ABSTRACT
In this study 269 swabs collected from 254 ovine foot lesions and 15 apparently healthy ovine feet were screened by PCR for the presence of major lameness causing foot pathogens viz. Treponema species, D. nodosus, F. necrophorum and T. pyogenes with the presumption that ovine foot lesion positive for Treponema species alone or in association with other three pathogens were categorized as contagious ovine digital dermatitis (CODD). While samples positive for D. nodosus alone or its combination with F. necrophorum and T. pyogenes were considered as footrot (FR) and samples in which F. necrophorum or T. pyogenes was found either alone or in combination were considered as interdigital dermatitis (ID). The overall occurrence of Treponema sp. in ovine foot lesions was 48.0%, and ranged from 33 to 58%. In Treponema positive samples D. nodosus, F. necrophorum and T. pyogenes were present in 34 (27.4%), 66 (54.4%) and 84 (68.5%) in contrast to Treponema negative samples in which these were present in 15 (11.1%), 20 (14.12%) and 17 (12.6%) samples, respectively. The data signifies that Treponema sp. are significantly associated with these foot pathogens and their different combinations with Treponema sp. influence the severity of CODD lesion. The identification of Treponema phylotypes was done by sequencing the 16S rRNA gene fragment of ten representative samples. Out of ten sequences, four (Trep-2, Trep-4, Trep-7 and Trep-10) were identical to Treponema sp. phylotype 1 (PT1) that belongs to phylogroup T. refringens-like, one sequence (Trep-1) was genetically close (90% sequence homology) to Treponema brennaborense while five sequences (Trep-3, Trep-5, Trep-6, Trep-8 and Trep-9) matched with uncultured bacterium clones of treponemes forming separate monophyletic group in phylogenetic tree and could represent new digital dermatitis phylogroup presently containing five ovine specific phylotypes. This is the first report on the presence of Treponema phylotypes other than three digital dermatitis (DD) Treponema phylogroups viz. T. phagedenis-like, T. medium/T. vincentii-like, and T. pedis-like that are frequently detected in CODD lesions. Metagenomic analysis of two representative samples revealed the abundance of genus Treponema in CODD lesion while this genus was absent in swab collected from clinically healthy foot suggesting that it might play primary role in producing CODD. These findings may further aid in understanding the etiopathogenesis of CODD and could help to develop appropriate treatment and mitigation strategies to combat the disease.
Subject(s)
Cattle Diseases , Digital Dermatitis , Sheep Diseases , Sheep , Animals , Cattle , Digital Dermatitis/epidemiology , Digital Dermatitis/microbiology , Lameness, Animal , Phylogeny , RNA, Ribosomal, 16S/genetics , Treponema/genetics , Sheep, Domestic/genetics , Sheep Diseases/microbiology , Cattle Diseases/microbiologyABSTRACT
The paper describes the setting up and long-term continuous operation of the first real-life, pilot scale, sewage treatment plant based on the recently patented phytoremediation technology, trademarked as SHEFROL®. The unit was about three times cheaper to install, operate and maintain than the least expensive of the other wetland-based technologies presently in vogue. Its semi-permanent version is 30 times cheaper. Monitoring of flow rates and levels of treatment intermittently over a 3 year course of continuous operation indicated the constancy and robustness of the reactor in treating total solids, suspended solids, chemical oxygen demand, biological oxygen demand, total Kjeldahl nitrogen, and soluble phosphorous to the average extents of 94, 84, 79, 70, 62 and 28% respectively. Earlier experience with bench-scale SHEFROL® units has indicated that removal of metals like Cu, Ni, Co, Zn, and Mn also takes place to the extent of 25-45% in these systems. These primary, secondary, and tertiary treatments occurred in a single unit process with no necessity of any pumping, aeration, or recycling. Models based on artificial intelligence were developed which enable forecasting the reactor performance in terms of secondary and tertiary treatment, respectively.
The paper describes the setting-up, long-term (3 years) performance evaluation at pilot scale, and modeling of a recently patented novel and unprecedented phytoremediation-based sewage treatment technology. The system is seen to be much more robust, versatile, space efficient, and economical, than existing macrophyte-based sewage treatment systems.
Subject(s)
Sewage , Waste Disposal, Fluid , Biodegradation, Environmental , Artificial Intelligence , Bioreactors , NitrogenABSTRACT
The Himalayan ecosystem is critical for ecological security and environmental sustainability. However, continuous deforestation is posing a serious threat to Himalayan sustainability. Changing land-use systems exert a tenacious impact on soil carbon (C) dynamics and regulate C emissions from Himalayan ecosystem. Therefore, this study was conducted to determine the changes in different C pools and associated soil properties under diverse land-use systems, viz. natural forest, natural grassland, maize field converted from the forest, plantation, and paddy field of temperate Himalaya in the surface (0-20 cm) and subsurface (20-40 cm) soils. The highest total organic carbon (24.24 g kg-1) and Walkley-black carbon contents (18.23 g kg-1), total organic carbon (45.88 Mg ha-1), and Walkley-black carbon stocks (34.50 Mg ha-1) were recorded in natural forest in surface soil (0-20 cm depth), while soil under paddy field had least total organic carbon (36.45 Mg ha-1) and Walkley-black carbon stocks (27.40 Mg ha-1) in surface soil (0-20 cm depth). The conversion of natural forest into paddy land results in 47.36% C losses. Among the cultivated land-use system, minimum C losses (29.0%) from different pools over natural forest system were reported under maize-filed converted from forest system. Land conversion causes more C losses (21.0%) in surface soil (0-20 cm depth) as compared to subsurface soil. Furthermore, conversion of forest land into paddy fields increased soil pH by 5.9% and reduced total nitrogen contents and microbial population by 28.0% and 7.0%, respectively. However, the intensity of total nitrogen and microbial population reduction was the lowest under maize fields converted from the forest system. The study suggested that the conversion of natural forest to agricultural land must be discouraged in the temperate Himalayan region. However, to feed the growing population, converted forest land can be brought under conservation effective maize-based systems to reduce C loss from the intensive land use and contribute to soil quality improvements and climate change mitigation.
Subject(s)
Carbon , Ecosystem , Agriculture , Carbon/analysis , Forests , India , Nitrogen/analysis , Soil/chemistry , Zea maysABSTRACT
The aim of this study was to identify the immunodominant outer membrane proteins (OMPs) of Fusobacterium necrophorum from sheep affected with severe foot-rot. The OMP profile of ovine strains of F. necrophorum has not been well studied. We analyzed the OMP profile of the most frequent lktA variant JKS-F3 of F. necrophorum associated with severe ovine foot-rot with lesion score 4 in order to identify its major immunodominant OMPs. Electrophoretic separations of extracted OMPs showed a number of spots in two-dimensional electrophoretic gels. Two immunoreactive proteins of size around 43 kDa were identified through western blotting using hyperimmune sera raised in rabbits. These two immunogenic OMPs were analyzed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF/MS) which revealed that these two OMPs of lktA variant JKS-F3 of F. necrophorum showed 46 and 42 percent protein sequence coverage and scores of 125 and 114, respectively, with the reported 43 kDa outer membrane protein of F. necrophorum strain H05, a putative porin having properties similar to pore-forming proteins of anaerobic Gram-negative bacteria. These identified immunogenic OMPs will contribute to our understanding of the pathogenic role played by this organism in ovine foot-rot and could be exploited to devise an effective control strategy through development of an OMP-based recombinant vaccine to mitigate foot-rot in sheep and goats.
Subject(s)
Foot Rot , Fusobacterium necrophorum , Animals , Bacterial Outer Membrane Proteins/genetics , Goats , Membrane Proteins , Rabbits , Sheep , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Bovine genital campylobacteriosis caused by Campylobacter fetus subsp. venerealis (Cfv) is of considerable economic importance to the cattle industry worldwide. Cfv causes syndrome of temporary infertility in female cattle, early embryonic mortality, aberrant oestrus cycles, delayed conception, abortions and poor calving rates. In the present study, a total of 200 samples obtained from vaginal swabs, cervicovaginal mucous (CVM), preputial washes and semen straws were investigated that were obtained from organized cattle farm of MLRI, Manasbal and unorganized sectors. Out of a total of 200 samples, 49 (47·57%) vaginal swabs, 1 (3·33%) preputial wash and 8 (25%) carried out CVM samples were positive for Cfv, whereas none of the semen straws were positive for Cfv. A total of eleven isolates of Cfv were recovered. PFGE (Pulse field gel electrophoresis) analysis revealed four different pulsotypes (I-IV) circulating in the screened farms. A common pulsotype circulating among farms could not be established. Insertion element (ISCfe1), a 233 bp amplicon of Cfv, was sequenced and the sequence was deposited in GenBank (accession no: MK475662).
Subject(s)
Campylobacter Infections/veterinary , Campylobacter fetus/drug effects , Campylobacter/drug effects , Cattle Diseases/microbiology , Animals , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Campylobacter/classification , Campylobacter/genetics , Campylobacter/isolation & purification , Campylobacter Infections/microbiology , Campylobacter fetus/classification , Campylobacter fetus/genetics , Campylobacter fetus/isolation & purification , Cattle , DNA Transposable Elements , Drug Resistance, Bacterial , Farms , Female , Genotype , India , MaleABSTRACT
The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.
Subject(s)
Bacterial Vaccines/immunology , Dichelobacter nodosus/physiology , Dichelobacter nodosus/pathogenicity , Foot Rot/prevention & control , Gram-Negative Bacterial Infections/veterinary , Sheep Diseases/prevention & control , Vaccination/veterinary , Animals , Dichelobacter nodosus/genetics , Dichelobacter nodosus/immunology , Foot Rot/microbiology , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/prevention & control , India/epidemiology , Prevalence , Seroepidemiologic Studies , Serogroup , Sheep , Sheep Diseases/microbiology , VirulenceABSTRACT
The objective of this study was to determine the prevalence and identification of leukotoxin gene, lktA, variant strains of Fusobacterium necrophorum in the footrot lesions of sheep. The detection of F. necrophorum was carried out by PCR targeting the lktA gene fragment and identification of lktA variant strains was done by PCR-single stranded conformational polymorphism (PCR-SSCP) and gene sequencing. Of the 450 swabs collected from footrot lesions of sheep, 117 were lktA-positive for F. necrophorum. Of the 50 swabs collected from apparently asymptomatic sheep, only one was lktA-positive for F. necrophorum. The overall prevalence of F. necrophorum in footrot affected sheep in Kashmir valley was 26%, and ranged from 20 to 34.8%, respectively. PCR-SSCP of lktA gene fragment analysis revealed three lktA variants, designated as JKS-F1/F2/F3, while two samples (1.7%) showed multiple lktA variant strains of F. necrophorum in a single footrot-affected sheep hoof. This appears to be the first report on the presence of more than one lktA variant of F. necrophorum in a footrot lesion of sheep. The JKS-F3 lktA variant was the most frequent (75.4%), followed by JKS-F2 (14.4%) and JKS-F1 (8.4%), respectively. Among the three lktA variants identified, JKS-F3 was detected in 74 (86.0%) samples from severe footrot affected sheep with a lesion score of 4. The data suggest that JKS-F3 is the predominant lktA variant of F. necrophorum and is associated with severe footrot in sheep. Hence, JKS-F3 may be a significant variant contributing to the severity and duration of the disease in sheep.
Subject(s)
Carrier State/veterinary , Exotoxins/genetics , Fusobacterium Infections/veterinary , Fusobacterium necrophorum/genetics , Polymorphism, Single-Stranded Conformational , Sheep Diseases/microbiology , Animals , Carrier State/microbiology , Fusobacterium Infections/microbiology , Fusobacterium necrophorum/isolation & purification , India , Polymerase Chain Reaction , Prevalence , Sequence Analysis, DNA , SheepABSTRACT
Side population (SP) cells mediate chemoresistance in leukemia. However, chemical inhibition approach to target SP cells has been poorly studied. Herein, we report the discovery of isatin derivatives of nicotinic acid amide as potent side population cell inhibitors. The selected derivatives showed superior potency over the reference drug verapamil. Furthermore, the treatment increased chemosensitivity and inhibited the cell proliferation on three different leukemic cell lines, K562, THP-1 and U937, suggesting that both SP and the bulk of leukemic cells are affected. Moreover, treatment with the most potent compound Nic9 reduced the expression of ABCG2, demonstrating that side population inhibition effect of the target derivatives is at least via ABCG2 inhibition. Importantly, the target derivatives induced erythrocyte/dendritic differentiation to leukemic cells mainly through Musashi/Numb pathway modulation.
Subject(s)
Isatin/pharmacology , Leukemia/drug therapy , Niacinamide/pharmacology , Side-Population Cells/drug effects , ATP Binding Cassette Transporter, Subfamily G, Member 2 , ATP-Binding Cassette Transporters/antagonists & inhibitors , ATP-Binding Cassette Transporters/physiology , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Resistance, Neoplasm , Humans , Leukemia/pathology , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/physiology , Neoplastic Stem Cells/drug effects , Nerve Tissue Proteins/analysis , Receptors, Nerve Growth Factor/analysisABSTRACT
BACKGROUND: Human placental villous cytotrophoblasts exhibit relative externalization of negatively charged moieties to the outer leaflet of the plasma membrane during the time of syncytialization rendering their reactivity to positively charged cationic antimicrobial peptides (CAMPs) during the window of implantation and early placentation. Vaginal administration of a synthetic CAMP, Ala(8,13,18)-magainin II amide (AMA) inhibited blastocyst implantation and early placentation in monkeys. Furthermore, the administration of AMA resulted in significant inhibition of cell differentiation, enhancement in apoptosis and loss of viability in first trimester placental villous cytotrophoblasts in primary culture. The present study examines the effect of in vitro application of different doses (0, 1, 10, 100, 1000 ng/ml) of AMA on the secreted cytokine profiles of cytotrophoblasts obtained from placental villi samples (n = 13) collected during 8-9 weeks of gestation and grown on three-dimensional collagen matrix in vitro. METHODS: A panel of forty-eight (48) cytokines in conditioned medium was analysed using multiplex immunoassays technique. Further, the steady state transcript levels of four cytokines (CCL4, CCL5, IL1B, IL6), the concentrations of which were affected by AMA in the isolated cytotrophoblasts, as well as, two cytokines (IL1A and TNF) which were not affected by AMA were estimated. Input list of cytokines secreted by cytotrophoblasts and showing differential secretion in response to AMA were used in enrichment analysis for the generation of biological networks. RESULTS: Placental cytotrophoblasts secreted 27 cytokines, 13 of which are affected by AMA in vitro with significantly decreased secretion of CCLs-2, 3, 4, 5, CXCLs-1 and 8, FGF2 and MCSF and that of IL1B, IL6 and MIF, and increased secretion of IL16 and IL-2RA. Of the above cytokines showing differential secretion, only IL-2RA, IL16 and MIF showed significant correspondence in the steady state expression of their respective transcript levels. Post-hoc Enrichment analysis revealed Toll-like receptor (TLR) mediated pathways were the top-scored target pathways that were affected by AMA. CONCLUSIONS: Administration of a CAMP causes shift in the balance of immune-inflammatory responses involving downstream pathways of TLRs in cytotrophoblast function. Further verification of functions of placental trophoblasts on administration of CAMP with pregnancy outcome is necessary.
Subject(s)
Antimicrobial Cationic Peptides/pharmacology , Cytokines/metabolism , Down-Regulation/drug effects , Inflammation/metabolism , Trophoblasts/drug effects , Cells, Cultured , Female , Humans , Pregnancy , Trophoblasts/metabolismABSTRACT
The present study records the first case of non-specificity of typing primers developed by Dhungyel et al. A strain of Dichelobacter nodosus (JKS-20G) isolated from ovine footrot in Kashmir, India, showed specificity for serogroup C and G primers. The fimA sequence of the strain turned out to be closer to serogroup G than C. The nucleotide sequence showed maximum homology of 92% with that of serotype G1 strain 238 and 95% with partial sequence available for serotype G2 strain VCS 1004. However, the deduced amino acid sequence of the fimbrial subunit gene of JKS-20G differed from strain 238 by 16 amino acids and by four amino acids from that of partial sequence of strain VCS 1004. This variation indicates towards declaring this isolate as a new serotype (G3) but just insufficient to classify this into a new serogroup. Some of the amino acid substitutions were located within three hypervariable regions a characteristic of different serogroups. However, to ascertain whether this isolate deserves a new serotype status, there is a need to go for antigenic characterisation of this isolate using the tube and cross tube agglutination test.
Subject(s)
DNA Primers/genetics , Dichelobacter nodosus/classification , Foot Rot/microbiology , Gram-Negative Bacterial Infections/veterinary , Sheep Diseases/microbiology , Amino Acid Sequence , Animals , Bacterial Proteins/genetics , Base Sequence , Cloning, Molecular , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Dichelobacter nodosus/genetics , Dichelobacter nodosus/isolation & purification , Genetic Variation , Gram-Negative Bacterial Infections/microbiology , India , Molecular Sequence Data , Polymerase Chain Reaction/veterinary , Sensitivity and Specificity , Sequence Alignment , Sequence Analysis, DNA/veterinary , Serotyping/veterinary , SheepABSTRACT
The present study records the strain-specific molecular typing system for Dichelobacter nodosus (D. nodosus) based on genetic analysis of fimA locus. Based on the study two new serotypes B5 and B6 are reported within the serogroup B. Out of 200 swab samples collected randomly from foot lesions of footrot affected sheep from all the districts of Kashmir, India, 122 (61.0%) detected positive for D. nodosus. Serogroup B was predominantly prevalent in 83.60% of positive samples. Restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR) amplified fimA gene of D. nodosus serogroup B revealed only two fingerprint patterns (FP) designated as FP1 and FP2. The FP1 was most prevalent and depicted by 82.35% of the samples with serogroup B while, FP2 was depicted by rest (17.65%) of the samples. Though the FP1 fimA sequence had the homology of 95% to D. nodosus fimA of serotype B4 isolate VRS 54, but there were 14 nucleotide differences and four nucleotide insertions/deletions in the coding sequence between these two strains resulting in eight amino acid substitutions in the fimbrial subunit. Similarly the FP2 fimA showed the sequence homology of 97% with D. nodosus fimA of serotype B2 isolate 183, with 10 nucleotide differences and three nucleotide insertions/deletions between these two sequences. This resulted in six amino acid substitutions, plus an amino acid length variation in the subunit protein. Thus it was presumed that these FP1 and FP2 strains represented new serotypes (B5 and B6, correspondingly) within the B serogroup as the degree of amino acid sequence difference with their nearest homologous strains was much greater than that within a serotype (0-5 amino acid differences), but comparable to that between serotypes (8-15 amino acid differences). This presumption was confirmed by cross tube agglutination test.
Subject(s)
Dichelobacter nodosus/classification , Amino Acid Sequence , Cloning, Molecular , DNA, Bacterial , Dichelobacter nodosus/genetics , Dichelobacter nodosus/isolation & purification , Fimbriae Proteins/genetics , Fimbriae, Bacterial/chemistry , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA , SerotypingABSTRACT
The present study determines the prevalence, economic impact of virulent footrot in central Kashmir, India, along with isolation and molecular characterization of Dichelobacter nodosus (D. nodosus) where so far no such work has been carried out. Over all 12.54% prevalence of footrot was recorded in central Kashmir with highest (15.84%) in district Srinagar, and least (10.89%) in district Budgam, while it was 13.28% in district Ganderbal. Overall economic impact of footrot was estimated to the tune of Rs 15.82 million annually to the sheep farming in central Kashmir. Out of 370 samples collected from footrot lesions of naturally infected sheep, 200 (54.05%) detected D. nodosus positive by polymerase chain reaction (PCR). Out of these, 132 (66.00%) samples carried serogroup B of D. nodosus, five (2.50%) serogroup E, one (0.50%) serogroup I, while, 53 (26.50%) had mixed infection of serogroups B and E, four (2.00%) of serogroups B and I, two (1.00%) of serogroups B and G and the remaining three (1.50%) samples harboured the mixed infection of serogroups B, E and I. Serogroup G was detected for the first time in India. Over all serogroup B was most frequent (97.0%) followed by E (30.5%), while serogoups I (4.0%) and G (1.0%) were least prevalent. A total of 265 D.nodosus strains were isolated out of which 194 (73.20%) were typed as serogroup B, 61 (23.01%) as serogroup E, eight (3.01%) as serogroup I and remaining two (0.75%) belonged to serogroup G. Out of 265 D. nodosus isolates, 164 (61.88%) possessed intA (integrase) gene, thus were considered as virulent strains. Serogroup wise intA gene was found in 121(62.37%) isolates of serogroup B, 36 (59.01%) of E, two (100%) of G and five (62.50%) of I. Out of 20 randomly selected isolates subjected to gelatin gel test, 16 isolates with intA gene produced thermostable protease while four isolates without intA gene revealed the production of thermolabile protease. This indicated a good co-relation between presence of intA gene and gelatin gel test in determination of the D. nodosus virulence. Thus the present investigation suggests the incorporation of serogroups B and E, based on their predominant prevalence, in the formulation of an effective bivalent vaccine to combat footrot in central Kashmir.
Subject(s)
Dichelobacter nodosus/classification , Dichelobacter nodosus/genetics , Foot Rot/epidemiology , Sheep Diseases/epidemiology , Animals , DNA, Bacterial/genetics , Dichelobacter nodosus/isolation & purification , Foot Rot/economics , India/epidemiology , Polymerase Chain Reaction , Prevalence , Serotyping , Sheep Diseases/economics , Virulence Factors/geneticsABSTRACT
Histone deacetylase 2 (HDAC2), an isozyme of Class I HDACs has potent imputations in actuating neurodegenerative signaling. Currently, there are sizeable therapeutic disquiets with the use of synthetic histone deacetylase inhibitors in disease management. This strongly suggests the unfulfilled medical necessity of plant substitutes for therapeutic intervention. Sulforaphane-N-acetyl-cysteine (SFN-N-acetylcysteine or SFN-NAC), a sulforaphane metabolite has shown significantly worthier activity against HDACs under in vitro conditions. However, the atomistic studies of SFN-NAC against HDAC2 are currently lacking. Thus, the present study employed a hybrid strategy including extra-precision (XP) grid-based flexible molecular docking, molecular mechanics generalized born surface area (MM-GBSA), e-Pharmacophores method, and molecular dynamics simulation for exploring the binding strengh, mode of interaction, e-Pharmacophoric features, and stability of SFN-NAC towards HDAC2. Further, the globally acknowledged density functional theory (DFT) study was performed on SFN-NAC and entinostat individually in complex state with HDAC2. Apart from this, these inhibitors were tested against three distinct cancer cell models and one transformed cell line for cytotoxic activity. Moreover, double mutant of HDAC2 was generated and the binding orientation and interaction of SFN-NAC was scrutinized in this state. On the whole, this study unbosomed and explained the comparatively higher binding affinity of entinostat for HDAC2 and its wide spectrum cytotoxicity than SFN-NAC.
Subject(s)
Acetylcysteine/chemistry , Antineoplastic Agents/chemistry , Histone Deacetylase 2/antagonists & inhibitors , Isothiocyanates/chemistry , Sulfoxides/chemistry , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Benzamides/pharmacology , Binding Sites , Catalytic Domain , Cell Line, Tumor , Cell Survival/drug effects , Density Functional Theory , Drug Stability , Histone Deacetylase 2/genetics , Histone Deacetylase 2/metabolism , Histone Deacetylase Inhibitors/chemistry , Histone Deacetylase Inhibitors/metabolism , Histone Deacetylase Inhibitors/pharmacology , Humans , Hydrogen Bonding , Molecular Docking Simulation , Mutagenesis , Pyridines/pharmacology , ThermodynamicsABSTRACT
AIMS: To determine the genetic diversity of group A rotaviruses in bovine calves in Kashmir, India. METHODS AND RESULTS: Of 200 diarrhoeic faecal samples collected from calves, aged between 0 and 6 months and screened by polyacrylamide gel electrophoresis (PAGE), 31 were detected positive for group A rotaviruses. On G and P genotyping by reverse transcriptase-polymerase chain reaction (RT-PCR), G10P[11] turned out to be predominant (80·64%) combination followed by G8P[11] (7·7%). One (3·84%) sample carried mixed infection of G8+G10P[11]. Two (7·7%) samples belonged to P[11] genotype, but their G genotype specificity could not be established. This study revealed the ambiguity in RT-PCR typing method. All the samples that turned out to be G10 by Isegawa et al. (1993; Mol Cell Probes7, 277) primers could be amplified by G3 specific primers of Gouvea et al. (1990; J Clin Microbiol32, 1338). However, on homology study of their VP7 gene sequence, the strains turned out to be G10. CONCLUSIONS: Rotavirus is prevalent in diarrhoeic calves in Kashmir, India, and G10P[11] is the predominant genotype in circulation. There is evidence of mixed infection. Even though RT-PCR method is the quick way to type the strains, there is need to generate more sequence data to improve the specificity of typing primers. SIGNIFICANCE AND IMPACT OF THE STUDY: Rotavirus is a significant cause of diarrhoea in calves. RT-PCR typing method needs to be supported by the sequence data, and there is need to re-evaluate the primers used for typing.
Subject(s)
Cattle Diseases/virology , Diarrhea/veterinary , Feces/microbiology , Genetic Variation , Rotavirus Infections/veterinary , Rotavirus/genetics , Rotavirus/isolation & purification , Animals , Cattle , Diarrhea/virology , Genotype , India , Molecular Sequence Data , Reverse Transcriptase Polymerase Chain Reaction , Rotavirus/classification , Rotavirus Infections/virology , Viral Proteins/geneticsABSTRACT
A total of 480 samples, comprising 429 faecal samples from healthy adult birds and 51 tissue samples from dead birds, were collected from four government poultry farms in the Kashmir valley from September 2007 to April 2008. In all, 33 Salmonella isolates were obtained. Of these, 28 (84.85%) isolates were Salmonella Gallinarum, 3 (9.09%) were Salmonella Enteritidis and the remaining 2 (6.06%) were Salmonella Typhimurium. All the isolates harboured the invA, sefA, stn and spvC virulence-specific genes. However, the sopB gene was found in only 90.9% of the isolates. Pulsed-field gel electrophoresis analysis of representative isolates revealed that the majority were related but a few belonged to different clones. The majority of the isolates were resistant to cefpodoxime, nalidixic acid and sulphadiazine and sensitive to chloramphenicol, cefotaxime and tetracycline. Isolation of multidrug-resistant Salmonella, including the zoonotically important serovars, revealed a potential threat not only to poultry but also to human health in Kashmir.
Subject(s)
Anti-Infective Agents/pharmacology , Poultry Diseases/microbiology , Salmonella Infections, Animal/microbiology , Salmonella/drug effects , Animals , Bacterial Proteins/genetics , Electrophoresis, Gel, Pulsed-Field/veterinary , Feces/microbiology , India/epidemiology , Microbial Sensitivity Tests/veterinary , Molecular Epidemiology , Poultry , Poultry Diseases/epidemiology , Salmonella/classification , Salmonella/genetics , Salmonella/pathogenicity , Salmonella Infections, Animal/epidemiology , Salmonella enteritidis/drug effects , Salmonella enteritidis/genetics , Salmonella enteritidis/pathogenicity , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Salmonella typhimurium/pathogenicity , Serotyping/veterinary , Virulence Factors/genetics , Viscera/microbiologyABSTRACT
AIMS: To study the prevalence and characterize atypical enteropathogenic Escherichia coli (EPEC) and Shiga toxin producing E. coli (STEC) in avian species in India. METHODS AND RESULTS: Two hundred and twelve faecal samples collected from 62 chickens, 50 ducks and 100 pigeons were investigated for the presence of stx(1), stx(2), eae and ehxA virulence genes by multiplex PCR. In all, 42 E. coli isolates (25 chicken, 2 duck and 15 pigeon) possessed at least one virulence gene. Out of these, nine (4.24%) isolates were STEC and 33 (15.56%) were EPEC. All isolates from duck and chicken were EPEC while among 15 pigeon isolates nine (60%) were STEC and six (40%) were EPEC. Among the STEC isolates four each carried stx(1) or stx(2) and one possessed both stx(1) and stx(2). Subtype analysis of stx revealed the presence of stx(2f) in four STEC isolates. None of the STEC isolates carried stx(1c), stx(2c), stx(2d) or stx(2e). Isolates carrying stx(2f) demonstrated vero cell toxicity. One each belonged to serogroup O17 and O78, while one was rough and the other untypeable. All EPEC isolates were atypical as they lacked bfpA. This appears to be the first report of detection of stx(2f) from India. CONCLUSIONS: The study established the presence of stx(1) and stx(2f) containing E. coli in pigeons and atypical EPEC in poultry in India. Pigeons might serve as vectors for transmission of STEC to environment and humans. SIGNIFICANCE AND IMPACT OF THE STUDY: Taking into account the close contact between fanciers and pigeons, these findings warrant a more critical appraisal of these zoonotic pathogens in pigeons and humans.
Subject(s)
Birds/microbiology , Enteropathogenic Escherichia coli/isolation & purification , Shiga Toxins/metabolism , Shiga-Toxigenic Escherichia coli/isolation & purification , Animals , Bacterial Typing Techniques , Enteropathogenic Escherichia coli/classification , Enteropathogenic Escherichia coli/genetics , Enteropathogenic Escherichia coli/metabolism , Feces/microbiology , India , Phylogeny , Shiga Toxins/genetics , Shiga-Toxigenic Escherichia coli/classification , Shiga-Toxigenic Escherichia coli/genetics , Shiga-Toxigenic Escherichia coli/metabolism , Virulence Factors/genetics , Virulence Factors/metabolismABSTRACT
OBJECTIVE: The objective of the present study was to evaluate oxidative DNA damage in peripheral blood leukocytes (PBLs) of patients with coronary artery disease (CAD) and to explore the relationship of oxidised purine and pyrimidine with oxidative stress. METHODS: The study participants (n = 100) included 50 patients and unrelated 50 age-, sex- and population-subgroup (Jat Sikhs)-matched healthy controls. Oxidative DNA damage using the modified enzymatic comet in PBLs, and malondialdehyde (MDA) levels, total oxidant status (TOS) and total antioxidant status (TAS) in blood serum samples using spectrophotometric methods was determined. RESULTS: The basal DNA damage of percent tail DNA (T-DNA%) was increased as were tail moment (TM) and olive tail moment (OTM). Oxidative DNA damage in terms of oxidised purines and oxidised pyrimidines was also significantly (p < .001) elevated in patients. Rather the advanced stages of CAD, unstable angina and acute myocardial infarction had significantly more basal and oxidative DNA damage (p < .05) compared to stable angina. MDA levels (p < .01) and TOS (p < .001) were increased significantly in patients with significant (p < .001) decrease in TAS. There was positive correlation of oxidised purines (T-DNA% r = 0.399, p = .004; TM r = 0.623, p = .001; OTM r = 0.456, p= .001) and of total oxidative damage (TM r = 0.515, p = .001; OTM r = 0.463, p = .001) with disease severity, and, with TOS (r = 0.279, p = .050) and negative with TAS (r = -0.341, p = .015). Multiple linear regression analysis revealed TOS and disease severity as independent predictors of oxidative DNA damage. CONCLUSIONS: There was significant increase in oxidative DNA damage and oxidative stress in CAD patients compared to levels in healthy controls.
Subject(s)
Coronary Artery Disease/genetics , DNA Damage/genetics , Oxidative Stress , Purines/blood , Pyrimidines/blood , Biomarkers/blood , Case-Control Studies , Comet Assay , Coronary Artery Disease/blood , Female , Follow-Up Studies , Humans , Male , Middle AgedABSTRACT
Myelinated fibers are organized into distinct domains that are necessary for saltatory conduction. These domains include the nodes of Ranvier and the flanking paranodal regions where glial cells closely appose and form specialized septate-like junctions with axons. These junctions contain a Drosophila Neurexin IV-related protein, Caspr/Paranodin (NCP1). Mice that lack NCP1 exhibit tremor, ataxia, and significant motor paresis. In the absence of NCP1, normal paranodal junctions fail to form, and the organization of the paranodal loops is disrupted. Contactin is undetectable in the paranodes, and K(+) channels are displaced from the juxtaparanodal into the paranodal domains. Loss of NCP1 also results in a severe decrease in peripheral nerve conduction velocity. These results show a critical role for NCP1 in the delineation of specific axonal domains and the axon-glia interactions required for normal saltatory conduction.
Subject(s)
Axons/physiology , Cell Adhesion Molecules, Neuronal , Drosophila Proteins , Membrane Glycoproteins/physiology , Membrane Proteins/physiology , Nerve Fibers, Myelinated/physiology , Nerve Tissue Proteins/physiology , Neuroglia/physiology , Neuropeptides/physiology , Optic Nerve/physiology , Receptors, Cell Surface/physiology , Sciatic Nerve/physiology , Aging , Animals , Cloning, Molecular , Drosophila , Female , Genomic Library , Heterozygote , Homozygote , Humans , Male , Membrane Glycoproteins/deficiency , Membrane Glycoproteins/genetics , Membrane Proteins/genetics , Mice , Mice, Knockout , Nerve Fibers, Myelinated/ultrastructure , Nerve Tissue Proteins/genetics , Neuropeptides/deficiency , Neuropeptides/genetics , Potassium Channels/physiology , Receptors, Cell Surface/genetics , Restriction MappingABSTRACT
AIMS: The present study was aimed to evaluate the association of C2850T, G1846A and C100T polymorphisms of the CYP2D6 with coronary artery disease (CAD) in North-West Indian population. METHODS: In this case-control study, 200 patients with CAD and 200 age-, gender- and ethnicity-matched healthy controls were genotyped for C2850T, G1846A and C100T polymorphisms of CYP2D6 using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: Genotype and allele distributions of C2850T and G1846A polymorphisms of the CYP2D6 were significantly different between cases and controls (pâ¯=â¯0.038, pâ¯=â¯0.021; pâ¯=â¯0.048, pâ¯=â¯0.012, respectively) whereas the distribution of genotype and allele for C100T polymorphism did not differ significantly (pâ¯=â¯0.098, pâ¯=â¯0.117, respectively). The 2850T and1846A variants were significantly associated with the increased risk of developing CAD, as observed from the odds ratios for the 2850â¯T/T and 1846 G/A genotypes (OR: 2.44, 95% CI: 1.99-4.99, pâ¯=â¯0.015 and OR: 1.62, 95% CI: 1.02-2.56, pâ¯=â¯0.041, respectively). Moreover, the recessive model in C2850T and the dominant model in G1846A are the best fit inheritance models to predict the susceptible gene effects (OR: 2. 07, 95% CI: 1.05-4.08, pâ¯=â¯0.031 and OR: 1.70, 95% CI: 1.10-2.62, pâ¯=â¯0.016, respectively). On gender stratification, these associations were observed only in females in addition to the C/T genotype of C2850T (OR: 2.52, 95% CI: 1.42-4.38, pâ¯=â¯0.001) and C100T (OR: 3.18, 95% CI: 1.52-6.67, pâ¯=â¯0.002). Furthermore, it is also observed that the CAT (OR: 2. 61, 95% CI: 1.07-6.34, pâ¯=â¯0.035) and TAC (OR: 15. 22, 95% CI: 1.97-117.58, pâ¯=â¯0.009) are high-risk haplotypes for CAD in the total group, whereas, the TGC (OR: 1. 64, 95% CI: 1.02-2.62, pâ¯=â¯0.042) and CAT (OR: 4. 21, 95% CI: 1.12-15.59, pâ¯=â¯0.035) haplotypes provide gender-specific risk in females. CONCLUSIONS: Our results indicate significant association of C2850T, G1846A and C100T polymorphisms of CYP2D6 with CAD especially in females of North-West Indian population.