PMID- 27471855
OWN - NLM
STAT- MEDLINE
DCOM- 20170802
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 7
DP  - 2016
TI  - Granular Formation during Apoptosis in Blastocystis sp. Exposed to Metronidazole 
      (MTZ).
PG  - e0155390
LID - 10.1371/journal.pone.0155390 [doi]
LID - e0155390
AB  - The role and function of the granular life cycle stage in Blastocystis sp, 
      remains uncertain despite suggestions being made that the granules are metabolic, 
      reproductive and lipid in nature. This present study aims to understand granular 
      formation by triggering apoptosis in Blastocystis sp. by treating them with 
      metronidazole (MTZ). Blastocystis sp.cultures of 4 sub-types namely 1, 2, 3 and 5 
      when treated with 0.01 and 0.0001 mg/ml of metronidazole (MTZ) respectively 
      showed many of the parasites to be both viable and apoptotic (VA). Treated 
      subtype 3 isolates exhibited the highest number of granular forms i.e. 88% 
      (p<0.001) (0.0001 mg/ml) and 69% (p<0.01) (0.01 mg/ml) respectively at the 72 h 
      in in vitro culture compared to other subtypes. These VA forms showed distinct 
      granules using acridine orange (AO) and 4',6-diamino-2-phenylindole (DAPI) 
      staining with a mean per cell ranging from 5 in ST 5 to as high as 16 in ST 3. 
      These forms showed intact mitochondria in both viable apoptotic (VA) and viable 
      non-apoptotic (VNA) cells with a pattern of accumulation of lipid droplets 
      corresponding to viable cells. Granular VA forms looked ultra-structurally 
      different with prominent presence of mitochondria-like organelle (MLO) and a 
      changed mitochondrial trans-membrane potential with thicker membrane and a highly 
      convoluted inner membrane than the less dense non-viable apoptotic (NVA) cells. 
      This suggests that granular formation during apoptosis is a self-regulatory 
      mechanism to produce higher number of viable cells in response to treatment. This 
      study directs the need to search novel chemotherapeutic approaches by 
      incorporating these findings when developing drugs against the emerging 
      Blastocystis sp. infections.
FAU - Dhurga, Devi Balkrishnan
AU  - Dhurga DB
AUID- ORCID: 0000-0001-9968-1843
AD  - Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, 
      Kuala Lumpur, Malaysia.
FAU - Suresh, Kumar
AU  - Suresh K
AD  - Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, 
      Kuala Lumpur, Malaysia.
FAU - Tan, Tian Chye
AU  - Tan TC
AD  - Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, 
      Kuala Lumpur, Malaysia.
LA  - eng
PT  - Journal Article
DEP - 20160729
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (4',6-diamino-2-phenylindole)
RN  - 0 (Antiprotozoal Agents)
RN  - 0 (Diamines)
RN  - 0 (Indoles)
RN  - 140QMO216E (Metronidazole)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/chemistry
MH  - Animals
MH  - Antiprotozoal Agents/*pharmacology
MH  - Apoptosis/*drug effects
MH  - Blastocystis/*drug effects/metabolism
MH  - Diamines/chemistry
MH  - Indoles/chemistry
MH  - Lipid Metabolism
MH  - Membrane Potential, Mitochondrial/drug effects
MH  - Metronidazole/*pharmacology
MH  - Microscopy, Electron, Transmission
PMC - PMC4966910
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/07/30 06:00
MHDA- 2017/08/03 06:00
PMCR- 2016/07/29
CRDT- 2016/07/30 06:00
PHST- 2016/01/19 00:00 [received]
PHST- 2016/04/04 00:00 [accepted]
PHST- 2016/07/30 06:00 [entrez]
PHST- 2016/07/30 06:00 [pubmed]
PHST- 2017/08/03 06:00 [medline]
PHST- 2016/07/29 00:00 [pmc-release]
AID - PONE-D-16-00564 [pii]
AID - 10.1371/journal.pone.0155390 [doi]
PST - epublish
SO  - PLoS One. 2016 Jul 29;11(7):e0155390. doi: 10.1371/journal.pone.0155390. 
      eCollection 2016.