levels of [Ca2+]i in these vascular muscle cells. PDTC (0.1 microM) pretreatment, however, inhibited completely the elevations in [Ca2+]i induced by chronic ethanol (10-100 mM). The present results suggest that ethanol-induced production of reactive oxygen species and elevation of [Ca2+]i in cultured canine cerebral vascular smooth muscle cells triggers induction of transcription factor NF-kappaB, which could play an important role in alcohol-induced brain damage and stroke.

Animals↗

PubMed1999

Declining HIV/AIDS mortality in New York City.

BACKGROUND: New York City has only 3% of the U.S. population but has reported nearly 16% of all AIDS cases. METHODS: This is an observational study using the New York City vital events and AIDS case surveillance registries to describe trends in HIV/AIDS mortality from 1983 through June 30, 1998. RESULTS: Annual HIV/AIDS deaths increased steadily until stabilizing at 7046 in 1995, declined 29% to 4998 in 1996, and declined 47% to 2625 in 1997. Comparing data from 1997 with those from 1995, declines occurred in all demographic groups and in all major HIV transmission categories: 74% in men who have sex with men, 68% in injecting drug users, and 64% in heterosexuals. In the first 6 months of 1998, declines were smaller than they had been in previous 6-month intervals in all demographic groups except Hispanic males and those between 35 and 44 years of age. From 1995 to the first 6 months of 1998, the number of people living with AIDS in New York City increased 22% (from 32,692 to 39,976). CONCLUSIONS: The precipitous 63% decline in HIV/AIDS deaths from 1995 to 1997 occurred at the same time that more effective antiretroviral therapies became widely available. The slowing in the mortality decline observed in 1998, however, suggests that although these new therapies may have a profound effect at the population level, deaths due to AIDS will continue.

Acquired Immunodeficiency Syndrome↗

PubMed1999

Signaling pathways activated by leukocyte function-associated Ag-1-dependent costimulation.

LFA-1 binding to ICAM-1 can enhance TCR-dependent proliferation of T cells, but it has been difficult to distinguish contributions from increased adhesion, and thus TCR occupancy, versus costimulatory signaling. Whether LFA-1 ligation results in generation of a unique costimulatory signal(s) distinct from those activated by the TCR has been unclear. Using purified ligands, it is shown that ICAM-1 and B7. 1 provide comparable costimulation for proliferation of CD8+ T cells, and that both ligands up-regulate the activities of phosphatidylinositol 3-kinase, sphingomyelinase, and c-Jun NH2-terminal kinase (JNK). These pathways are distinct from those activated by the TCR, and have previously been implicated in up-regulating IL-2 production in response to CD28-B7 interaction. Thus, under conditions in which ICAM-1 provides costimulation of proliferation, LFA-1 ligation activates some of the same signaling pathways as does CD28 ligation. LFA-1 and CD28 do not act identically, however, as indicated by differential sensitivity to inhibitors of phosphatidylinositol 3-kinase; LFA-1-dependent costimulation of proliferation is inhibited, while CD28-dependent costimulation is not. Given the broad distribution of class I and ICAMs on many cell types, the ability of LFA-1 to provide costimulatory signals has implications for where and how CD8+CTL may become activated in response to an antigenic challenge.

Androstadienes↗

PubMed1999

[Polymorphism of HLA-DRB1, DQB1 in the Hans of north China]

OBJECTIVE: To understand polymorphism of HLA-DRB1, DQB1 in the Hans of north China and obtain more comprehensive and accurate data on genetics at DNA level. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) were used to determine HLA-DRB1, DQB1 alleles in 107 unrelated healthy Han individuals of north China. RESULTS: The authors determined 14 DRB1, 9 DQB1 alleles, which included not only the allele frequencies that corresponded to the gene frequencies of DR, DQ loci determined by other 9 cooperating labs but also the allele frequencies of DRB1*15, DRB1*16, DQB1*0301, DQB1*0302, DQB1*0303, DQB1*05, DQB1*0601, DQB1*0602, and DQB1*0604 that other serology labs did not report. CONCLUSION: This study has obtained a more comprehensive and accurate data set of the normal allele frequencies and linkage disequilibrium parameters of HLA-DRB1,DQB1 in the Hans of north China, which may be of significance in the studies on population genetics and disease association.

Journal Article↗

PubMed1999

The expression of GABA(A) receptor alpha2 subunit is upregulated by testosterone in rat cerebral cortex.

The GABAergic system is sexually dimorphic in certain brain regions and can be regulated by testosterone (T). However, the contribution of T to sex-specific developmental processes in the brain is less clear. We have examined whether T regulates expression of GABA(A) receptor alpha2 subunit in the cerebral cortex of embryonic and postnatal female rats using in situ hybridization and Western blotting. We found that both alpha2 mRNA and protein levels are significantly increased by T treatment at embryonic day 20 (E20) and birth (P0). The observed modulation of the expression of GABA(A) receptor alpha2 subunit by T may be translated into changes in the levels or composition of GABA(A) receptor, either of which would be expected to alter neuronal functional response to GABA activation. As the effects of T are developmental-stage-specific, they may have an organizational impact on brain development.

Analysis of Variance↗

PubMed1999

Interferon consensus sequence-binding protein is constitutively expressed and differentially regulated in the ocular lens.

Interferon signaling is mediated by STATs and interferon regulatory factor (IRF) families of transcription factors. Ten distinct IRFs have been described and most are expressed in a variety of cells except for interferon consensus sequence-binding protein (ICSBP) and lymphoid-specific IRF/Pip that are thought to be exclusively expressed in lymphoid cells. We show here for the first time that ICSBP is constitutively and inducibly expressed in the mouse lens. In contrast to lymphoid cells with exclusive expression of ICSBP in the nucleus, ICSBP is present in both the cytoplasm and nucleus of the lens cell. However, ICSBP in the nucleus is of lower apparent molecular weight. We further show that the ICSBP promoter is constitutively bound by lens nuclear factors and that its activation requires binding of additional factors including STAT1. Furthermore, transcriptional activation of ICSBP gene by interferon gamma is accompanied by selective nuclear localization of ICSBP in proliferating epithelial cells but not in the nuclei of nondividing cells in the lens fiber compartment. Constitutive and inducible expression of ICSBP in the ocular lens and differential regulation of its subcellular localization in the developing lens suggest that ICSBP may have nonimmunity related functions and that the commonly held view that it is lymphoid-specific be modified.

Animals↗

PubMed1999

Heparin-induced conformational change in microtubule-associated protein Tau as detected by chemical cross-linking and phosphopeptide mapping.

In Alzheimer's disease, microtubule-associated protein tau becomes abnormally phosphorylated and aggregates into paired helical filaments. Sulfated glycosaminoglycans such as heparin and heparan sulfate were shown to accumulate in pretangle neurons, stimulate in vitro tau phosphorylation, and cause tau aggregation into paired helical filament-like filaments. The sulfated glycosaminoglycan-tau interaction was suggested to be the central event in the development of neuropathology in Alzheimer's disease brain (Goedert, M., Jakes, R., Spillantini, M. G., Hasegawa, M., Smith, M. J., and Crowther, R. A. (1996) Nature 383, 550-553). The biochemical mechanism by which sulfated glycosaminoglycans stimulate tau phosphorylation and cause tau aggregation remains unclear. In this study, disuccinimidyl suberate (DSS), a bifunctional chemical cross-linker, cross-linked tau dimers, tetramers, high molecular size aggregates, and two tau species of sizes 72 and 83 kDa in the presence of heparin. In the absence of heparin only dimeric tau was cross-linked by DSS. Fast protein liquid chromatography gel filtration revealed that 72- and 83-kDa species were formed by intramolecular cross-linking of tau by DSS. These observations indicate that heparin, in addition to causing aggregation, also induces a conformational change in tau in which reactive groups are unmasked or move closer leading to the DSS cross-linking of 72- and 83-kDa species. Heparin-induced structural changes in tau molecule depended on time of heparin exposure. Dimerization and tetramerization peaked at 48 h, whereas conformational change was completed within 30 min of heparin exposure. Heparin exposure beyond 48 h caused an abrupt aggregation of tau into high molecular size species. Heparin stimulated tau phosphorylation by neuronal cdc2-like kinase (NCLK) and cAMP-dependent protein kinase. Phosphopeptide mapping and phosphopeptide sequencing revealed that tau is phosphorylated by NCLK on Thr212 and Thr231 and by cAMP-dependent protein kinase on Ser262 only in the presence of heparin. Heparin stimulation of tau phosphorylation by NCLK showed dependence on time of heparin exposure and correlated with the heparin-induced conformational change of tau. Our data suggest that heparin-induced conformational change exposes new sites for phosphorylation within tau molecule.

Amino Acid Sequence↗

PubMed1999

VEGF prevents apoptosis of human microvascular endothelial cells via opposing effects on MAPK/ERK and SAPK/JNK signaling.

Vascular endothelial growth factor (VEGF), an endothelial cell-specific mitogen, promotes endothelial cell survival and angiogenesis. We recently showed that VEGF can support the growth of human dermal microvascular endothelial cells (HDMEC) and human umbilical vein endothelial cells in serum-free medium. Reasoning that VEGF might be modulating apoptotic signal transduction pathways, we examined mechanisms involved in the anti-apoptotic effect of VEGF on starvation- and ceramide-induced apoptosis in HDMEC. We observed that VEGF ameliorated the time-dependent increase in apoptosis, as demonstrated by morphologic observations, TUNEL assay, and DNA fragmentation. On the other hand, basic fibroblast growth factor only partially prevented apoptosis in serum-starved HDMEC; platelet-derived growth factor-BB was completely ineffective. VEGF activated the phosphorylation of extracellular signal regulated kinase (ERK)1 (p44 mitogen-activated protein kinase; MAPK) and ERK2 (p42 MAPK) in a time- and concentration-dependent manner. Both the VEGF-induced activation and its anti-apoptotic effect were prevented by the specific MAPK/ERK inhibitor PD98059. The presence of VEGF also inhibited the sustained activation of stress-activated protein kinase/c-jun-NH2-kinase (SAPK/JNK) caused by serum starvation and ceramide treatment. Activation of the MAPK pathway together with inhibition of SAPK/JNK activity by VEGF appears to be a key event in determining whether an endothelial cell survives or undergoes programmed cell death.

Apoptosis↗