Vitamin D Status of Very Low Birth Weight Neonates at Baseline and Follow-up after Daily Intake of 800 IU Vitamin D.

INTRODUCTION: Vitamin D deficiency (VDD) is rampant in neonates. Recommendations for supplementation are variable. METHODS: An observational study was done on less than 32 weeks of very low birth weight neonates to find prevalence of VDD (<20 ng/ml) at baseline; at 38 ± 2 weeks post-menstrual age (PMA) after daily intake of 800 IU vitamin D (vit D). Secondary objectives were to find determinants of VDD, to compare growth in deficient; vit D sufficient (VDS) neonates; to find vit D toxicity. RESULTS: Of 83 neonates, 81 (97.6%) were VDD at baseline and 5 (6%) at 38 ± 2 weeks PMA. Determinants for VDD at baseline were inadequate maternal sun exposure (p < 0.001) and vit D supplementation (p = 0.007). Factors for VDD at 38 ± 2 weeks PMA were male gender (p = 0.049), morbidities (p = 0.006), ventilation >24 h (<0.001), sepsis (p = 0.032), caffeine (p ≤ 0.001) and missed supplements (p < 0.001). Weight and length gain of VDD to VDS neonates were (6.70 ± 2.40 to 8.96 ± 2.21 g/day); (0.82 ± 0.34 to 1.08 ± 0.37 cm/week), respectively (p < 0.001). Head circumference gain (cm/week) of VDS; VDD neonates was 0.58 ± 0.09; 0.54 ± 0.06 (p = 0.054), respectively. No neonates developed vit D toxicity. CONCLUSIONS: In preterm VLBW neonates, the prevalence of VDD was 97.6% but decreased by >90% at 38 ± 2 weeks with a daily intake of 800 IU vit D. Inadequate maternal vit D intake and sun exposure determined low baseline vit D status of neonates. Male gender, morbidities, ventilation, sepsis, caffeine, missed vitamin D supplements were determinants of poor vit D status at follow-up. Weight gain and length increments were more in the VDS group.

Evaluation of antiviral activity of Ocimum sanctum and Acacia arabica leaves extracts against H9N2 virus using embryonated chicken egg model.

BACKGROUND: In the view of endemic avian influenza H9N2 infection in poultry, its zoonotic potential and emergence of antiviral resistance, two herbal plants, Ocimum sanctum and Acacia arabica, which are easily available throughout various geographical locations in India were taken up to study their antiviral activity against H9N2 virus. We evaluated antiviral efficacy of three different extracts each from leaves of O. sanctum (crude extract, terpenoid and polyphenol) and A. arabica (crude extract, flavonoid and polyphenol) against H9N2 virus using in ovo model. METHODS: The antiviral efficacy of different leaves extracts was systematically studied in three experimental protocols viz. virucidal (dose-dependent), therapeutic (time-dependent) and prophylactic (dose-dependent) activity employing in ovo model. The maximum non-toxic concentration of each herbal extracts of O. sanctum and A. arabica in the specific pathogen free embryonated chicken eggs was estimated and their antiviral efficacy was determined in terms of reduction in viral titres, measured by Haemagglutination (HA) and real time quantitative reverse transcription polymerase chain reaction (RT-qPCR) assays. RESULTS: All the extracts of O. sanctum (crude extract, terpenoid and polyphenol) and A. arabica (crude extract, flavonoid and polyphenol) showed significant virucidal activity, however, crude extract ocimum and terpenoid ocimum showed highly significant to significant (p < 0.001-0.01) decrease in virus genome copy numbers with lowest dose tested. Similarly, therapeutic effect was observed in all three extracts of O. sanctum in comparison to the virus control, nevertheless, crude extract ocimum and terpenoid ocimum maintained this effect for longer period of time (up to 72 h post-incubation). None of the leaves extracts of A. arabica had therapeutic effect at 24 and 48 h post-incubation, however, only the crude extract acacia and polyphenol acacia showed delayed therapeutic effect (72 h post-inoculation). Prophylactic potential was observed in polyphenol acacia with highly significant antiviral activity compared to virus control (p < 0.001). CONCLUSIONS: The crude extract and terpenoid isolated from the leaves of O. sanctum and polyphenol from A. arabica has shown promising antiviral properties against H9N2 virus. Future investigations are necessary to formulate combinations of these compounds for the broader antiviral activity against H9N2 viruses and evaluate them in chickens.

Care seeking for childhood pneumonia by rural and poor urban communities in Lucknow: A community-based cross-sectional study.

CONTEXT: Globally, around 16% of under-five children die due to pneumonia. Childhood pneumonia, if identified early is a readily treatable through low-cost antibiotics. Access to timely and appropriate care is a key action to control pneumonia. AIMS: The aim of the study was to understand the caregiver's care-seeking behavior for the management of childhood pneumonia among rural and urban poor communities in Lucknow. SETTINGS AND DESIGN: Rural areas and urban slums of Lucknow district, from September 2014 to August 2015. SUBJECTS AND METHODS: A community-based cross-sectional study. Total of 1065 under-five children were selected by multistage random sampling method. Caregivers of children (<5 years of age) were interviewed through pretested, semi-structured interview schedule. Information was gathered on episode of cough, difficult breathing, and or chest indrawing in child within 2 weeks preceding the survey, and caregiver's treatment seeking practices. RESULTS: Out of total 1065 children, 52 (%) had pneumonia within 2 weeks preceding the survey. At the onset of illness, difficult/fast breathing was identified by 86.5% caregivers, but majority of them did not perceive it as a serious condition and resort to home remedies. Only 9.6% sought appropriate care at onset of illness. Appearance of chest indrawing in the child was identified by caregivers as a serious condition and sought treatment from outside. The mean time taken from onset of illness to the seeking care from health facility was around 2½ days (2.39 ± 0.75). Qualified private practitioners (70.5%) were the preferred choice and majority (87.0%) of the children received antibiotic for pneumonia. However, even after perception of seriousness of the illness, 26.8% and 11.1% caregivers in urban slums and rural areas, respectively sought inappropriate care at the first consultation. CONCLUSIONS: Caregivers were unable to perceive the severity of fast breathing leading to the delayed initiation of the appropriate treatment. There is a need of community mobilization through behavior change communication interventions to promote early symptom recognition and appropriate care seeking for pneumonia.

De Novo Assembly of Bitter Gourd Transcriptomes: Gene Expression and Sequence Variations in Gynoecious and Monoecious Lines.

Bitter gourd (Momordica charantia L.) is a nutritious vegetable crop of Asian origin, used as a medicinal herb in Indian and Chinese traditional medicine. Molecular breeding in bitter gourd is in its infancy, due to limited molecular resources, particularly on functional markers for traits such as gynoecy. We performed de novo transcriptome sequencing of bitter gourd using Illumina next-generation sequencer, from root, flower buds, stem and leaf samples of gynoecious line (Gy323) and a monoecious line (DRAR1). A total of 65,540 transcripts for Gy323 and 61,490 for DRAR1 were obtained. Comparisons revealed SNP and SSR variations between these lines and, identification of gene classes. Based on available transcripts we identified 80 WRKY transcription factors, several reported in responses to biotic and abiotic stresses; 56 ARF genes which play a pivotal role in auxin-regulated gene expression and development. The data presented will be useful in both functions studies and breeding programs in bitter gourd.

Effect of supplementation of taurine or trehalose in extender on immunolocalization of tyrosine phosphoproteins in buffalo and cattle (Karan Fries) cryopreserved spermatozoa.

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post-thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris-citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm) or trehalose (100 mm), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT-154 (anti-phosphotyrosine antibody) and FITC-conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post-thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing-thawing process.

Purification and characterization of Mn-peroxidase from Musa paradisiaca (banana) stem juice.

Mn-peroxidase (MnP), a biotechnologically important enzyme was purified for the first time from a plant source Musa paradisiaca (banana) stem, which is an agro-waste easily available after harvest of banana fruits. MnP was earlier purified only from the fungal sources. The enzyme was purified from stem juice by ultrafiltration and anion-exchange column chromatography on diethylamino ethylcellulose with 8-fold purification and purification yield of 65%. The enzyme gave a single protein band in SDS-PAGE corresponding to molecular mass 43 kDa. The Native-PAGE of the enzyme also gave a single protein band, confirming the purity of the enzyme. The UV/VIS spectrum of the purified enzyme differed from the other heme peroxidases, as the Soret band was shifted towards lower wavelength and the enzyme had an intense absorption band around 250 nm. The K(m) values using MnSO4 and H2O2 as the substrates of the purified enzyme were 21.0 and 9.5 microM, respectively. The calculated k(cat) value of the purified enzyme using Mn(II) as the substrate in 50 mM lactate buffer (pH 4.5) at 25 degrees C was 6.7s(-1), giving a k(cat)/K(m) value of 0.32 microM(-1)s(-1). The k(cat) value for the MnP-catalyzed reaction was found to be dependent of the Mn(III) chelator molecules malonate, lactate and oxalate, indicating that the enzyme oxidized chelated Mn(II) to Mn(III). The pH and temperature optima of the enzyme were 4.5 and 25 degrees C, respectively. The enzyme in combination with H2O2 liberated bromine and iodine in presence of KBr and KI respectively. All these enzymatic characteristics were similar to those of fungal MnP. The enzyme has the potential as a green brominating and iodinating agent in combination with KBr/KI and H2O2.

Assessment of intracellular Ca2+, cAMP and 1,2-diacylglycerol in cryopreserved buffalo (Bubalus bubalis) spermatozoa on supplementation of taurine and trehalose in the extender.

In mammalian spermatozoa, intracellular calcium plays a major role in sperm functions like motility and capacitation. Cryopreservation-induced modifications to sperm membrane result in an influx of intracellular calcium affecting calcium-dependent intracellular signalling pathways. Intracellular calcium activates adenyl cyclase to produce cAMP that activates phospholipase A(2) (PLA(2) ) and phospholipase C (PLC) generating lysophosphatidyl choline, 1,2-diacylglycerol (DAG) and IP(3) , acting as intracellular secondary messengers required for sperm capacitation. Present study was designed to determine levels of intracellular calcium, cAMP and DAG in fresh and frozen-thawed buffalo spermatozoa cryopreserved in the presence and absence of taurine or trehalose. A total number of nine ejaculates from three randomly chosen buffalo bulls were cryopreserved in Tris-based egg yolk extender and thawed in warm water at 37°C. The cAMP was measured by enzyme immuno assay, and intracellular calcium was quantified using fluorescent dye FURA 2-AM. Total lipid was extracted from spermatozoa, and DAG was estimated using thin layer chromatography followed by spectrophotometric analysis. Intracellular calcium, cAMP and DAG levels in spermatozoa were significantly (p < 0.01) increased following cryopreservation as compared to fresh ejaculate. Addition of taurine or trehalose to the freezing medium significantly decreased (p < 0.01) the levels of intracellular calcium and cAMP in frozen-thawed spermatozoa. 1,2-diacylglycerol content was also decreased significantly (p < 0.01) in spermatozoa cryopreserved in presence of additives. Moreover, significant (p < 0.01) improvement in post-thaw motility, viability and membrane integrity of spermatozoa on addition of taurine or trehalose clearly indicated the reduced level of capacitation-like changes in buffalo spermatozoa.

In silico analysis of sequential, structural and functional diversity of wheat cystatins and its implication in plant defense.

Phytocystatins constitute a multigene family that regulates the activity of endogenous and/or exogenous cysteine proteinases. Cereal crops like wheat are continuously threatened by a multitude of pathogens, therefore cystatins offer to play a pivotal role in deciding the plant response. In order to study the need of having diverse specificities and activities of various cystatins, we conducted comparative analysis of six wheat cystatins (WCs) with twelve rice, seven barley, one sorghum and ten corn cystatin sequences employing different bioinformatics tools. The obtained results identified highly conserved signature sequences in all the cystatins considered. Several other motifs were also identified, based on which the sequences could be categorized into groups in congruence with the phylogenetic clustering. Homology modeling of WCs revealed 3D structural topology so well shared by other cystatins. Protein-protein interaction of WCs with papain supported the notion that functional diversity is a con-sequence of existing differences in amino acid residues in highly conserved as well as relatively less conserved motifs. Thus there is a significant conservation at the sequential and structural levels; however, concomitant variations maintain the functional diversity in this protein family, which constantly modulates itself to reciprocate the diversity while counteracting the cysteine proteinases.

Molluscicidal activity of Saraca asoca and Thuja orientalis against the fresh water snail Lymnaea acuminata.

The molluscicidal activity of bark powder of Saraca asoca, leaf powder of Thuja orientalis against the snail Lymnaea acuminata was studied. The molluscicidal activity of all the plant products was found to be both time and concentration dependent. The 96 h LC(50) of T. orientalis leaf powder against L. acuminata was 250.5mg/l. Ethanol extracts were more toxic than other organic extracts. The ethanol extract of T. orientalis leaf (24h LC(50): 32.74 mg/l) was more effective than that of S. asoca bark (24h LC(50): 82.38 mg/l). The 24h LC(50) of column purified fraction of T. orientalis leaf and S. asoca bark powder was 29.25 and 64.89 mg/l, respectively. Saponin and thujone were identified as active molluscicide components in the bark of S. asoca and leaf of T. orientalis, respectively. The product of S. asoca and T. orientalis may be used as potent molluscicides.

Biosorption studies of nickel on Parthenium hysterophorous ash.

The application of an aquatic weed, Parthenium hysterophorous, has been investigated for the removal of nickel from aqueous solutions. Parthenium hysterophorous, the weed was converted to ash and was used as an adsorbent for the removal of nickel(II) from aqueous solutions at different experimental conditions. The per cent removal of Ni increased from 67.30 to 97.54%, with the nickel(II) concentration decreasing from 477.21 to 67.83 mg L(-1) at 25 degrees C, pH 11.0. The removal was favoured at higher pH, with a maximum removal at pH 11.0. The effects of concentration and temperature are also reported. Batch adsorption kinetics are described by the Lagergren equation. The value of the rate constant of adsorption was found to be 6.82 x 10(-2) min(-1) at 67.83 mg L(-1) and 25 degrees C. The applicability of the Langmuir and Freundlich equations for the present system were tested at different temperatures, viz. 25, 50 and 75 degrees C, and the constants were calculated. Thermodynamic parameters indicate the exothermic nature of nickel(II) adsorption on P. hysterophorous ash. The adsorption capacity was found to be much better than other common adsorbents reported for the removal of nickel(II).

Kinetics of enzyme inhibition by active molluscicidal agents ferulic acid, umbelliferone, eugenol and limonene in the nervous tissue of snail Lymnaea acuminata.

Ferulic acid, umbelliferone (Ferula asafoetida), eugenol (Syzygium aromaticum) and limonene (Carum carvi) are active molluscicidal components that inhibited the activity of alkaline phosphatase and acetylcholinesterase in in vivo and in vitro exposure of Lymnaea acuminata. It was observed that ferulic acid, umbelliferone and eugenol are competitive and limonene is a competitive-non-competitive inhibitor of alkaline phosphatase. Ferulic acid and umbelliferone are competitive, whereas eugenol and limonene are competitive-non-competitive and uncompetitive inhibitors of acetylcholinesterase, respectively.

Enzyme inhibition by the molluscicidal agent Punica granatum Linn. bark and Canna indica Linn. root.

Sublethal in vivo 24 h exposure to (40% and 80% of 24 h LC(50)) active fractions of Punica granatum bark or Canna indica root or in combination with other plant-derived molluscicides significantly inhibited the activity of acetylcholinesterase, acid/alkaline phosphatase, Na(+)K(+)ATPase and lactic dehydrogenase in the nervous tissue of Lymnaea acuminata. The inhibition kinetics of these enzymes indicates that active fractions of both the plants caused a competitive inhibition of AChE, LDH, ALP, ACP and Na(+)K(+)ATPase.

N-palmitoylation of the radioprotective domain of interleukin-1 affords inhibition of LPS-induced nitric oxide generation.

Interleukin-1beta (IL-1beta), a cytokine involved in homeostatic processes such as the immune system and inflammatory reactions, is a potent inducer of nitric oxide. The nonapeptide of human IL-1beta (VQGEESNDK, position 163-171, specific radioprotective domain--SRD) has been shown to retain radioprotective, immunostimulatory, and adjuvant activities of the native molecule without any inflammatory and pyrogenic properties. Unlike the parent IL-1, SRD did not induce nitric oxide (NO) in control or irradiated RAW 264.7 cells nor did it affect inducible nitric oxide synthase (iNOS) as shown by ELISA based mRNA assay (Quantikine). A lipophillic derivative of the SRD (a palmitoyl residue at the amino terminus of the SRD) was synthesized (palmitoyl specific radioprotective domain, P-SRD) to find out if this structural derivatization would restore the NO-inducing ability of IL-1. Surprisingly, P-SRD not only did not induce NO, but significantly inhibited lipopolysaccharide (LPS) stimulated nitric oxide (NO) production. Quantikine studies indicated that P-SRD also inhibited iNOS in LPS stimulated macrophage cells, suggesting that decrease in NO production in the presence of P-SRD was the result of iNOS mRNA inhibition. These results indicate that N-palmitoylation of SRD may effectively ameliorate potentially fatal symptoms of LPS-induced endotoxemic hypotensive shock associated with IL-1 without inflammatory and pyrogenic toxic side effects.

Anticellular and immunosuppressive properties of ethanolic extract of Acorus calamus rhizome.

Modulation of immune response to alleviate disease has been of interest since long. Plant extracts have been widely investigated for possible immunomodulatory properties. We have evaluated the anticellular and immunomodulatory properties of ethanolic extract of Acorus calamus rhizome. This extract inhibited proliferation of mitogen (phytohaemagglutinin; PHA) and antigen (purified protein derivative; PPD)-stimulated human peripheral blood mononuclear cells (PBMCs). In addition, A. calamus extract inhibited growth of several cell lines of mouse and human origin. It also inhibited production of nitric oxide (NO), interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha). Intracytoplasmic interferon-gamma (IFN-gamma) and expression of cell surface markers, CD16 and HLA-DR, on human PBMC, were not affected on treatment with A. calamus extract but CD25 expression was down regulated. Our study demonstrates the antiproliferative and immunosuppressive potential of ethanolic extract of A. calamus rhizome in vitro.

Effect of single and binary combinations of plant-derived molluscicides on different enzyme activities in the nervous tissue of Achatina fulica.

Effect of single and binary treatments of plant-derived molluscicides on different enzymes--acetylcholinesterase (AChE), lactic dehydrogenase (LDH) and acid/alkaline phosphatase (ACP/ALP)--in the nervous tissue of the harmful terrestrial snail Achatina fulica were studied. Sublethal in vivo 24-h exposure to 40% and 80% LC(50) of Azadirachta indica oil, Cedrus deodara oil, Allium sativum bulb powder, Nerium indicum bark powder and binary combinations of A. sativum (AS) + C. deodara (CD) and CD + A. indica (AI) oils significantly altered the activity of these enzymes in the nervous tissue of Achatina fulica. The binary treatment of AS + CD was more effective against AChE, LDH, and ALP than the single ones. However, binary treatment of AI + CD was more effective against ALP.

Immunomodulation by ethanolic extract of Boerhaavia diffusa roots.

We have earlier reported that ethanolic extract of Boerhaavia diffusa, a plant used in Indian traditional system of medicine, significantly inhibits the cell proliferation. This led us to evaluate the immunomodulatory properties of this plant extract on various in vitro tests such as human natural killer (NK) cell cytotoxicity, production of nitric oxide (NO) in mouse macrophage cells, RAW 264.7, interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha), intracytoplasmic interferon-gamma (IFN-gamma) and expression of various cell surface markers on human peripheral blood mononuclear cells (PBMCs). Ethanolic extracts of B. diffusa roots inhibited human NK cell cytotoxicity in vitro, production of NO in mouse macrophage cells, IL-2 and TNF-alpha in human PBMCs. Intracytoplasmic IFN-gamma and cell surface markers such as CD16, CD25, and HLA-DR did not get affected on treatment with B. diffusa extract. Our study demonstrates immunosuppressive potential of ethanolic extract of B. diffusa.

Antilymphoproliferative activity of ethanolic extract of Boerhaavia diffusa roots.

Extracts of plants have been widely evaluated for possible antiproliferative and anticarcinogenic properties. The antiproliferative activity of ethanolic extract of Boerhaavia diffusa, a plant used in traditional medicine, was evaluated in several cells. It inhibited T cell mitogen phytohemagglutinin and concanavalin A-stimulated proliferation of human peripheral blood mononuclear cells (PBMC). It also inhibited purified protein derivative antigen-stimulated PBMC proliferation and human mixed lymphocyte culture. In addition, B. diffusa extract inhibited the growth of several cell lines of mouse and human origin, such as mouse macrophage cells (RAW 264.7), human macrophage cells (U937), human monocytic cells (THP-1), mouse fibroblast cells (L929), human embryonic kidney cells (HEK293), mouse liver cells (BNLCL.2), African green monkey kidney cells (COS-1), mouse lymphoma cells (EL-4), human erythroleukemic cells (K562), and human T cells (Jurkat). The present study has demonstrated the antiproliferative potential of ethanolic extract of B. diffusa in vitro.

Effect of active molluscicidal component of spices on different enzyme activities and biogenic amine levels in the nervous tissue of Lymnaea acuminata.

In vivo exposure of Lymnaea acuminata to thymol and [6]-gingerol (active molluscicidal components of Trachyspermum ammi and Zingiber officinale, respectively) indicates that they significantly alter acetylcholinesterase, lactic dehydrogenase, succinic dehydrogenase and cyto-oxidase activity in the nervous -tissue of snails. In vitro exposure showed that, except for acetylcholinesterase and lactic dehydrogenase, no significant changes were observed in cyto-oxidase and succinic dehydrogenase activity in the nervous tissue of L. acuminata. Sublethal exposure to thymol and [6]-gingerol reduced the levels of 5-hydroxytryptamine (5-HT) and dopamine (DA) in the nervous tissue of L. acuminata. There was, however, no significant change in the level of 5-hydroxy indol acetic acid (5-HIAA). Thymol and [6]-gingerol thus affects all the known neurotransmission mechanisms in the snail either separately or through a complex interaction between the different neurotransmitters. This may account for their toxicity to snails.

Experimental autoimmune uveitis as animal model for human posterior uveitis.

Uveitis is an intraocular inflammatory disease that mostly affects children and young adults. It is one of the major causes of blindness in young individuals in India and the world. It is responsible for about 10 per cent of total visual impairment. Unfortunately, etiological diagnosis is not evident in a majority of these patients. It is generally felt that autoimmune mechanism may be involved in so called 'idiopathic' cases which has led to search for the putative autoantigens in experimental animal models. It has been demonstrated that experimental autoimmune uveitis (EAU) can be elicited against several retinal proteins in rats, mice and sub-human primates. These include the S-antigen, a major protein on retinal photoreceptor cell, interphotoreceptor retinoid binding protein (IRBP) and several others. There are many similarities between clinical entities and the EAU, but the EAU differs from the clinical conditions in being self-limited, and requiring complete Freund's adjuvant for induction of the disease. The disease can be induced only in susceptible strains. Nevertheless, use of the EAU model has allowed for identification of disease causing epitopes of antigens and evaluation of disease modifying strategies which could be applied in clinical situations. There has been significant progress in this field, but still a lot more is required to be learnt to translate it into clinical practice.