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1.  Comparison between open and closed methods of herniorrhaphy in calves affected with umbilical hernia 
Journal of Veterinary Science  2009;10(4):343-347.
Umbilical hernias in calves commonly present to veterinary clinics, which are normally secondary to failure of the normal closure of the umbilical ring, and which result in the protrusion of abdominal contents into the overlying subcutis. The aim of this study was to compare the suitability of commonly-used herniorrhaphies for the treatment of reducible umbilical hernia in calves. Thirty-four clinical cases presenting to the Veterinary Teaching Hospital, Chittagong Veterinary and Animal Sciences University, Chittagong, Bangladesh from July 2004 to July 2007 were subjected to comprehensive study including history, classification of hernias, size of the hernial rings, presence of adhesion with the hernial sacs, postoperative care and follow-up. They were reducible, non-painful and had no evidence of infection present on palpation. The results revealed a gender influence, with the incidence of umbilical hernia being higher in female calves than in males. Out of the 34 clinical cases, 14 were treated by open method of herniorrhaphy and 20 were treated by closed method. Complications of hernia were higher (21%) in open method-treated cases than in closed method-treated cases (5%). Hernia recurred in three calves treated with open herniorrhaphy within 2 weeks of the procedure, with swelling in situ and muscular weakness at the site of operation. Shorter operation time and excellent healing rate (80%) were found in calves treated with closed herniorrhaphy. These findings suggest that the closed herniorrhaphy is better than the commonly-used open method for the correction of reducible umbilical hernia in calves.
doi:10.4142/jvs.2009.10.4.343
PMCID: PMC2807272  PMID: 19934601
calves; herniorrhaphy; reducible umbilical hernia
2.  Strategies to Avoid Wrongly Labelled Genomes Using as Example the Detected Wrong Taxonomic Affiliation for Aeromonas Genomes in the GenBank Database 
PLoS ONE  2015;10(1):e0115813.
Around 27,000 prokaryote genomes are presently deposited in the Genome database of GenBank at the National Center for Biotechnology Information (NCBI) and this number is exponentially growing. However, it is not known how many of these genomes correspond correctly to their designated taxon. The taxonomic affiliation of 44 Aeromonas genomes (only five of these are type strains) deposited at the NCBI was determined by a multilocus phylogenetic analysis (MLPA) and by pairwise average nucleotide identity (ANI). Discordant results in relation to taxa assignation were found for 14 (35.9%) of the 39 non-type strain genomes on the basis of both the MLPA and ANI results. Data presented in this study also demonstrated that if the genome of the type strain is not available, a genome of the same species correctly identified can be used as a reference for ANI calculations. Of the three ANI calculating tools compared (ANI calculator, EzGenome and JSpecies), EzGenome and JSpecies provided very similar results. However, the ANI calculator provided higher intra- and inter-species values than the other two tools (differences within the ranges 0.06–0.82% and 0.92–3.38%, respectively). Nevertheless each of these tools produced the same species classification for the studied Aeromonas genomes. To avoid possible misinterpretations with the ANI calculator, particularly when values are at the borderline of the 95% cutoff, one of the other calculation tools (EzGenome or JSpecies) should be used in combination. It is recommended that once a genome sequence is obtained the correct taxonomic affiliation is verified using ANI or a MLPA before it is submitted to the NCBI and that researchers should amend the existing taxonomic errors present in databases.
doi:10.1371/journal.pone.0115813
PMCID: PMC4301921  PMID: 25607802
3.  A Hypothetical Protein of Alteromonas macleodii AltDE1 (amad1_06475) Predicted to be a Cold-Shock Protein with RNA Chaperone Activity 
Alteromonas macleodii AltDE1 is a deep sea protobacteria that is distinct from the surface isolates of the same species. This study was designed to elucidate the biological function of amad1_06475, a hypothetical protein of A. macleodii AltDE1. The 70 residues protein sequence showed considerable homology with cold-shock proteins (CSPs) and RNA chaperones from different organisms. Multiple sequence alignment further supported the presence of conserved csp domain on the protein sequence. The three-dimensional structure of the protein was also determined, and verified by PROCHECK, Verify3D, and QMEAN programs. The predicted structure contained five anti-parallel β-strands and RNA-binding motifs, which are characteristic features of prokaryotic CSPs. Finally, the binding of a thymidine-rich oligonucleotide and a single uracil molecule in the active site of the protein further strengthens our prediction about the function of amad1_06475 as a CSP and thereby acting as a RNA chaperone. The binding was performed by molecular docking tools and was compared with similar binding of 3PF5 (PDB) and 2HAX (PDB), major CSPs of Bacillus subtilis and Bacillus caldolyticus, respectively.
doi:10.4137/GRSB.S20802
PMCID: PMC4271719  PMID: 25574135
Cold-shock protein (CSP); RNA chaperone; cold-shock domain; RNP
4.  Aspect of Thrombolytic Therapy: A Review 
The Scientific World Journal  2014;2014:586510.
Thrombolytic therapy, also known as clot busting drug, is a breakthrough treatment which has saved untold lives. It has been used in the clinical area to treat venous and arterial thromboembolic complaints which are a foremost cause of death. In 1761, Morgagni lead the way of thrombolytic therapy. Now day's different types of thrombolytic drugs are currently available in market: alteplase, anistreplase, urokinase, streptokinase, tenecteplase, and so forth. Thrombolytic therapy should be given with maintaining proper care in order to minimize the risk of clinically important bleeding as well as enhance the chances of successfully thrombolysis of clot. These cares include preinfusion care, during the infusion care, and postinfusion care. Besides proper knowledge of contraindication, evolutionary factor, and combination of drug is essential for successful thrombolytic therapy. In these review we discussed about these aspect of thrombolytic therapy.
doi:10.1155/2014/586510
PMCID: PMC4276353  PMID: 25574487
5.  Genome-wide Analysis of Mycoplasma hominis for the Identification of Putative Therapeutic Targets 
Drug Target Insights  2014;8:51-62.
Ever increasing propensity of antibiotic resistance among pathogenic bacteria raises the demand for the development of novel therapeutic agents to control this grave problem. Advances in the field of bioinformatics, genomics, and proteomics have greatly facilitated the discovery of alternative drugs by swift identification of new drug targets. In the present study, we employed comparative genomics and metabolic pathway analysis with an aim of identifying therapeutic targets in Mycoplasma hominis. Our study has revealed 40 annotated metabolic pathways, including five unique pathways of M. hominis. Our study also identified 179 essential proteins, including 59 proteins having no similarity with human proteins. Further filtering by molecular weight, subcellular localization, functional analysis, and protein network interaction, we identified 57 putative candidates for which new drugs can be developed. Druggability analysis for each of the identified targets has prioritized 16 proteins as suitable for potential drug development.
doi:10.4137/DTI.S19728
PMCID: PMC4263438  PMID: 25574133
Mycoplasma hominis; drug targets; bioinformatics; genomics; proteomics
6.  Taxonomic Affiliation of New Genomes Should Be Verified Using Average Nucleotide Identity and Multilocus Phylogenetic Analysis 
Genome Announcements  2014;2(6):e00927-14.
The average nucleotide identity (ANI) determines if two genomes belong to the same species. Using ANI, we detected mislabeled genomes and recommend verifying with ANI and multilocus phylogenetic analysis the species affiliations of the announced genomes. The slightly different results obtained with different ANI calculation software can potentially mislead taxonomic inferences.
doi:10.1128/genomeA.00927-14
PMCID: PMC4256179  PMID: 25477398
7.  Molecular Characterization of Legionellosis Drug Target Candidate Enzyme Phosphoglucosamine Mutase from Legionella pneumophila (strain Paris): An In Silico Approach 
Genomics & Informatics  2014;12(4):268-275.
The harshness of legionellosis differs from mild Pontiac fever to potentially fatal Legionnaire's disease. The increasing development of drug resistance against legionellosis has led to explore new novel drug targets. It has been found that phosphoglucosamine mutase, phosphomannomutase, and phosphoglyceromutase enzymes can be used as the most probable therapeutic drug targets through extensive data mining. Phosphoglucosamine mutase is involved in amino sugar and nucleotide sugar metabolism. The purpose of this study was to predict the potential target of that specific drug. For this, the 3D structure of phosphoglucosamine mutase of Legionella pneumophila (strain Paris) was determined by means of homology modeling through Phyre2 and refined by ModRefiner. Then, the designed model was evaluated with a structure validation program, for instance, PROCHECK, ERRAT, Verify3D, and QMEAN, for further structural analysis. Secondary structural features were determined through self-optimized prediction method with alignment (SOPMA) and interacting networks by STRING. Consequently, we performed molecular docking studies. The analytical result of PROCHECK showed that 95.0% of the residues are in the most favored region, 4.50% are in the additional allowed region and 0.50% are in the generously allowed region of the Ramachandran plot. Verify3D graph value indicates a score of 0.71 and 89.791, 1.11 for ERRAT and QMEAN respectively. Arg419, Thr414, Ser412, and Thr9 were found to dock the substrate for the most favorable binding of S-mercaptocysteine. However, these findings from this current study will pave the way for further extensive investigation of this enzyme in wet lab experiments and in that way assist drug design against legionellosis.
doi:10.5808/GI.2014.12.4.268
PMCID: PMC4330265
docking analysis; drug delivery systems; homology modeling; Legionella pneumophila; legionellosis
8.  Early Depletion of Primordial Germ Cells in Zebrafish Promotes Testis Formation 
Stem Cell Reports  2014;4(1):61-73.
Summary
As complete absence of germ cells leads to sterile males in zebrafish, we explored the relationship between primordial germ cell (PGC) number and sexual development. Our results revealed dimorphic proliferation of PGCs in the early zebrafish larvae, marking the beginning of sexual differentiation. We applied morpholino-based gene knockdown and cell transplantation strategies to demonstrate that a threshold number of PGCs is required for the stability of ovarian fate. Using histology and transcriptomic analyses, we determined that zebrafish gonads are in a meiotic ovarian stage at 14 days postfertilization and identified signaling pathways supporting meiotic oocyte differentiation and eventual female fate. The development of PGC-depleted gonads appears to be restrained and delayed, suggesting that PGC number may directly regulate the variability and length of gonadal transformation and testicular differentiation in zebrafish. We propose that gonadal transformation may function as a developmental buffering mechanism to ensure the reproductive outcome.
Highlights
•A dimorphic proliferation of germ cells occurs in early zebrafish•A threshold number of PGCs is required for the ovarian fate•Germ cell numbers may regulate the variability of gonadal transformation•Gonadal transformation may function as a buffering mechanism for sexual development
In this article, Orban, Tzung, and colleagues report that the number of primordial germ cells (PGCs) plays a regulatory role during sexual differentiation in zebrafish. Partial depletion of PGCs leads to male bias, and a threshold number is required for ovarian development. They further provide evidence suggesting that the PGC-depleted gonads might undergo a less pronounced gonadal transformation than WTs.
doi:10.1016/j.stemcr.2014.10.011
PMCID: PMC4297871  PMID: 25434820
9.  Draft Genome Sequences of Two Novel Aeromonas Species Recovered in Association with Cyanobacterial Blooms 
Genome Announcements  2014;2(6):e01181-14.
Aeromonas aquatica and Aeromonas lacus are two new species that have been found in association with cyanobacterial blooms from recreational Finnish lakes where adverse human health effects have been recorded. Here, we present the draft genome sequences of their type strains.
doi:10.1128/genomeA.01181-14
PMCID: PMC4239355  PMID: 25414500
10.  In vivo antipyretic, antiemetic, in vitro membrane stabilization, antimicrobial, and cytotoxic activities of different extracts from Spilanthes paniculata leaves 
Biological Research  2014;47(1):45.
Background
The study was conducted to evaluate the in vitro antimicrobial activity, cytotoxic, and membrane stabilization activities, and in vivo antiemetic and antipyretic potentials of ethanolic extract, n-hexane and ethyl acetate soluble fractions of Spilanthes paniculata leaves for the first time widely used in the traditional treatments in Bangladesh.
Results
In antipyretic activity assay, a significant reduction (P < 0.05) was observed in the temperature in the mice tested. At dose 400 mg/kg-body weight, the n-hexane soluble fraction showed the effect (36.7 ± 0.63°C ) as like as the standard (dose 150 mg/kg-body weight) after 5 h of administration. Extracts showed significant (P < 0.001) potential when tested for the antiemetic activity compared to the standard, metoclopramide. At dose 50 mg/kg-body weight, the standard showed 67.23% inhibition, whereas n-hexane and ethyl acetate soluble fractions showed 37.53% and 24.93% inhibition of emesis respectively at dose 400 mg/kg-body weight. In antimicrobial activity assay, the n-hexane soluble fraction (400 μg/disc) showed salient activity against the tested organisms. It exerts highest activity against Salmonella typhi (16.9 mm zone of inhibition); besides, crude, and ethyl acetate extracts showed resistance to Bacillus cereus and Bacillus subtilis, and Vibrio cholera respectively. All the extracts were tested for lysis of the erythrocytes. At the concentration of 1mg/ml, ethanol extract, and n-hexane and ethyl acetate soluble fractions significantly inhibited hypotonic solution induced lysis of the human red blood cell (HRBC) (27.406 ± 3.57, 46.034 ± 3.251, and 30.72 ± 5.679% respectively); where standard drug acetylsalicylic acid (concentration 0.1 mg/ml) showed 77.276 ± 0.321% inhibition. In case of heat induced HRBC hemolysis, the plant extracts also showed significant activity (34.21 ± 4.72, 21.81 ± 3.08, and 27.62 ± 8.79% inhibition respectively). In the brine shrimp lethality bioassay, the n-hexane fraction showed potent (LC50 value 48.978 μg/ml) activity, whereas ethyl acetate fraction showed mild (LC50 value 216.77 μg/ml) cytotoxic activity.
Conclusions
Our results showed that the n-hexane extract has better effects than the other in all trials. In the context, it can be said that the leaves of S. paniculata possess remarkable pharmacological effects, and justify its folkloric use as antimicrobial, antipyretic, anti-inflammatory, and antiemetic agent. Therefore, further research may be suggested to find possible mode of action of the plant part.
doi:10.1186/0717-6287-47-45
PMCID: PMC4177068  PMID: 25299748
Antiemetic; Anti-inflammatory; Antimicrobial activity; Brine shrimp lethality bioassay; Cytotoxic; Membrane stabilization; Spilanthes paniculata
11.  Phytochemical, Phytotherapeutical and Pharmacological Study of Momordica dioica 
Momordica dioica is a perennial, dioecious, cucurbitaceous climbing creeper (commonly known as kakrol, spiny gourd or teasle gourd). It is native to Asia with extensive distribution in India and Bangladesh. It is used not only as preventive and curative agent for various diseases but also as vegetable with a significant nutritional value over thousands of years. This review aims to take an attempt to evaluate the phytochemical, ethnobotanical, phytotherapeutical and pharmacological properties of kakrol according to the view of traditional medicinal plant based treatment including ayurveda along with recent scientific observations. Kakrol is considered as an underutilized vegetable, although having significant presence of certain compounds containing higher nutritional value than many frequently consumed vegetables. Moreover, as a traditional medicinal plant, it is still potential for its phytochemical components that increase the demand of further extensive evaluation to justify its other therapeutical roles. Therefore, this effort will be helpful to researchers who interested to disclose the unjustified phytotherapeutical role of Momordica dioica.
doi:10.1155/2014/806082
PMCID: PMC4145798  PMID: 25197312
12.  Mental disorders in Bangladesh: a systematic review 
BMC Psychiatry  2014;14(1):216.
Background
Mental disorders constitute a major public health problem globally with higher burden in low and middle-income countries. In Bangladesh, systematically-collected data on mental disorders are scarce and this leaves the extent of the problem not so well defined. We reviewed the literature on mental health disorders in Bangladesh to summarize the available data and identify evidence gaps.
Methods
We identified relevant literature on mental disorders within Bangladesh published between 1975 and October, 2013 through a systematic and comprehensive search. Relevant information from the selected articles was extracted and presented in tables.
Results
We identified 32 articles which met our pre-defined eligibility criteria. The reported prevalence of mental disorders varied from 6.5 to 31.0% among adults and from 13.4 to 22.9% among children. Some awareness regarding mental health disorders exists at community level. There is a negative attitude towards treatment of those affected and treatment is not a priority in health care delivery. Mental health services are concentrated around tertiary care hospitals in big cities and absent in primary care.
Conclusions
The burden of mental disorders is high in Bangladesh, yet a largely unrecognized and under-researched area. To improve the mental health services in Bangladesh, further well-designed epidemiological and clinical research are needed.
doi:10.1186/s12888-014-0216-9
PMCID: PMC4149198  PMID: 25073970
Mental disorders; Depression; Systematic review; Prevalence; Comorbidity; Service delivery; Management; Treatment; Bangladesh
13.  Fluoroquinolone Resistance Mechanisms of Shigella flexneri Isolated in Bangladesh 
PLoS ONE  2014;9(7):e102533.
Objective
To investigate the prevalence and mechanisms of fluoroquinolone resistance in Shigella species isolated in Bangladesh and to compare with similar strains isolated in China.
Methods
A total of 3789 Shigella isolates collected from Clinical Microbiology Laboratory of icddr,b, during 2004–2010 were analyzed for antibiotic susceptibility. Analysis of plasmids, plasmid-mediated quinolone-resistance genes, PFGE, and sequencing of genes of the quinolone-resistance-determining regions (QRDR) were conducted in representative strains isolated in Bangladesh and compared with strains isolated in Zhengding, China. In addition, the role of efflux-pump was studied by using the efflux-pump inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP).
Results
Resistance to ciprofloxacin in Shigella species increased from 0% in 2004 to 44% in 2010 and S. flexneri was the predominant species. Of Shigella spp, ciprofloxacin resistant (CipR) strains were mostly found among S. flexneri (8.3%), followed by S. sonnei (1.5%). Within S. flexneri (n = 2181), 14.5% were resistance to ciprofloxacin of which serotype 2a was predominant (96%). MIC of ciprofloxacin, norfloxacin, and ofloxacin were 6–32 mg/L, 8–32 mg/L, and 8–24 mg/L, respectively in S. flexneri 2a isolates. Sequencing of QRDR genes of resistant isolates showed double mutations in gyrA gene (Ser83Leu, Asp87Asn/Gly) and single mutation in parC gene (Ser80Ile). A difference in amino acid substitution at position 87 was found between strains isolated in Bangladesh (Asp87Asn) and China (Asp87Gly) except for one. A novel mutation at position 211 (His→Tyr) in gyrA gene was detected only in the Bangladeshi strains. Susceptibility to ciprofloxacin was increased by the presence of CCCP indicating the involvement of energy dependent active efflux pumps. A single PFGE type was found in isolates from Bangladesh and China suggesting their genetic relatedness.
Conclusions
Emergence of fluoroquinolone resistance in Shigella undermines a major challenge in current treatment strategies which needs to be followed up by using empirical therapeutic strategies.
doi:10.1371/journal.pone.0102533
PMCID: PMC4100904  PMID: 25028972
14.  Is Peripheral Immunity Regulated by Blood-Brain Barrier Permeability Changes? 
PLoS ONE  2014;9(7):e101477.
S100B is a reporter of blood-brain barrier (BBB) integrity which appears in blood when the BBB is breached. Circulating S100B derives from either extracranial sources or release into circulation by normal fluctuations in BBB integrity or pathologic BBB disruption (BBBD). Elevated S100B matches the clinical presence of indices of BBBD (gadolinium enhancement or albumin coefficient). After repeated sub-concussive episodes, serum S100B triggers an antigen-driven production of anti-S100B autoantibodies. We tested the hypothesis that the presence of S100B in extracranial tissue is due to peripheral cellular uptake of serum S100B by antigen presenting cells, which may induce the production of auto antibodies against S100B. To test this hypothesis, we used animal models of seizures, enrolled patients undergoing repeated BBBD, and collected serum samples from epileptic patients. We employed a broad array of techniques, including immunohistochemistry, RNA analysis, tracer injection and serum analysis. mRNA for S100B was segregated to barrier organs (testis, kidney and brain) but S100B protein was detected in immunocompetent cells in spleen, thymus and lymph nodes, in resident immune cells (Langerhans, satellite cells in heart muscle, etc.) and BBB endothelium. Uptake of labeled S100B by rat spleen CD4+ or CD8+ and CD86+ dendritic cells was exacerbated by pilocarpine-induced status epilepticus which is accompanied by BBBD. Clinical seizures were preceded by a surge of serum S100B. In patients undergoing repeated therapeutic BBBD, an autoimmune response against S100B was measured. In addition to its role in the central nervous system and its diagnostic value as a BBBD reporter, S100B may integrate blood-brain barrier disruption to the control of systemic immunity by a mechanism involving the activation of immune cells. We propose a scenario where extravasated S100B may trigger a pathologic autoimmune reaction linking systemic and CNS immune responses.
doi:10.1371/journal.pone.0101477
PMCID: PMC4079719  PMID: 24988410
15.  Diagnosed hematological malignancies in Bangladesh - a retrospective analysis of over 5000 cases from 10 specialized hospitals 
BMC Cancer  2014;14:438.
Background
The global burden from cancer is rising, especially as low-income countries like Bangladesh observe rapid aging. So far, there are no comprehensive descriptions reporting diagnosed cancer group that include hematological malignancies in Bangladesh.
Methods
This was a multi-center hospital-based retrospective descriptive study of over 5000 confirmed hematological cancer cases in between January 2008 to December 2012. Morphological typing was carried out using the “French American British” classification system.
Results
A total of 5013 patients aged between 2 to 90 years had been diagnosed with malignant hematological disorders. A 69.2% were males (n = 3468) and 30.8% females (n = 1545), with a male to female ratio of 2.2:1. The overall median age at diagnosis was 42 years. Acute myeloid leukemia was most frequent (28.3%) with a median age of 35 years, followed by chronic myeloid leukemia with 18.2% (median age 40 years), non-Hodgkin lymphoma (16.9%; median age 48 years), acute lymphoblastic leukemia (14.1%; median age 27 years), multiple myeloma (10.5%; median age 55 years), myelodysplastic syndromes (4.5%; median age 57 years) and Hodgkin’s lymphoma (3.9%; median age 36 years). The least common was chronic lymphocytic leukemia (3.7%; median age 60 years). Below the age of 20 years, acute lymphoblastic leukemia was predominant (37.3%), followed by acute myeloid leukemia (34%). Chronic lymphocytic leukemia and multiple myeloma had mostly occurred among older patients, aged 50-over.
Conclusions
For the first time, our study presents the pattern and distribution of diagnosed hematological cancers in Bangladesh. It shows differences in population distributions as compared to other settings with possibly a lower presence of non-Hodgkin lymphoma. There might be under-reporting of affected women. Further studies are necessary on the epidemiology, genetics and potential environmental risk factors within this rapidly aging country.
doi:10.1186/1471-2407-14-438
PMCID: PMC4063230  PMID: 24929433
AML; CML; ALL; MDS; NHL; HL; MM; Hematological malignancy; Bangladesh
16.  An Asian Origin of Virulent Aeromonas hydrophila Responsible for Disease Epidemics in United States-Farmed Catfish 
mBio  2014;5(3):e00848-14.
ABSTRACT
Since 2009, catfish farming in the southeastern United States has been severely impacted by a highly virulent and clonal population of Aeromonas hydrophila causing motile Aeromonas septicemia (MAS) in catfish. The possible origin of this newly emerged highly virulent A. hydrophila strain is unknown. In this study, we show using whole-genome sequencing and comparative genomics that A. hydrophila isolates from diseased grass carp in China and catfish in the United States have highly similar genomes. Our phylogenomic analyses suggest that U.S. catfish isolates emerged from A. hydrophila populations of Asian origin. Furthermore, we identified an A. hydrophila strain isolated in 2004 from a diseased catfish in Mississippi, prior to the onset of the major epidemic outbreaks in Alabama starting in 2009, with genomic characteristics that are intermediate between those of the Asian and Alabama fish isolates. Investigation of A. hydrophila strain virulence demonstrated that the isolate from the U.S. catfish epidemic is significantly more virulent to both channel catfish and grass carp than is the Chinese carp isolate. This study implicates the importation of fish or fishery products into the United States as the source of highly virulent A. hydrophila that has caused severe epidemic outbreaks in United States-farmed catfish and further demonstrates the potential for invasive animal species to disseminate bacterial pathogens worldwide.
IMPORTANCE
Catfish aquaculture farming in the southeastern United States has been severely affected by the emergence of virulent Aeromonas hydrophila responsible for epidemic disease outbreaks, resulting in the death of over 10 million pounds of catfish. Because the origin of this newly emerged A. hydrophila strain is unknown, this study used a comparative genomics approach to conduct a phylogenomic analysis of A. hydrophila isolates obtained from the United States and Asia. Our results suggest that the virulent isolates from United States-farmed catfish have a recent common ancestor with A. hydrophila isolates from diseased Asian carp. We have also observed that an Asian carp isolate, like recent U.S. catfish isolates, is virulent in catfish. The results from this study suggest that the highly virulent U.S. epidemic isolates emerged from an Asian source and provide another example of the threat that invasive species pose in the dissemination of bacterial pathogens.
doi:10.1128/mBio.00848-14
PMCID: PMC4049099  PMID: 24895303
17.  Recombinant TLR5 Agonist CBLB502 Promotes NK Cell-Mediated Anti-CMV Immunity in Mice 
PLoS ONE  2014;9(5):e96165.
Prior work using allogeneic bone marrow transplantation (allo-BMT) models showed that peritransplant administration of flagellin, a toll-like receptor 5 (TLR5) agonist protected murine allo-BMT recipients from CMV infection while limiting graft-vs-host disease (GvHD). However, the mechanism by which flagellin-TLR5 interaction promotes anti-CMV immunity was not defined. Here, we investigated the anti-CMV immunity of NK cells in C57BL/6 (B6) mice treated with a highly purified cGMP grade recombinant flagellin variant CBLB502 (rflagellin) followed by murine CMV (mCMV) infection. A single dose of rflagellin administered to mice between 48 to 72 hours prior to MCMV infection resulted in optimal protection from mCMV lethality. Anti-mCMV immunity in rflagellin-treated mice correlated with a significantly reduced liver viral load and increased numbers of Ly49H+ and Ly49D+ activated cytotoxic NK cells. Additionally, the increased anti-mCMV immunity of NK cells was directly correlated with increased numbers of IFN-γ, granzyme B- and CD107a producing NK cells following mCMV infection. rFlagellin-induced anti-mCMV immunity was TLR5-dependent as rflagellin-treated TLR5 KO mice had ∼10-fold increased liver viral load compared with rflagellin-treated WT B6 mice. However, the increased anti-mCMV immunity of NK cells in rflagellin-treated mice is regulated indirectly as mouse NK cells do not express TLR5. Collectively, these data suggest that rflagellin treatment indirectly leads to activation of NK cells, which may be an important adjunct benefit of administering rflagellin in allo-BMT recipients.
doi:10.1371/journal.pone.0096165
PMCID: PMC4039429  PMID: 24879439
18.  Isolation, fractionation and identification of chemical constituents from the leaves crude extracts of Mentha piperita L grown in Sultanate of Oman 
Objective
To analyze and identify the chemical compositions of different organic plants crude extracts of Mentha piperita (M. piperita) grown in Sultanate of Oman by gas chromatography-mass spectrometry (GC-MS).
Methods
The powder sample was extracted with methanol by using Soxhlet extractor. Methanol crude extracts of M. piperita and its derived fractions of hexane, chloroform, ethyl acetate and butanol were prepared.
Results
Qualitative analyses of various organic plant crude extracts of M. piperita by using GC-MS showed that majority of these compounds are bioactive.
Conclusions
According to the results of the present study, the plant crude extracts could be used as medicine for the treatment of different diseases. The analysis and identification of the chemical compounds in the plant crude extracts by using GC-MS was the first time.
doi:10.12980/APJTB.4.2014C1051
PMCID: PMC4025344  PMID: 25183113
Mentha piperita; Organic crude extracts; Soxhlet extractor; Gas chromatography-mass spectrometry analyses
19.  Anti-vascular endothelial growth factor therapy-induced glioma invasion is associated with accumulation of Tie2-expressing monocytes 
Oncotarget  2014;5(8):2208-2220.
The addition of anti-angiogenic therapy to the few treatments available to patients with malignant gliomas was based on the fact that these tumors are highly vascularized and on encouraging results from preclinical and clinical studies. However, tumors that initially respond to this therapy invariably recur with the acquisition of a highly aggressive and invasive phenotype. Although several myeloid populations have been associated to this pattern of recurrence, a specific targetable population has not been yet identified. Here, we present evidence for the accumulation of Tie2-expressing monocytes/macrophages (TEMs) at the tumor/normal brain interface of mice treated with anti-VEGF therapies in regions with heightened tumoral invasion. Furthermore, we describe the presence of TEMs in malignant glioma surgical specimens that recurred after bevacizumab treatment. Our studies showed that TEMs enhanced the invasive properties of glioma cells and secreted high levels of gelatinase enzymatic proteins. Accordingly, Tie2+MMP9+ monocytic cells were consistently detected in the invasive tumor edge upon anti-VEGF therapies. Our results suggest the presence of a specific myeloid/monocytic subpopulation that plays a pivotal role in the mechanism of escape of malignant gliomas from anti-VEGF therapies and therefore constitutes a new cellular target for combination therapies in patients selected for anti-angiogenesis treatment.
PMCID: PMC4039157  PMID: 24809734
Brain tumors; antiangiogenesis; Tie2; tumor microenvironment; monocyte
20.  The prevalence of naturally acquired swimming ability among children in Bangladesh: a cross sectional survey 
BMC Public Health  2014;14:404.
Background
Most rural homes in Bangladesh have ponds nearby to serve as household water sources. As a result children of all ages are exposed to water bodies on a daily basis. Children learn to swim early in childhood from peers and relatives in a natural process that involves play and structured learning. In a large, national injury survey in Bangladesh, the ability to swim was associated with reduced risk of drowning. This study determines the prevalence of swimming ability in children in Bangladesh as a step in assessing whether this is a potential component of a national drowning prevention program.
Methods
A descriptive study design using a subset of a national sample survey determined the prevalence of naturally acquired swimming ability (NASA) reported by children of rural and urban communities in Bangladesh. A total of 2,598 households (1,999 rural and 599 urban) housing 4,336 children (2,263 male and 2,073 female) aged 5-17 years were chosen from 4 randomly selected districts using multistage random sampling. NASA was defined as the ability to cross 25 meters of water deeper than the child’s height using any body movement for self-propulsion.
Results
Reported NASA was greater in males (55.6%) than females (47.9%) and among rural children (57.8%) than urban children (25.5%) for children 5-17 years. The proportion reporting NASA increased with increasing age. At age 5, 5.8% of males and 6.3% of females reported NASA, rising to 84.3% of males and 70.7% of females by age 17. By age 17, 83.1% of rural children and 57.5% of urban children reported NASA.
Conclusion
Most children in Bangladesh report being able to swim 25 meters and learning it by middle childhood. Reported NASA is higher for males than females and for rural children than urban children. High rates of swimming appear to be achievable in the absence of pools and a swim-teaching industry. This may facilitate development of a low cost, national drowning prevention program with swimming an integral part.
doi:10.1186/1471-2458-14-404
PMCID: PMC4014625  PMID: 24767407
Swimming ability; Naturally acquired swimming ability; NASA; Child drowning; Drowning prevention; Bangladesh
21.  Soluble FAS ligand as a biomarker of disease recurrence in differentiated thyroid cancer 
Cancer  2013;119(8):1503-1511.
Background
Reliable predictive biomarkers are required to address the challenge of disease recurrence following thyroid cancer surgery. We assessed the association of cellular- and serum-based immunologic mediators with thyroid cancer recurrence.
Methods
Leukocyte subset counts and immune regulatory cytokines were determined in peripheral blood samples using multiparameter flow cytometry and 51-panel multiplex ELISA (Luminex) assays, respectively. Functional activity of circulating B-, T- and NK lymphocytes was assessed ex vivo. Differences in mean biomarker levels between defined subject groups and correlations between biomarkers and cancer recurrence were assessed by t-test or Wilcoxon test and by univariate and multivariate analyses with Cox model. Optimal cut-off values of significantly correlated biomarkers that best predict recurrence post surgery were established by receiver operating characteristics and validated by the optimal cut-point determination algorithm.
Results
We enrolled 35 patients (median age - 49.4 years; 24 females; 15 patients with disease recurrence) and 21 controls. Patients without recurrence had higher levels of soluble FAS ligand (sFASL), TGF-b, T-regs and PD-1/PD-L1 expressing leukocytes. sFASL (HR:0.60; 95%CI:0.38–0.95; p=0.031) and IFN-α (HR:1.55; 95%CI:1.03–2.34; p=0.038) showed significant association with cancer recurrence. There was a significant difference in PFS between patient groups stratified by sFASL optimal cut-point of 15 pg/ml (Logrank p=0.0009).
Conclusions
sFASL and IFN-α showed significant correlation with thyroid cancer recurrenceand may be useful for risk-adapted surveillance strategies for thyroid cancer.
doi:10.1002/cncr.27937
PMCID: PMC3799834  PMID: 23340930
Thyroid; cancer; biomarker; recurrence; FASL
22.  Development of puffed ginseng-rice snack from ginseng powder and map rice flour using steam and compression process 
A new manufacturing method for producing a puffed ginseng-rice snack (PGRS) was developed using ginseng powder and map rice flour through a steam and compression process (SCP). The physical and sensory properties of the PGRS were characterized. The pellets for puffing were prepared from ginseng powder and map rice flour. The pellets were subjected to 16, 18, and 20% moisture contents and were puffed at 225, 235, and 245° C. The specific volumes of the PGRSs increased with heating temperature and moisture content. However, the breaking strength of the PGRSs decreased. In addition, the SCP imposed special features in the PGRSs that made them more acceptable. The Hunter L-value increased with heating temperature and moisture content. These results indicate that a PGRS with functional additives could be effectively developed into a functional food with the use of a puffing machine, and that the PGRS shows potential as a new snack product.
doi:10.1007/s13197-012-0626-y
PMCID: PMC3550922  PMID: 24425934
Puffed ginseng-rice snack; Steam; Compression; Map rice
23.  Conventional or Interpersonal Communication: Which Works Best in Disseminating Malaria Information in an Endemic Rural Bangladeshi Community? 
PLoS ONE  2014;9(3):e90711.
Background
Since 2007, BRAC has been implementing malaria prevention and control programme in 13 endemic districts of Bangladesh under the National Malaria Control Programme. This study was done to examine the role of different communication media in bringing about changes in knowledge and awareness which facilitate informed decision-making for managing malaria-like illnesses.
Methods
A baseline survey in 2007 before inception of the programme, and a follow-up survey in 2012 were done to study changes in different aspects of programme interventions including the communication component. Both the surveys used the same sampling technique to select 25 households at random from each of the 30 mauza/villages in a district. A pre-tested, semi-structured questionnaire was used to collect relevant information from respondents in face-to-face interview. Analysis was done comparing the study areas at two different times. Statistical tests were done as necessary to examine the differences.
Results
The intervention succeeded in improving knowledge in some trivial areas (e.g., most frequent symptom suggestive of malaria, importance of using insecticidal bed nets) but not in critical domains necessary for taking informed action (e.g., mode of malaria transmission, awareness about facilities providing free malaria treatment). Inequity in knowledge and practice was quite common depending upon household affluence, location of households in high or low endemic districts, and sex. Of the different media used in Information, Education and communication (IEC) campaigns during the study period, interpersonal communication with community health workers/relatives/neighbours/friends was found to be more effective in improving knowledge and practice than conventional print and audio-visual media.
Conclusion
This study reiterates the fact that conventional media may not be user-friendly or culture-sensitive for this semi-literate/illiterate community where dissemination through ‘words of mouth’ is more common, and as such, interpersonal communication is more effective. This is especially important for initiating informed action by the community in managing malaria-like illnesses.
doi:10.1371/journal.pone.0090711
PMCID: PMC3948336  PMID: 24603849
24.  Prevalence of Iron-deficiency Anaemia among University Students in Noakhali Region, Bangladesh 
Iron-deficiency anaemia (IDA) is a common health problem in rural women and young children of Bangladesh. The university students usually take food from residential halls, and the food value of their diets is not always balanced. This cross-sectional study was conducted to estimate the prevalence of iron-deficiency anaemia among the university students of Noakhali region, Bangladesh. Haemoglobin level of 300 randomly-selected students was measured calorimetrically, using Sahli's haemoglobinometer during October to December 2011. Statistical analysis was done by using SPSS software for Windows (version 16) (SPSS Inc., Chicago, IL, USA). In the study, 55.3% students were found anaemic, of whom 36.7% were male, and 63.3% were female. Students aged 20-22 years were more anaemic (43.4%) than other age-groups. Majority (51.3%) of male students showed their haemoglobin level in the range of 13-15 g/dL, followed by 26.0% and 21.3% with 10-12 g/dL and 16-18 g/dL respectively. Although 50.5% anaemic and 51.1% non-anaemic female students showed normal BMI—lower percentage than anaemic (60.7%) and non-anaemic (71.9%) male students, the underweight students were found more anaemic than the overweight and obese subjects. Regular breakfast-taking habit showed significant (p=0.035, 95% CI 0.5-1.0) influence on IDA compared to non-regular breakfast takers. Consumption of meat, fish, poultry, eggs, or peanut butter regularly; junk food; multivitamins; and iron/iron-rich food showed insignificant (p=0.097, 95% CI 0.5-1.1; p=0.053, 95% CI 1.1-2.3; p=0.148, 95% CI 0.6-1.2; and p=0.487, 95% CI 0.7-1.4 respectively) role in provoking IDA. In the case of non-anaemic subjects, all of the above parameters were significant, except the junk food consumption (p=0.342, 95% CI 0.5-1.2). Our study revealed that majority of university students, especially female, were anaemic that might be aggravated by food habit and lack of awareness. The results suggest that anaemia can be prevented by providing proper knowledge on the healthful diet, improved lifestyle, and harmful effect of anaemia to the students.
PMCID: PMC4089078  PMID: 24847599
Iron-deficiency anaemia; Prevalence; University students; Bangladesh
25.  Infrastructure and Contamination of the Physical Environment in Three Bangladeshi Hospitals: Putting Infection Control into Context 
PLoS ONE  2014;9(2):e89085.
Objective
This paper describes the physical structure and environmental contamination in selected hospital wards in three government hospitals in Bangladesh.
Methods
The qualitative research team conducted 48 hours of observation in six wards from three Bangladeshi tertiary hospitals in 2007. They recorded environmental contamination with body secretions and excretions and medical waste and observed ward occupant handwashing and use of personal protective equipment. They recorded number of persons, number of open doors and windows, and use of fans. They measured the ward area and informally observed waste disposal outside the wards. They conducted nine focus group discussions with doctors, nurses and support staff.
Results
A median of 3.7 persons were present per 10 m2 of floor space in the wards. A median of 4.9 uncovered coughs or sneezes were recorded per 10 m2 per hour per ward. Floors in the wards were soiled with saliva, spit, mucous, vomitus, feces and blood 125 times in 48 hours. Only two of the 12 patient handwashing stations had running water and none had soap. No disinfection was observed before or after using medical instruments. Used medical supplies were often discarded in open containers under the beds. Handwashing with soap was observed in only 32 of 3,373 handwashing opportunities noted during 48 hours. Mosquitoes and feral cats were commonly observed in the wards.
Conclusions
The physical structure and environment of our study hospitals are conducive to the spread of infection to people in the wards. Low-cost interventions on hand hygiene and cleaning procedures for rooms and medical equipment should be developed and evaluated for their practicality and effectiveness.
doi:10.1371/journal.pone.0089085
PMCID: PMC3929649  PMID: 24586516

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