Apoptosis and Autophagy Function Cooperatively for the Efficacious Execution of Programmed Nurse Cell Death During Drosophila virilis Oogenesis

Programmed cell death consists of two major types, apoptotic and autophagic, both of which are mainly defined by morphological criteria. Our findings indicate that both types of programmed cell death occur in the ovarian nurse cells during middle and late oogenesis of Drosophila virilis. During mid-oogenesis, the spontaneously degenerated egg chambers exhibit typical characteristics of apoptotic cell death. Their nurse cells contain condensed chromatin and fragmented DNA, whereas active caspase assays and immunostaining procedures demonstrate the presence of highly activated caspases. Distinct features of autophagic cell death are also observed during D. virilis mid-oogenesis, as shown by monodansylcadaverine staining and ultrastructural examination performed by transmission electron microscopy. Additionally, atretic egg chambers exhibit an accumulation of lysosomal proteases. At the late stages of D. virilis oogenesis, apoptosis and autophagy coexist, manifesting cell death features that are similar to the ones described above, being also escorted by the involvement of an altered cytochrome c conformational display. We propose that apoptosis and autophagy operate synergistically during D. virilis oogenesis for a more efficient elimination of the degenerated nurse cells.

Addendum to:

Mechanisms of Programmed Cell Death During Oogenesis in Drosophila virilis

A.D. Velentzas, I.P. Nezis, D.J. Stravopodis, I.S. Papassideri and L.H. Margaritis

Cell Tissue Res 2006; doi: 10.1007/s00441-006-0298-x     

Early Miocene rodents of Gökler (Kazan Basin,Central Anatolia,Turkey)

Abstract

The rich and relatively diverse fossil mammalian assemblage from Gökler is of special importance for understanding of faunal evolution in Central Anatolia. Large mammals were not recovered, but insectivores and rodents are abundant. The assemblage of rodents is studied in detail and comprises mainly diversified cricetids. Dormice are abundant, but are represented by only one species. Squirrels are represented only by few specimens and also beaver remains were identified. Spanocricetodon sinuosus is referred to a new genus Latocricetodon nov. gen that is tentatively assigned to the Pseudocricetodontinae. Newly named species are Cricetodon goklerensis sp. nov., Democricetodon haltmari sp. nov., Eumyarion lukasi sp. nov. and Glirudinus matusi sp. nov. The rodent assemblage is assigned to local zone C which is correlated to the European biounit MN2 (early Miocene). Our biochronological assessment is supported by radiometric dating from two volcanic ash layers.

Latocricetodon

LSID http://zoobank.org/urn:lsid:zoobank.org:act:3414DB1E-0C5E-4154-BE5E-02A9ED183B1A

Cricetodon goklerensis

LSID http://zoobank.org/urn:lsid:zoobank.org:act:1B658872-6C10-4355-B87C-3E6277AF4EDA

Democricetodon haltmari

LSID http://zoobank.org/urn:lsid:zoobank.org:act:9B13F956-7F8C-406A-9970-1F5E999E54C6

Eumyarion lukasi

LSID http://zoobank.org/urn:lsid:zoobank.org:act:34DED87E-855F-4969-AB84-10BED5C572BF

Glirudinus matusi

LSID http://zoobank.org/urn:lsid:zoobank.org:act:798ECB9A-E3B5-4C38-B5B3-FEB17AF734FE     

An interview with Dr. John J. Tyson on his highly cited paper published in Cell Cycle

Comment on: Steady states and oscillations in the p53/Mdm2 network.

Ciliberto A, et al. Cell Cycle 2005; 4:488-93.     

CDK2-Activating Kinase (CAK): More Questions than Answers

Commentary to:

The cyclin-dependent kinase (CDK) Family member PNQALRE/CCRK supports cell proliferation but has no intrinsic CDK-activating kinase (CAK) activity.

Lara Wohlbold, Stéphane Larochelle, Jack C.-F. Liao, Geulah Livshits, Juliet Singer, Kevan M. Shokat and Robert P. Fisher

Cell Cycle. 2006 Mar;5(5):546-54.     

Autophagy During Conidiation and Conidial Germination in Filamentous Fungi

Filamentous fungi form aerial hyphae on solid medium, and some of these differentiate into conidiophores for asexual sporulation (conidiation). In the filamentous deuteromycete, Aspergillus oryzae, aerial hyphae are formed from the foot cells and some differentiate into conidiophores, which are composed of vesicles, phialides and conidia. Recently, we isolated the yeast ATG8 gene homologue Aoatg8 from A. oryzae, and visualized autophagy by the expression of an EGFP (enhanced green fluorescent protein)–AoAtg8 fusion protein and DsRed2 protein in this fungus. Furthermore, by constructing the Aoatg8 deletion and conditional mutants, we demonstrated that autophagy functions during the process of differentiation of aerial hyphae, conidiation and conidial germination in A. oryzae. Here, we discuss the contribution of autophagy towards the differentiation and germination processes in filamentous fungi.

Addendum to:

Functional Analysis of the ATG8 Homologue Aoatg8 and Role of Autophagy in Differentiation and Germination in Aspergillus oryzae

T. Kikuma, M. Ohneda, M. Arioka and K. Kitamoto

Eukaryot Cell 2006; 5:1328-36     

Calpain as a Novel Regulator of Autophagosome Formation

Ubiquitously expressed mu- and m-calpain proteases consist of 80-kDa catalytic subunits encoded by the Capn1 and Capn2 genes, respectively, and a common 28-kDa regulatory subunit encoded by the calpain small 1 (Capns1) gene.

The mu- and m-calpain proteases have been implicated in both pro- or anti-apoptotic functions. We have found that Capns1 depletion is coupled to increased sensitivity to apoptosis triggered by a number of autophagy-inducing stimuli in mammalian cells. Therefore we investigated the involvement of calpains in autophagy using MEFs derived from Capns1 knockout mice and Capns1 depleted human cells as model systems.

We found that autophagy is impaired in Capns1-deficient cells by immunostaining of the endogenous autophagosome marker LC3 and electron microscopy experiments. Accordingly, the enhancement of lysosomal activity and long-lived proteins degradation, normally occurring upon starvation, are also reduced.

In Capns1-depleted cells ectopic LC3 accumulates in early endosome-like vesicles that might represent a salvage pathway for protein degradation when autophagy is defective.

Addendum to:

Calpain is Required for Macroautophagy in Mammalian Cells

Francesca Demarchi, Cosetta Bertoli, Tamara Copetti, Isei Tanida, Claudio Brancolini, Eeva-Liisa Eskelinen and Claudio Schneider

J Cell Biol 2006; 175:595-605     

Autophagy During Conidiation,Conidial Germination and Turgor Generation in Magnaporthe grisea

Autophagy is a ubiquitous and evolutionarily conserved process found in all eukaryotic cells that allows for the degradation and recycling of old proteins and organelles. Starvation can induce autophagy, and autophagic pathway is an essential process for cellular function under starvation. In Magnaporthe grisea, starvation is one of the key induced factors for the germ tube tip to differentiate into an appressorium. Considering the importance of the rice blast fungus as a primary model for host-pathogen interaction, the role of autophagy in fungal development, appressorium turgor generation and pathogenicity of M. grisea via its role in organelle and protein turnover is a very significant subject.

Addendum to:

Involvement of a Magnaporthe grisea Serine/Threonine Kinase, MgATG1, in Appressorium Turgor and Pathogenesis

X.-H. Liu, J.-P. Lu, L. Zhang, B. Dong, H. Min and F.-C. Lin

Eukaryotic Cell 2007; In press     

Atg9 Trafficking in Yeast Saccharomyces cerevisiae

Autophagy can be divided into selective and non-selective modes. This process is considered selective when a precise cargo is specifically and exclusively incorporated into autophagosomes, the double-membrane vesicles that are the hallmark of autophagy. In contrast, during nonselective, bulk autophagy, cytoplasmic components are randomly enwrapped into autophagosomes. To date, approximately 30 autophagy-related genes called ATG have been identified. Sixteen of them compose the general basic machinery catalyzing the formation of double-membrane vesicles in all eukaryotic cells. The rest of them are often not conserved between species and cooperate with the basic Atg proteins during either selective or nonselective autophagy. Atg9 is the only integral membrane component of the conserved Atg machinery and appears to be a crucial organizational element.5 Recent studies in the S. cerevisiae have shown that Atg9 transport is differentially regulated depending on the autophagy mode. In this addendum, we will review and discuss what has recently been unveiled about yeast S. cerevisiae Atg9 trafficking, its modulators and its potential role in double-membrane vesicle biogenesis.

Addendum to:

Atg9 Sorting from Mitochondria is Impaired in Early Secretion and VFT Complex Mutants in Saccharomyces cerevisiae

F. Reggiori and D.J. Klionsky

J Cell Sci 2006: 119:2903-11     

Some Notes on the Causes of Bird Road Casualties

Guide des Oiseaux d'Europe, by R. Peterson, G. Mountfort and P. Hollom, translated by P. Géroudet. Delachaux &; Niestlé, 1967.

A Field Guide to the Mammals of Britain and Europe, by F. H. Van den Brink. Collins, 1967. 30/-. 221 pages, 32 plates, numerous text-figs. and maps.

Hours and the Birds, by R. D. Symons. Toronto University Press. London: Oxford University Press, 1968. £5 19s. 224 pages, 17 coloured plates, text-figs.

Galapagos: Islands of Birds, by Bryan Nelson. Longmans, 1968.50/-. 338 pages, 57 photos, 56 text-figs.

A la Découverte de la Nature, by O. Paccaud. Delachaux &; Niestlé, Neuchatel, Switzerland, 1967. 442 pages, numerous illustrations.

On Predation and Life, by Paul L. Errington. Iowa State University Press, 1967. 277 pages, numerous black and white illustrations.

The Courtship Habits of the Great Crested Grebe, by Julian Huxley. Jonathan Cape, London, 1968. 7/6 (paperback), 18/- (hardback). 98 pages, text-figures.

Birds of South Vietnam, by Philip Wildash. Charles E. Tuttle Co., Inc., Rutland, Vermont, 1968 (Agents in U.K.: Prentice-Hall International, London). 70/-. 234 pages, 25 colour plates, 21 black and white drawings.

The Psychology of Birds: An interpretation of Bird Behaviour, by Harold E. Burtt. Collier-MacMillan Ltd., London, 1968. 55/-. 242 pages, 11 line drawings.     

Autophagy-Related Pathways and Specific Role of Sterol Glucoside in Yeasts

Recently, we showed that the requirement of sterol glucoside (SG) during pexophagy in yeasts is dependent on the species and the nature of peroxisome inducers. Atg26, the enzyme that converts sterol to SG, is essential for degradation of very large methanol-induced peroxisomes, but only partly required for degradation of smaller-sized oleate- and amine-induced peroxisomes in Pichia pastoris. Moreover, oleate- and amine-induced peroxisomes of another yeast, Yarrowia lipolytica, are degraded by an Atg26-independent mechanism. The same is true for degradation of oleate-induced peroxisomes in Saccharomyces cerevisiae. Here, we review our findings on the specificity of Atg26 function in pexophagy and extend our observations to the role of SG in the cytoplasm to vacuole targeting (Cvt) pathway and bulk autophagy. The results presented here and elsewhere indicate that Atg26 might increase the efficacy of all autophagy-related pathways in P. pastoris, but not in other yeasts. Recently, it was shown that P. pastoris Atg26 (PpAtg26) is required for elongation of the pre-autophagosomal structure (PAS) into the micropexophagic membrane apparatus (MIPA) during micropexophagy. Therefore, we speculate that SG might facilitate elongation of any double membrane from the PAS and this enhancer function of SG becomes essential when extremely large double membranes are formed.

Addendum to:

The Requirement of Sterol Glucoside for Pexophagy in Yeast Is Dependent on the Species and Nature of Peroxisome Inducers

T.Y. Nazarko, A.S. Polupanov, R.R. Manjithaya, S. Subramani and A.A. Sibirny

Mol Biol Cell 2007; 18:106-18     

Functions of PI4P and Sterol Glucoside Necessary for the Synthesis of a Nascent Membrane Structure During Pexophagy

We recently showed that, in the yeast Pichia pastoris, an ergosterol glucoside synthesizing enzyme Atg26 is recruited to the precursor of the pexophagic structure, micropexophagic membrane apparatus (MIPA), under the regulation of phosphatidylinositol 4'-monophosphate (PI4P)-signaling during pexophagy. Atg26 was found to harbor a novel PI4P-binding motif, GRAM domain. Both lipids, PI4P and sterol glucoside, synthesized by PpPik1 and PpAtg26, respectively, were necessary for pexophagy, in the step where the MIPA was formed. In this addendum, we review these findings, and speculate the mechanistic and physiological implications of the functions of these lipids during the autophagic process.

Addendum to:

PI4P-Signaling Pathway for the Synthesis of a Nascent Membrane Structure in Selective Autophagy

S. Yamashita, M. Oku, Y. Wasada, Y. Ano and Y. Sakai

J Cell Biol 2006; 173: 709-17     

Serum tumor markers in bile duct cancer – a review

Abstract

Context: Bile duct cancer (BDC) is a disease with a very grave prognosis, often diagnosed too late.

Objective: The aim of this review is to evaluate available literature on tumor markers in serum from patients with BDC.

Methods: Using the search words “serum markers”, “bile duct cancer”, “cholangiocarcinoma”, “biomarker” and “tumor marker”, a search was carried out.

Results: Seventy-five studies were included in the review.

Conclusion: CA19-9 is by far the most studied and most promising diagnostic and/or prognostic marker in BDC. But also the different mucins are interesting as new markers of BDC in serum.     

PKCε paves the way for prostate cancer

Comment on: Benavides F, et al. Cell Cycle 2011; 10:268-77

     

Synthesis,DNA/BSA binding studies and in vitro biological assay of nickel(II) complexes incorporating tridentate aroylhydrazone and triphenylphosphine ligands

Abstract

Two new nickel (II) triphenylphosphine complexes derived from tridentate aroylhydrazone ligands [H₂L¹ = 2-hydroxy-3-methoxybenzylidene)benzohydrazone and H₂L² = N′-(2-hydroxy-3-methoxybenzylidene)-2-hydroxybenzoylhydrazone] and triphenylphosphine were prepared and their molecular structures were determined by single crystal X-ray diffraction analysis. Both nickel(II) complexes showed slightly distorted square planar geometry with one tridentate aroylhydrazone ligand coordinated through ONO donor atoms and one triphenylphosphine ligand coordinated to the nickel center through the phosphorus atom. DNA interaction studies indicated that both complexes possessed higher affinity to herring sperm DNA (HS-DNA) than the corresponding free aroylhydrazone ligand. Molecular docking investigations showed that both complexes could bind to DNA through intercalation of the phenyl rings between adjacent base pairs in the double helix. Meanwhile, bovine serum albumin (BSA) binding studies revealed the complexes could effectively interact with BSA and change the secondary structure of BSA. Further pharmacological evaluations of the synthesized complexes by in vitro antioxidant assays demonstrated high antioxidant activity against NO· and O₂˙^? radicals. The anticancer activity of each complex was assessed through in vitro cytotoxicity assays (CCK-8 kit) toward A549 and MCF-7 cancer cell and normal L-02 cell lines. Significantly, the Ni(II) complex derived from H₂L¹ ligand was found to be more effective cytotoxic toward MCF-7cancerous cell with the IC₅₀ value equaled 9.7?μM, which showed potent cytotoxic activity over standard drug cisplatin.

Abbreviations A549 human lung carcinoma cell

BSA bovine serum albumin

CCK-8 Cell Counting Kit-8

DFT density functional theory

DNA deoxyribonucleic acid

DPPH˙ 2,2-diphenyl-1-picrylhydrazyl

H₂L¹ 2-hydroxy-3-methoxybenzylidene)benzohydrazone N′-(2-hydroxy-3-methoxybenzylidene)-2-hydroxybenzoylhydrazone

H₂L² N′-(2-hydroxy-3-methoxybenzylidene)-2-hydroxybenzoylhydrazone

HOMO highest occupied molecular orbital

IC₅₀ the 50% activity

L-02 human normal liver cell

LOMO lowest unoccupied molecular orbital (LUMO)

MCF-7 human breast carcinoma cell

NO˙ nitric oxide

O₂˙^? superoxide anion

SOD superoxide dismutase

Communicated by Ramaswamy H. Sarma     

Reviews

The Birds of Glamorgan, edited by A. Heathcote, D. Griffin and H. Morrey Salmon. Cardiff Naturalists' Society, 1967 (Annex to vol. 94 of the Transactions of the Society). 30/-. 143 pages, 9 black and white photos, end-paper maps.

Birds of Norfolk, by M. J. Seago. Jarrold & Sons, Ltd., Norwich, 1967. 45 /-. 148 pages, 65 coloured photos, end-paper maps.

The Birds of Huntingdonshire, by C. F. Tebbutt. Published by the author, 1967. 75 pages, map.

A Supplement to the Birds of Cheshire, by T. Hedley Bell. Published by the author (12/16 Booth St., Manchester 2), 1967. 91 pages.

The Birds of the Doncaster District, by R. J. Rhodes. Doncaster and District Orn. Soc. (125 Chequer Avenue, Belle Vue, Doncaster), 1967. 10/-. 101 pages, map.

Pesticides and Pollution, by Kenneth Mellanby. Collins, New Naturalist, London, 1967. 30 /-. 221 pages, 30 photos, 4 in colour.

Man and Environment, by Robert Arvill. Penguin Books, London, 1967. 8/6. 332 pages, 13 black and white photos.

The Wreck of the Torrey Canyon, by Crispin Gill, Frank Booker and Tony Soper. Davis & Charles, Newton Abbott, 1967. 21/-. 128 pages, 31 black and white photos.

Courtship: a zoological study, by Margaret Bastock. Heinemann, London, 1967. 25 /-. 220 pages, 63 text-figures.

The Book of the American Woodcock, by W. G. Sheldon. University of Massachusetts Press, Amherst, Mass., 1967. $8.50. 227 pages,58 figures.

The Whooping Crane: a fight against extinction, by Faith McNulty. Longmans, Green & Co., London, 1967. 30/-. 190 pages, 26 illustrations.

Birds of Australia, A. Rutgers. 2 vols. Methuen, London, 1967. 35/- each. Each with 80 coloured plates from the lithographs of John Gould.     

CYFRA21-1 can predict the sensitivity to chemoradiotherapy of non-small-cell lung carcinoma

Background: The increasing panel of systemic therapies enables the individual management of lung cancer patients, even in advanced stages. However, predictive tools indicating the efficacy of chemoradiotherapy (CRT) are badly needed.

Aims: To determine the tumour markers for predicting the therapeutic effect in non-small-cell lung carcinoma (NSCLC) patients treated with CRT.

Methods: The serum levels of cytokeratin 19 fragment antigen 21-1 (CYFRA21-1), neurone-specific enolase (NSE) and carcinoembryonic antigen (CEA) were measured before CRT by enzyme-linked immunosorbent assays, while the tumour responses were assessed according to the World Health Organization (WHO) response criteria. The relationships between pretreatment expression of CYFRA21-1, NSE, CEA and the effectiveness of CRT were analysed.

Results: The complete response (CR) rate of the primary tumours estimated by computed tomography in patients with high levels of CYFRA21-1 was 2.9% (2/68) while in cases with low CYFRA21-1 it was 20.3% (12/59) (p?=?0.005). The effective rates (CR+PR) in CYFRA21-1 high and low groups were 52.9% (36/68) and 72.9% (43/59), respectively (p?=?0.022).

Conclusions: CYFRA21-1 may be a reliable surrogate marker of CRT efficacy in patients with NSCLC.     

Maintaining T Lymphocyte Homeostasis: Another Duty of Autophagy

First identified as a pathway for nutrient recovery during periods of starvation, the role of autophagy has expanded to the clearance of “toxic” intracellular material including ubiquitin-positive protein aggregates, damaged organelles as well as microbial pathogens in various cell types. We have examined the role of autophagy in the development and function of the adaptive immune system. Genes encoding autophagy machinery are expressed in T lymphocytes, and autophagy occurs in primary CD4⁺ and CD8⁺ T cells. By generating fetal liver chimeric mice, we found that thymocyte development is largely normal but the mature T cell compartment is severely reduced in the absence of the essential autophagy gene Atg5. Consistent with a critical role for autophagy in promoting T cell survival, Atg5^-/- CD8⁺ T cells display high levels of apoptosis. Surprisingly, Atg5-deficient T cells were also unable to efficiently proliferate after T-cell receptor (TCR) stimulation. These findings suggest that autophagy regulates T lymphocyte homeostasis by promoting both survival and proliferation. In addition, T cells offer a new, physiologically relevant system to study the regulation and function of autophagy pathways in vivo.

Addendum to:

A Critical Role for the Autophagy Gene Atg5 in T Cell Survival and Proliferation

H.H. Pua, I. Dzhagalov, M. Chuck, N. Mizushima and Y.W. He

J Exp Med 2007; 204:25-31     

C-Phycocyanin elicited antitumor efficacy via cell-cycle arrest,apoptosis induction,and invasion inhibition in esophageal squamous cell carcinoma

Objectives: Mounting evidence has demonstrated that C-Phycocyanin (C-PC) exhibits marked antitumor activity in a wide type of tumors, such as pancreas cancer, breast carcinoma, lung cancer, and colon cancer. The current study aimed to confirm the antitumor efficacy of C-PC in esophageal squamous cell carcinoma (ESCC).

Methods: The efficacy of C-PC was evaluated against the proliferation of ESCC cell lines EC9706 and EC1 by CCK-8 kit and in a mice model of ESCC EC9706. Cell cycle and apoptosis were investigated by flow cytometry, and cell invasion was determined via transwell chamber. Protein expression was examined by Western blots.

Results: We found that C-PC exhibited anti-proliferation ability in a time-dependent manner and a dose-dependent manner in ESCC EC9706 and EC1 cells. Besides, C-PC markedly arrested cell cycle in the G0/G1 phase, induced cell apoptosis and suppressed cell invasion ability in both EC9706 and EC1 cells (p?<?.01). Notably, C-PC evoked the elevations of Bax, PARP, and cleaved-caspase-3 protein, but reduced cyclin D1, CDK4, Bcl-2, MMP-2, and MMP-9 expression levels. Further investigation from in vivo experiment revealed that C-PC displayed significant antitumor efficacy in the xenografted EC9706 model.

Conclusions: Our data presented herein suggest C-PC exerts antitumor efficacy in ESCC.     

NFκB anti-apoptotic or pro-apoptotic,maybe both

Comment on: Martin AG, et al. Aging 2009;1:335-49.

And Jim O’Prey, et al. Cell Cycle 2010; 9:947-52.