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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.
PMCID: PMC2807272  PMID: 19934601
calves; herniorrhaphy; reducible umbilical hernia
2.  An Asian Origin of Virulent Aeromonas hydrophila Responsible for Disease Epidemics in United States-Farmed Catfish 
mBio  2014;5(3):e00848-14.
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.
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.
PMCID: PMC4049099  PMID: 24895303
3.  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.
PMCID: PMC4039429  PMID: 24879439
4.  Isolation, fractionation and identification of chemical constituents from the leaves crude extracts of Mentha piperita L grown in Sultanate of Oman 
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).
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.
Qualitative analyses of various organic plant crude extracts of M. piperita by using GC-MS showed that majority of these compounds are bioactive.
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.
PMCID: PMC4025344
Mentha piperita; Organic crude extracts; Soxhlet extractor; Gas chromatography-mass spectrometry analyses
5.  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
6.  The prevalence of naturally acquired swimming ability among children in Bangladesh: a cross sectional survey 
BMC Public Health  2014;14:404.
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.
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.
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.
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.
PMCID: PMC4014625  PMID: 24767407
Swimming ability; Naturally acquired swimming ability; NASA; Child drowning; Drowning prevention; Bangladesh
7.  Soluble FAS ligand as a biomarker of disease recurrence in differentiated thyroid cancer 
Cancer  2013;119(8):1503-1511.
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.
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.
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).
sFASL and IFN-α showed significant correlation with thyroid cancer recurrenceand may be useful for risk-adapted surveillance strategies for thyroid cancer.
PMCID: PMC3799834  PMID: 23340930
Thyroid; cancer; biomarker; recurrence; FASL
8.  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.
PMCID: PMC3550922  PMID: 24425934
Puffed ginseng-rice snack; Steam; Compression; Map rice
9.  Conventional or Interpersonal Communication: Which Works Best in Disseminating Malaria Information in an Endemic Rural Bangladeshi Community? 
PLoS ONE  2014;9(3):e90711.
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.
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.
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.
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.
PMCID: PMC3948336  PMID: 24603849
10.  Infrastructure and Contamination of the Physical Environment in Three Bangladeshi Hospitals: Putting Infection Control into Context 
PLoS ONE  2014;9(2):e89085.
This paper describes the physical structure and environmental contamination in selected hospital wards in three government hospitals in Bangladesh.
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.
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.
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.
PMCID: PMC3929649  PMID: 24586516
11.  Prevalence and Diversity of Avian Haematozoan Parasites in Wetlands of Bangladesh 
The parasites of genera Haemoproteus, Plasmodium, and Leucocytozoon are well-known avian haematozoa and can cause declined productivity and high mortality in wild birds. The objective of the study was to record the prevalence of haematozoan parasites in a wide range of wetland birds in Bangladesh. Six species of Haemoproteus, seven species of Plasmodium, one unidentified species of Leucocytozoon, and one unidentified microfilaria of the genus Paronchocerca were found. Data on the morphology, size, hosts, prevalence, and infection intensity of the parasites are provided. The overall prevalence among the birds was 29.5% (95 out of 322 birds). Of those, 13.2% (42 of 319) of birds were infected with Haemoproteus spp., 15.1% with Plasmodium spp. (48 of 319) and 0.6% with Leucocytozoon spp. (2 of 319). Two birds were positive for both Haemoproteus sp. and Plasmodium sp. A single resident bird, Ardeola grayii, was found positive for an unidentified microfilaria. Prevalence of infection varied significantly among different bird families. Wild birds of Bangladesh carry several types of haematozoan parasites. Further investigation with a larger sample size is necessary to estimate more accurately the prevalence of haematozoan parasites among wild birds as well as domestic ducks for better understanding of the disease ecology.
PMCID: PMC3918735  PMID: 24587896
12.  Prevalence and Correlates of Smokeless Tobacco Consumption among Married Women in Rural Bangladesh 
PLoS ONE  2014;9(1):e84470.
To estimate the prevalence and identify correlates of smokeless tobacco consumption among married rural women with a history of at least one pregnancy in Madaripur, Bangladesh.
Materials and Methods
We conducted a cross-sectional survey using an interviewer administered, pre-tested, semi-structured questionnaire. All women living in the study area, aged 18 years and above with at least one pregnancy in their lifetime, who were on the electoral roll and agreed to participate were included in the study. Information on socio-demographic characteristics and smokeless tobacco consumption was collected. Smokeless tobacco consumption was categorized as ‘Current’, ‘Ever but not current’ and ‘Never’. Associations between smokeless tobacco consumption and the explanatory variables were estimated using simple and multiple binary logistic regression.
8074 women participated (response rate 99.9%). The prevalence of ‘Current consumption’, ‘Ever consumption but not current’, and ‘Never consumption’ was 25%, 44% and 31%, respectively. The mean age at first use was 31.5 years. 87% of current consumers reported using either Shadapata or Hakimpuree Jarda. Current consumption was associated with age, level of education, religion, occupation, being an income earner, marital status, and age at first use of smokeless tobacco. After adjustment for demographic variables, current consumption was associated with being over 25 years of age, a lower level of education, being an income earner, being Muslim, and being divorced, separated or widowed.
The prevalence of smokeless tobacco consumption is high among rural women in Bangladesh and the age of onset is considerably older than that for smoking. Smokeless tobacco consumption is likely to be producing a considerable burden of non-communicable disease in Bangladesh. Smokeless tobacco control strategies should be implemented.
PMCID: PMC3885568  PMID: 24416234
13.  Changing Trends in the Prevalence of Shigella Species: Emergence of Multi-Drug Resistant Shigella sonnei Biotype g in Bangladesh 
PLoS ONE  2013;8(12):e82601.
Shigellosis, caused by Shigella species, is a major public health problem in Bangladesh. To determine the prevalence and distribution of different Shigella species, we analyzed 10,827 Shigella isolates from patients between 2001 and 2011. S. flexneri was the predominant species isolated throughout the period. However, the prevalence of S. flexneri decreased from 65.7% in 2001 to 47% in 2011, whereas the prevalence of S. sonnei increased from 7.2% in 2001 to 25% in 2011. S. boydii and S. dysenteriae accounted for 17.3% and 7.7% of the isolates respectively throughout the period. Of 200 randomly selected S. sonnei isolates for extensive characterization, biotype g strains were predominant (95%) followed by biotype a (5%). Resistance to commonly used antibiotics including trimethoprim-sulfamethoxazole, nalidixic acid, ciprofloxacin, mecillinam and ampicillin was 89.5%, 86.5%, 17%, 10.5%, and 9.5%, respectively. All isolates were susceptible to ceftriaxone, cefotaxime, ceftazidime and imipenem. Ninety-eight percent of the strains had integrons belonging to class 1, 2 or both. The class 1 integron contained only dfrA5 gene, whereas among class 2 integron, 16% contained dhfrAI-sat1-aadA1-orfX gene cassettes and 84% harbored dhfrA1-sat2 gene cassettes. Plasmids of ∼5, ∼1.8 and ∼1.4 MDa in size were found in 92% of the strains, whereas only 33% of the strains carried the 120 MDa plasmid. PFGE analysis showed that strains having different integron patterns belonged to different clusters. These results show a changing trend in the prevalence of Shigella species with the emergence of multidrug resistant S. sonnei. Although S. flexneri continues to be the predominant species albeit with reduced prevalence, S. sonnei has emerged as the second most prevalent species replacing the earlier dominance by S. boydii and S. dysenteriae in Bangladesh.
PMCID: PMC3867351  PMID: 24367527
14.  Implication of Lateral Genetic Transfer in the Emergence of Aeromonas hydrophila Isolates of Epidemic Outbreaks in Channel Catfish 
PLoS ONE  2013;8(11):e80943.
To investigate the molecular basis of the emergence of Aeromonas hydrophila responsible for an epidemic outbreak of motile aeromonad septicemia of catfish in the Southeastern United States, we sequenced 11 A. hydrophila isolates that includes five reference and six recent epidemic isolates. Comparative genomics revealed that recent epidemic A. hydrophila isolates are highly clonal, whereas reference isolates are greatly diverse. We identified 55 epidemic-associated genetic regions with 313 predicted genes that are present in epidemic isolates but absent from reference isolates and 35% of these regions are located within genomic islands, suggesting their acquisition through lateral gene transfer. The epidemic-associated regions encode predicted prophage elements, pathogenicity islands, metabolic islands, fitness islands and genes of unknown functions, and 34 of the genes encoded in these regions were predicted as virulence factors. We found two pilus biogenesis gene clusters encoded within predicted pathogenicity islands. A functional metabolic island that encodes a complete pathway for myo-inositol catabolism was evident by the ability of epidemic A. hydrophila isolates to use myo-inositol as a sole carbon source. Testing of A. hydrophila field isolates found a consistent correlation between myo-inositol utilization as a sole carbon source and the presence of an epidemic-specific genetic marker. All epidemic isolates and one reference isolate shared a novel O-antigen cluster. Altogether we identified four different O-antigen biosynthesis gene clusters within the 11 sequenced A. hydrophila genomes. Our study reveals new insights into the evolutionary changes that have resulted in the emergence of recent epidemic A. hydrophila strains.
PMCID: PMC3835674  PMID: 24278351
15.  Complete Genome Sequence of a Channel Catfish Epidemic Isolate, Aeromonas hydrophila Strain ML09-119 
Genome Announcements  2013;1(5):e00755-13.
Aeromonas hydrophila is a Gram-negative, rod-shaped, mesophilic bacterium that infects both aquatic poikilothermic animals and mammals, including humans. Here, we present the complete genome sequence of Aeromonas hydrophila strain ML09-119, which represents a clonal group of A. hydrophila isolates causing outbreaks of bacterial septicemia in channel catfish since 2009.
PMCID: PMC3778208  PMID: 24051325
16.  Complete Genome Sequence of Staphylococcus aureus Tager 104, a Sequence Type 49 Ancestor 
Genome Announcements  2013;1(5):e00706-13.
We report here the complete genome sequence of Staphylococcus aureus Tager 104, originally isolated from a cutaneous abscess in 1947 by Morris Tager. Sequence typing of the strain revealed its membership in sequence type 49 (ST49), a previously unknown multilocus sequence type (MLST) in clinical samples.
PMCID: PMC3772141  PMID: 24029757
17.  Role for the Conserved N-terminal Cysteines in the Anti-Chemokine Activities by the Chemokine-like Protein MC148R1 Encoded by Molluscum Contagiosum Virus 
Virology  2011;417(2):449-456.
Molluscum contagiosum poxvirus (MCV) type 1 and type 2 encode two chemokine-like proteins MC148R1 and MC148R2. It is believed that MC148R proteins function by blocking the inflammatory response. However, the mechanism of the proposed biological activities of MC148R proteins and the role of the additional C-terminal cysteines that do not exist in other chemokines are not understood. Here, we demonstrated in two different assay systems that His-tagged MC148R1 displaces the interaction between CXCL12α and CXCR4. The N-terminal cysteines but not the additional C-terminal cysteines modulate this displacement. His-tagged MC148R1 blocked both CXCL12α-mediated and MIP-1α-mediated chemotaxis. In contrast, MC148R2 blocked MIP-1α-mediated but not CXCL12α-mediated chemotaxis. Immunoprecipitation by antibodies to MC148R1 or CXCL12α followed by immunoblotting and detection by antibodies to the other protein demonstrated physical interaction of His-tagged CXCL12α and His-tagged MC148R1. Interaction with chemokines might mask the receptor interaction site resulting in decreased binding and impairment of the biological activities.
PMCID: PMC3760151  PMID: 21802105
Chemokines; Chemotaxis; CXCL12; MCV; HIV
18.  Study of total phenol, flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris 
To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol, flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar, Nizwa, Sultanate of Oman.
The leave sample was extracted with methanol and evaporated. Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities. The prepare hexane, chloroform, ethyl acetate, butanol and methanol crude extracts were used for their evaluation of total phenol, flavonoids contents and phytochemical screening study. The established conventional methods were used for quantitative determination of total phenol, flavonoids contents and phytochemical screening.
Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids, saponins and steroids compounds. The result for total phenol content was the highest in butanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract.
The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.
PMCID: PMC3757278  PMID: 23998010
Thymus vulgaris L.; Various crude extracts; Phytochemical screening; Soxhlet extractor; Total phenol; Flavonoids contents
19.  Screening Utility, Local Perceptions, and Care-seeking for Reported Jaundeesh among Respondents Lacking Signs of Icterus in Rural Bangladesh 
In rural Bangladesh, acute viral hepatitis presents a significant burden on the public-health system. As part of the formative work for a large epidemiologic study of hepatitis E in rural Bangladesh, we sought to identify local terms that could be used for population-based screening of acute viral hepatitis. Exploration of the local term jaundeesh for screening utility identified a high burden of reported jaundeesh among individuals without symptoms of icterus. Recognizing that local perceptions of illness may differ from biomedical definitions of disease, we also sought to characterize the perceived aetiology, care-seeking patterns, diagnostic symptoms, and treatments for reported jaundeesh in the absence of icteric symptoms to inform future population-based studies on reported morbidities. We conducted a cross-sectional survey among 1,441 randomly-selected subjects to identify the prevalence of reported jaundeesh and to test the validity of this local term to detect signs of icterus. To characterize the perceived aetiology and care-seeking patterns for jaundeesh among the majority of respondents, we conducted in-depth interviews with 100 respondents who self-reported jaundeesh but lacked clinical signs of icterus. To describe diagnostic symptoms and treatments, in-depth interviews were also performed with 25 kabirajs or traditional faith healers commonly visited for jaundeesh. Of the 1,441 randomly-selected participants, one-fourth (n=361) reported jaundeesh, with only a third (n=122) reporting yellow eyes or skin, representative of icterus; Jaundeesh had a positive predictive value of 34% for detection of yellow eyes or skin. Anicteric patients with reported jaundeesh perceived their illnesses to result from humoral imbalances, most commonly treated by amulets, ritual handwashing, and bathing with herbal medicines. Jaundeesh patients primarily sought folk and spiritual remedies from informal care providers, with only 19% visiting allopathic care providers. Although the local term jaundeesh appeared to have limited epidemiologic utility to screen for acute symptomatic viral hepatitis, this term described a syndrome perceived to occur frequently in this population. Future population-based studies conducting surveillance for acute hepatitis should use caution in the use and interpretation of self-reported jaundeesh. Further study of jaundeesh may provide insight into the appropriate public-health response to this syndrome.
PMCID: PMC3805887  PMID: 24288951
Ethnography; Hepatitis; HEV; Jaundice; Medical anthropology; Morbidity; Traditional healers; Bangladesh
20.  Serogroup W135 Meningococcal Disease, The Gambia, 2012 
Emerging Infectious Diseases  2013;19(9):1507-1515.
In 2012, an outbreak of Neisseria meningitidis serogroup W135 occurred in The Gambia. The attack rate was highest among young children. The associated risk factors were male sex, contact with meningitis patients, and difficult breathing. Enhanced surveillance facilitates early epidemic detection, and multiserogroup conjugate vaccine could reduce meningococcal epidemics in The Gambia.
PMCID: PMC3810914  PMID: 23965435
Neisseria meningitidis; serogroup W135; meningitis; the Gambia; epidemic; outbreak; bacteria
21.  Health Care Utilization and Attitudes Survey: Understanding Diarrheal Disease in Rural Gambia 
Diarrheal disease causes ∼1.34 million deaths per year among children under 5 years of age globally. We conducted a Health Care Utilization and Attitudes Survey of 1,012 primary caregivers of children aged 0–11, 12–23, and 24–59 months randomly selected from a Demographic Surveillance population in rural Gambia. Point prevalence of diarrhea was 7.7% (95% confidence interval [CI] = 6.1–9.8); 23.3% had diarrhea within the previous 2 weeks. Caregivers of 81.5% of children with diarrhea sought healthcare outside their home, but only 48.4% of them visited a health center. Only 17.0% (95% CI = 12.1–23.2) of children with diarrhea received oral rehydration solution (ORS) at home. Abbreviated surveys conducted on six occasions over the subsequent 2 years showed no change in prevalence or treatment-seeking behavior. Diarrhea remains a significant problem in rural young Gambian children. Encouraging care-seeking behavior at health centers and promoting ORS use can reduce mortality and morbidity in this population.
PMCID: PMC3748496  PMID: 23629926
22.  Molecular epidemiology of canine GM1 gangliosidosis in the Shiba Inu breed in Japan: relationship between regional prevalence and carrier frequency 
Canine GM1 gangliosidosis is a fatal disease in the Shiba Inu breed, which is one of the most popular traditional breeds in Japan and is maintained as a standard breed in many countries. Therefore, it is important to control and reduce the prevalence of GM1 gangliosidosis for maintaining the quality of this breed and to ensure supply of healthy dogs to prospective breeders and owners. This molecular epidemiological survey was performed to formulate an effective strategy for the control and prevention of this disease.
The survey was carried out among 590 clinically unaffected Shiba Inu dogs from the 8 districts of Japan, and a genotyping test was used to determine nation-wide and regional carrier frequencies. The number and native district of affected dogs identified in 16 years from 1997 to June 2013 were also surveyed retrospectively. Of the 590 dogs examined, 6 dogs (1.02%, 6/590) were carriers: 3 dogs (2.27%, 3/132) from the Kinki district and the other 3 dogs from the Hokkaido, Kanto, and Shikoku districts. The retrospective survey revealed 23 affected dogs, among which, 19 dogs (82.6%) were born within the last 7 years. Of the 23 affected dogs, 12 dogs (52.2%) were from the Kinki district. Pedigree analysis demonstrated that all the affected dogs and carriers with the pedigree information have a close blood relationship.
Our results showed that the current carrier frequency for GM1 gangliosidosis is on the average 1.02% in Japan and rather high in the Kinki district, which may be related to the high prevalence observed over the past 16 years in this region. This observation suggests that carrier dogs are distributed all over Japan; however, kennels in the Kinki district may face an increased risk of GM1 gangliosidosis. Therefore, for effective control and prevention of this disease, it is necessary to examine as many breeding dogs as possible from all regions of Japan, especially from kennels located in areas with high prevalence and carrier frequency.
PMCID: PMC3701567  PMID: 23819787
GM1 Gangliosidosis; Shiba Inu Dog; Molecular Epidemiology; Canine Inherited Disease
23.  Human and Methodological Sources of Variability in the Measurement of Urinary 8-Oxo-7,8-dihydro-2′-deoxyguanosine 
Antioxidants & Redox Signaling  2013;18(18):2377-2391.
Aims: Urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed ‘spot’ samples showed strong correlations with 24 h excretion (the ‘gold’ standard) of urinary 8-oxodG (rp 0.67–0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine- or SG-adjusted first morning samples are recommended. Antioxid. Redox Signal. 18, 2377–2391.
PMCID: PMC3671631  PMID: 23198723
24.  Shaping T Cell – B Cell Collaboration in the Response to Human Immunodeficiency Virus Type 1 Envelope Glycoprotein gp120 by Peptide Priming 
PLoS ONE  2013;8(6):e65748.
Prime-boost vaccination regimes have shown promise for obtaining protective immunity to HIV. Poorly understood mechanisms of cellular immunity could be responsible for improved humoral responses. Although CD4+ T-cell help promotes B-cell development, the relationship of CD4+ T-cell specificity to antibody specificity has not been systematically investigated. Here, protein and peptide-specific immune responses to HIV-1 gp120 were characterized in groups of ten mucosally immunized BALB/c mice. Protein and peptide reactivity of serum antibody was tested for correlation with cytokine secretion by splenocytes restimulated with individual gp120 peptides. Antibody titer for gp120 correlated poorly with the peptide-stimulated T-cell response. In contrast, titers for conformational epitopes, measured as crossreactivity or CD4-blocking, correlated with average interleukin-2 and interleukin-5 production in response to gp120 peptides. Antibodies specific for conformational epitopes and individual gp120 peptides typically correlated with T-cell responses to several peptides. In order to modify the specificity of immune responses, animals were primed with a gp120 peptide prior to immunization with protein. Priming induced distinct peptide-specific correlations of antibodies and T-cells. The majority of correlated antibodies were specific for the primed peptides or other peptides nearby in the gp120 sequence. These studies suggest that the dominant B-cell subsets recruit the dominant T-cell subsets and that T-B collaborations can be shaped by epitope-specific priming.
PMCID: PMC3679139  PMID: 23776539
25.  In vitro antioxidant, cytotoxic, thrombolytic activities and phytochemical evaluation of methanol extract of the A. philippense L. leaves 
To study the leaves of Adiantum philippense L. for their antioxidant, cytotoxicity and thrombolytic activities and to perform phytochemical evaluation.
In-vitro antioxidant activity of extract was studied using DPPH radical scavenging, reducing power, total phenol and total flavonoid content determination assays. The cytotoxic activity was determined using brine shrimp lethality bioassay, thrombolytic activity by clot disruption and phytochemical potential by qualitative analysis.
The antioxidant activity of the extracts was found promising. The reducing power of this crude extract increase with the increase of concentration; IC50 values of DPPH scavenging activity was (140.00±0.86) µg/mL as compared to ascorbic acid [IC50 (130.00±0.76) µg/mL]; Total phenol and total flavonoids content were (148.26±0.24) mg/mL and (163.06±0.56) mg/mL respectively. In cytotoxicity assay the LC50 values of the sample was (106.41±0.78) µg/mL where as for standard vincristin sulphate was (08.50±0.24) µg/mL as a positive control and the extract shows (12.86±1.02)% clot lytic whereas standard streptokinase shows (30.86±0.44% clot lytic activity in thrombolytic assay. The phytochemical evaluation indicates the presence of chemical constituents including carbohydrates, alkaloids, saponins, glycosides, flavonoids.
This study shows that the methanol extract of leaves of Adiantum philippense L. has bioactivity but further compound isolation is necessary to confirm the activities of individual compounds.
PMCID: PMC3644574  PMID: 23730559
Adiantum philippense L.; Antioxidant; Cytotoxicity; Phytochemical evaluation

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