Interaction effects of sunflower oil and aflatoxin at graded levels in diet on performance, serum and tissue biochemical profile, organ weights and immuneresponse in broiler chicken.

The dietary supplementation of fat has great potential in countering the toxic effects of aflatoxin (AF) in chickens, but the issue was less researched upon. An experiment was conducted to evaluate the response of broiler chickens to graded levels of AF B1 (0, 150 and 300 ppb) and sunflower oil (SFO) (0, 1.5 and 3.0%) in the diet in a 3 × 3 factorial manner to understand their interaction effects. A total of 360 broiler chickens divided into 9 equal groups were fed the diets during 0 to 35 days of age, and their response was evaluated in terms of performance, serum biochemical profile, organ weights, liver fat content and bone mineralization. Sunflower oil at 1.5% in diet countered (P ≤ 0.01) the adverse effects of 150 ppb AF on body weight, whereas at 300 ppb AF, such a response was seen at the higher level (3%) of SFO. Aflatoxin decreased (P ≤ 0.01) feed intake by 4 and 11% at 150 and 300 ppb concentration, respectively at 35 days of age, which was increased (P ≤ 0.01) with each incremental level of SFO supplementation (by 3.0 and 8.8%, respectively at 1.5 and 3%, respectively). Serum protein concentration increased (P ≤ 0.01) by SFO supplementation only at the higher concentration (300 ppb) of AF (by 42.4%), whereas total cholesterol and triglyceride concentration, and immune response to SRBC inoculation increased (P ≤ 0.01) with SFO at either level of AF (by 16.8, 18.7 and 75.6% at 1.5% SFO and 33.1, 36.9 and 94.2% at 3.0% SFO, respectively at 35 days of age). Weights of the liver, giblets, kidneys and pancreas increased (P ≤ 0.01) by 23.2, 14.7, 34.2 and 16.9%, respectively, and thymus weight decreased (P ≤ 0.04) by 25.4% with 300 ppb AF, and SFO at 3% in diet countered the effect on weight of the liver and giblets. Fat deposition in the liver increased (P ≤ 0.01) as the concentration of AF increased in diet (by 9.4 and 17.3%, respectively at 150 and 300 ppb AF), which was significantly (P ≤ 0.05) countered by SFO at 3% in diet. Tibia bone Ca content increased by 2.4% (P ≤ 0.01) with SFO supplementation in AF-fed chickens. It is concluded that dietary SFO supplementation countered the adverse effects of AF in broiler chicks in a dose-dependent manner, and higher level of oil (3% in diet) was required at the higher concentration of AF (300 ppb) in diet.

Diminished IL-17A levels may protect filarial-infected individuals from development of rheumatoid arthritis and systemic lupus erythematosus.

Nematode infections have been observed to inversely correlate with autoimmune disorders. Recently, we have shown the absence of filarial infection in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) who live in filarial-endemic areas. The mechanism(s) by which filarial-infected individuals are protected against the development of RA or SLE are unknown. In mice CIA, an experimental model for RA, ES-62, an execratory product of rodent filarial nematode , has been shown to improve arthritis through suppression of the IL-17 pathway. A total of 160 individuals, 40 each of endemic normal, filarial-infected cases, SLE and RA patients, from filarial-endemic areas, were enrolled in the study. Plasma levels of IL17-A, IFN-α and TNF-α were quantified by enzyme-linked immunosorbent assay (ELISA). RA and SLE patients displayed significantly higher plasma IL-17A, IFN-α and TNF-α levels compared to endemic normal and infected individuals. Furthermore, IL-17A levels were significantly low in participants with filarial infection compared to endemic controls ( p < 0.05). Interestingly, plasma IL-17A levels correlated inversely with circulating filarial antigen (CFA) ( p = 0.004, Spearman r = -0.51). Filarial infection was associated with low plasma IL-17A levels, a mechanism by which it possibly protects individuals in filarial-endemic areas from the development of autoimmune disorders like RA and SLE.

Development of a recombinant hCG-specific single chain immunotoxin cytotoxic to hCG expressing cancer cells.

A large number of cancers express human chorionic gonadotropin (hCG) or its subunits ectopically. Patients harboring such cancers have poor prognosis and adverse survival. PiPP is a monoclonal antibody of high affinity and specificity for hCGß/hCG. Work was carried out to develop a PiPP based recombinant immunotoxin for the immunotherapy of hCG expressing cancers. Recombinant PiPP antibody was constructed in scFv format in which gene encoding the VH and VL domains were joined through a linker. This scFv gene was fused to the gene expressing Pseudomonas exotoxin (PE38), and cloned in a Escherichia coli based expression vector under the control of strong bacteriophage T7 promoter. Immunotoxin conjugating scFv(PiPP) and PE38, was expressed in E. coli as recombinant protein. Recombinant PiPP immunotoxin was purified from the bacterial cell lysate and tested for binding and killing of hCGß expressing lymphoma, T-lymphoblastic leukemia and lung carcinoma cells in vitro. Immunotoxin showed nearly 90% killing on the cells. This is the first ever report on recombinant immunotoxin for binding and cytotoxicity to hCG expressing cancer cells, and thus can be a potential candidate for the immunotherapy of hCG expressing cells.

Serum ferritin level correlates with SLEDAI scores and renal involvement in SLE.

INTRODUCTION: Ferritin is an acute-phase reactant that is elevated in various autoimmune disorders. Serum ferritin levels have been positively correlated with disease activity scores of rheumatoid arthritis and systemic lupus erythematosus (SLE). Further, enhanced levels of ferritin have also been reported in lupus nephritis. However, there are no reports from the Indian subcontinent. METHODS: Seventy-six female SLE patients, diagnosed on the basis of revised ACR criteria, and 50 healthy females, age matched from similar geographical areas, were enrolled in the present study. Serum levels of ferritin, IFN-α and IL-6 were quantified by enzyme-linked immunosorbent assay (ELISA). Clinical, biochemical, serological and other markers of disease activity (C3, C4 and anti-dsDNA) were measured by standard laboratory procedure. RESULTS: Serum ferritin levels were significantly higher in SLE patients compared to healthy controls (p < 0.0001). Ferritin levels positively correlated with SLE Disease Activity Index (SLEDAI) (p = 0.001, r = 0.35), anti-dsDNA (p = 0.001, r = 0.35), IFN-α (p < 0.0001, r = 0.51) and IL-6 (p < 0.0001, r = 0.65) and negatively correlated with C3 (p = 0.0006, r = -0.38) and C4 (p = 0.01, r = -0.28). Interestingly, serum levels of ferritin were positively associated with proteinuria (p = 0.001, r = 0.36), serum urea (p = 0.0004, r = 0.39) and serum creatinine (p = 0.0006, r = 0.38). CONCLUSION: Serum ferritin is an excellent marker of disease activity and renal dysfunction in SLE.

Effect of dietary inclusion of toasted guar (Cyamopsis tetragonoloba) meal as a source of protein on performance of White Leghorn layers.

Two experiments were conducted to study the effect of including toasted (120°C/35 min) guar meal (GM, Cyamopsis tetragonoloba) in the diet on performance and egg shell quality of White Leghorn (WL) layers. Totals of 2376 and 2816 layer chickens (Babcock, BV 300) were randomly distributed into 27 and 32 replicates with 88 birds each in Experiments 1 and 2, respectively. Three diets in Experiment 1 (0, 50 and 100 g GM) and 4 diets in Experiment 2 (0, 50, 100 and 150 g GM/kg) were prepared having similar concentrations of energy and protein. Each diet was fed ad libitum to 9 and 8 replicates, respectively, in Experiments 1 (from 53 to 68 weeks) and 2 (35 to 46 weeks of age). Compared to soya bean meal (SBM) GM contained similar concentrations of protein, but was deficient in all essential amino acids except arginine, which was 70% higher than in SBM. Total non-starch polysaccharide (NSP) content in GM (166 g/kg) was lower than that of SBM (179 g/kg). Amongst different NSP fractions, GM contained higher levels of arabans, xylans, mannans and glucans compared to SBM. The galactomannan gum content in GM was 46 g/kg. Egg production (EP), body weight (BW), food intake (FI), food efficiency (FE) and egg quality (shell weight, shell per cent, shell thickness, Haugh unit score, egg density and egg breaking strength) parameters were not affected by incorporating GM up to 100 g/kg diet in Experiment 1. However, egg weight (EW) and egg mass (EM) were reduced significantly in groups fed on 100 g/kg diet. In Experiment 2, EP and FE were not affected by incorporating GM up to 100 g/kg, but were reduced at 150 g/kg diet. FI, EW, BW and egg quality parameters were not affected by incorporating toasted GM up to 150 g/kg diet. Based on the results of both experiments, it is concluded that toasted GM can be included in WL layer diets up to 100 g/kg without affecting EP, FE, EW, EM, Haugh unit score, BW and egg shell quality parameters.

Effect of preservatives, temperature and storage duration on stability of nucleic acids of pleopod tissues of Penaeus vannamei (Boone) and screening of viral infections.

In shrimp farming, screening for economically significant viral pathogens in nucleic acids of shrimps is vital for disease surveillance programmes and further, to take necessary precautions to ensure the sustainability of the farms and thereby the shrimp industry. Different preservatives, temperature and storage durations of the pleopod tissues of Penaeus vannamei broodstock were tested to investigate its effect on the quality and quantity of the nucleic acids. The pleopods were subjected to two preservation regimes and the yield and stability of the extracted nucleic acids were monitored over a time period of 12 months. Stability of the nucleic acids was assessed with nested polymerase chain reaction, and the yield was checked spectrophotometrically. Data was analysed by performing two way ANOVA and Tukeys Paired test. Preservation treatments included storage at -20 degrees C and 5 degrees C in RNAlater and in 70% ethanol. Significant variation (P < 0.05) was observed in both DNA and RNA yield and stability from ethanol and RNAlater stored pleopods at 5 degrees C. However, the yield and stability did not differ (P > 0.05) in both the preservatives at -20 degrees C. The RNA was degraded and yielded lesser quantity when pleopod tissues were stored in ethanol at -20 degrees C than when stored in RNAlater during storage duration of 9 months. This study would help the shrimp farmers and researchers to adopt better preservation strategy, vital for shrimp disease surveillance programmes and for traceability studies in the event of any disease outbreak.

Effect of different concentrations of metabolisable energy and protein on performance of White Leghorn layers in a tropical climate.

1. An experiment was conducted to study the effects of feeding graded concentrations of metabolisable energy (ME) and crude protein (CP) on the performance of layers. Nine diets with three concentrations each of ME (10.04, 10.67 and 11.30 MJ/kg) and CP (150, 165 and 180 g/kg) in a 3 × 3 factorial arrangement of treatments were formulated. 2. A total of 5544 White Leghorn (WL) pullets (20 weeks of age) were housed in 4-bird colony cages and 22 adjacent cages constituted a replicate. Each diet was fed ad libitum to 7 replicates from 21 to 72 weeks of age. Production variables were recorded in 13 laying periods of 28 d each, and the data were pooled into three production phases, namely initial (21-32 weeks), peak (33-52 weeks) and post-peak (53-72 weeks). 3. No interaction was observed between ME and CP for egg production (EP), food intake (FI), food efficiency (FE), egg weight (EW), egg mass (EM) and body weight gain. 4. The EP, EW and EM during the initial phase of production were not affected by dietary ME concentrations, while the EW and EM improved with increasing concentrations of dietary CP from 150 to 165 g/kg. 5. During the peak production phase, improvements in EP (ME and CP), FI (ME), FE (ME, CP), EW (ME) and EM (ME, CP) were observed with increasing concentrations of energy and protein to 11.30 and 180 g/kg diet, respectively. 6. EP, EW and EM were unaffected by dietary variation in concentrations of ME and CP during post-peak production phase, but the FE improved and FI reduced with increasing dietary concentrations of these nutrients. 7. It is concluded that the optimum concentrations of ME for WL layers during the 21-32, 33-52 and 53-72 weeks of age are 11.30, 11.30 and 10.04 MJ/kg diet, respectively. The corresponding values for CP in diets are 180, 180 and 150 g/kg.

Evidence for the presence of white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild Penaeus monodon (Fabricius) broodstock, in the southeast coast of India.

A survey on the presence of the viruses of two economically significant diseases, white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild-collected Penaeus monodon broodstock, was conducted during different seasons of the year in two major coastal areas of southeast India. The broodstock were collected along the coast of Tamil Nadu and Andhra Pradesh during summer, premonsoon, monsoon and post-monsoon seasons for three consecutive years. A total of 7905 samples were collected and subjected to MBV screening, and 6709 samples that were screened as MBV negative were diagnosed for WSSV. MBV was detected using rapid malachite green staining and WSSV by nested polymerase chain reaction. Prevalence data of the viruses were analysed using the EpiCalc 2000 program at 95% confidence interval. Samples collected from the Andhra Pradesh coast displayed a slightly higher prevalence of WSSV and MBV infection than those collected from Tamil Nadu, although this difference was not statistically significant (P > 005). In addition, it was found that the prevalence of both WSSV and MBV infections fluctuated according to season. Data on prevalence of these viruses in broodstock would be useful to develop strategies for shrimp health management along the southeast coast of India.

Laboratory investigation of drinking water sources of Kangra, Himachal Pradesh.

A total of 70 drinking water sources including piped water supply (n = 36), ground water sources (n = 24, hand pumps and bore wells) and natural water sources (n = 10, springs/step-wells) from various parts of district Kangra, Himachal Pradesh were investigated for their suitability for drinking purpose by presumptive coliform test. Three samples were collected from each source during different parts of the year. Piped water sources (91.7%) were most contaminated followed by natural water sources (90%) and ground water sources (62.5%). 70.5% of the total water samples (n = 210) were positive for coliforms. All the three samples from 8.3% (n = 3), 37.5% (n = 9) and 10% (n = 1) piped water, ground water and natural sources respectively, were negative for coliform organisms. A variety of organisms including Proteus, Klebsiella, Citrobacter, Escherichia coli (E. coli), Pasteurella, Enterobacter and Serratia liquefaciens were isolated from water samples positive for coliforms in presumptive coliform test. Thermo-tolerant coliform organisms; Escherichia coli, Citrobacter, Klebsiella and Enterobacter were 71.2% (n = 52) of the total bacterial isolations. These findings suggest absence of adequate treatment and disinfection of the water sources supplying drinking water in district Kangra.

Prevalence of Escherichia coli and Salmonella spp. in ready-to-eat meat and meat products in Himachal Pradesh.

In the present study, a total of 85 ready to eat meat and meat products were collected from 11 tourist's places of Himachal Pradesh to examine the prevalence of Salmonella spp. and Escherichia coli during May 2010 till April 2011. Out of 33 samples, meat curry and 25 samples of non-veg momo, 4(12.12%) and 1(4.0%) were found to be positive for Escherichia coli. respectively. All the 5 E. coli isolates obtained were belonged to four different serotypes (O8, O89, O60 and O Rough). 40% of the E. coli isolates were found to be multi drug resistant (MDR) and maximum resistance was showed to Ampicilin. All the isolates were highly sensitive to Chloramphenicol. However, Salmonella spp. could not be isolated from any of the ready to eat meat and meat products. The present study reveals that the contamination of ready to eat foods of animal origin with E. coli could be an important factor of gastrointestinal illness in the consumers.

Ayurveda panchakarma treatment success in a case of chronic spontaneous urticaria non-responding to conventional medicine-A case study.

This is a case study of a 34-year-old male patient of Chronic Spontaneous Urticaria (CSU) with severe generalised itching and rashes all over the body since 17 years. The patient has no significant relief with conventional antihistamine therapies, AST (Autologous serum therapy) and Omalizumab (OMA). He was clinically assessed and managed with Ayurvedic Panchakarma procedures like Snehapana(therapeutic administration of unctuous formulation like ghee, oil), Vamana, Virechana and internal medication in line of Seethapitta Chikitsa. The patient was asked to monitor hives and itching daily for one week using a validated weekly Urticaria Activity Score (UAS7) at the baseline, after Vamana, after Virechana and after follow up. The Quality of life was assessed at the baseline and after follow-up using Chronic Urticaria Quality of Life Questionnaire (CU-Q2oL). The assessment showed complete remission in disease activity and improvement in Quality of life as per the scores. The hematological and biochemical investigations reflect the safety and efficacy of Ayurveda in management of CSU.

Characterization of two Indian native chicken breeds for production, egg and semen quality, and welfare traits.

The present study was conducted to characterize 2 important native chicken breeds from India and compare them on growth, production, egg and semen quality, and welfare traits. The Aseel breed showed (P < 0.001) higher BW at different ages; higher shank, radius, and toe lengths at 40 wk of age; and greater egg weights at 28, 32, and 40 wk of age than did the Kadaknath breed. The Kadaknath breed reached sexual maturity at an early age, and it had higher 40-wk egg production (P < 0.001). Higher egg specific gravity (P < 0.05) and higher albumen (P < 0.001) and shell (P < 0.009) percentages in the Kadaknath were observed, whereas the Aseel breed had a higher yolk index (P < 0.004), higher yolk percentage (P < 0.001), and higher yolk-to-albumen ratio (P < 0.001). Concentration of sperm (P < 0.01), live sperm counts (P < 0.05), and semen appearance scores (P < 0.05) were higher in the Aseel breed than in the Kadaknath breed. The Aseel breed showed a greater incidence of feather-pecking behavior under floor rearing, and this was negligible or mild in the Kadaknath breed. Broodiness under cage rearing was observed (8.42%) in the Aseel breed. With regard to welfare traits, male Aseel birds had a significantly shorter (P < 0.05) duration of tonic immobility (TI) than did male Kadaknath birds. Furthermore, male Aseel birds had a shorter (P < 0.001) duration of TI than did female Aseel birds, whereas female Aseel birds had a longer (P < 0.05) duration of TI than did female and male Kadaknath birds. From this study, it was concluded that the breeds differed on various growth, production, egg and semen quality, and behavioral traits, but not on welfare traits, although female Aseel birds exhibited a greater fear response.

Formulation of PEG-ylated L-asparaginase loaded poly (lactide-co-glycolide) nanoparticles: influence of Pegylation on enzyme loading, activity and in vitro release.

The present paper describes the advantage of PEG-ylation of L-asparaginase before encapsulation over its incorporation in the native form. During encapsulation a considerable amount of native protein undergoes denaturation and forms insoluble aggregates. In an effort to overcome this problem, L-asparaginase was PEG-ylated before subjecting it to the harsh conditions as encountered during double emulsion solvent evaporation technique. L-asparaginase was conjugated with succinimidyl succinate derivative of polyethylene glycol (SS-PEG, MW 5000) followed by characterization of the formed conjugate using size exclusion-HPLC and SDS PAGE. The PEG-ylated L-asparaginase consisted of different isomers from mono to multi PEG-ylated depending upon the number of Lysine residues (14 in case of L-asparaginase) with about 5% as native protein. The specific activity as retained after PEG-ylation was 62.84 +/- 8.2% and further about 82.7% of activity was recovered from the particles. Imitated studies with the native protein confirmed the enhanced stability of the conjugated protein when exposed to the organic solvent and sonication and showed comparatively less encapsulation efficiency due to increased hydrophilicity. Release profiles for native as well as conjugated proteins consisted of sustained release of about 66.66% and 44.45% in 28 days, respectively. The decrease in the release can be attributed to the increase in the molecular weight of the conjugated protein. The study finally proved that PEG-ylation protected the enzyme and prevented it from denaturation during encapsulation.

Effect of dietary α -tocopherol concentration on performance and some immune responses in broiler chickens fed on diets containing oils from different sources.

1. An investigation was carried out into the effects of dietary α-tocopherol (α-T) concentration and source of supplemental oil on performance, activity of anti-oxidative enzymes and some immune responses in broilers from day-old to 41 d of age. 2. Three dietary concentrations of α-T (10, 50 and 100 mg/kg) with three sources of supplemental oil (sunflower - SFO, palm - PMO and safflower - SAO) were provided using a 3 × 3 experimental design. 3. Body weight gain and food conversion efficiency were not affected by either interaction or concentrations of α-T and sources of oil in diet. 4. Concentrations of total protein, globulin, triglycerides and cholesterol in sera increased significantly with dietary α-T concentration irrespective of the source of oil. Significantly higher concentration of serum albumin was evident in broilers fed on the SFO-based diet and the concentration of globulin was higher in groups fed on those diets containing PMO and SAO. 5. The lipid peroxidation (LP), measured as MDA release, decreased with the concentration of α-T in a dose-related manner with SFO- and SAO-based diets, although not with the PMO-based diet. With different oil sources, LP was significantly lower with the PMO-based diet compared to the others. Activities of glutathione peroxidase and RBC catalase increased and heterophil: lymphocyte ratio was reduced with concentration of α-T for each source of oil tested. 6. Assays for humoral and cell-mediated immune responses indicated no effect of the source of dietary supplemental oil or interaction, although an increasing concentration of dietary α-T improved cell-mediated immune responses. 7. It is concluded that sunflower oil, palm oil and safflower oil can be used as sources of oil for broiler diets without having any effect on performance, immune responses or the activity of anti-oxidizing enzymes. Higher concentrations of dietary α-tocopherol (50 or 100 mg/kg) reduced lipid peroxidation activity and enhanced activities of anti-oxidative enzymes, they also improved the cell-mediated immune responses in commercial broilers.

Nutritional evaluation and utilisation of quality protein maize, Nityashree hybrid maize, and normal maize in broiler chickens.

Three experiments were conducted to determine the apparent metabolisable energy, apparent digestible amino acid values, and utilisation of three maize varities in chickens: quality protein maize (QPM), hybrid maize Nityashree (HMN) and normal maize (NM). There was no significant difference in the apparent metabolisable energy (AME) content amongst the three varieties of maize. Lysine and threonine digestibilities were significantly higher in QPM compared to either HMN or NM. No difference in the digestibilities of other amino acids occurred among the three different maize varieties. There was no difference between diets containing NM or HMN for body weight gain (21 and 40 d of age) and overall feed conversion ratio (0-40 d), but values were significantly higher for QPM (and Lys-supplemented NM) diets. The relative weights of dressed meat yield and giblets were unaffected by dietary replacement of NM with QPM, HMN, or Lys supplementation of the NM diet. However, abdominal fat content decreased and breast meat yield increased with both dietary replacement of NM with QPM and Lys supplementation of the NM diet. The concentration of protein in serum was significantly increased by dietary replacement of NM with QPM. Adding Lys to the NM based diet significantly increased the Ca concentration in serum compared with NM or HMN diets. However, the highest concentration of Ca in serum was found in the QPM diet. The concentration of total cholesterol in serum significantly decreased by either replacing NM with QPM or adding Lys to the NM based diet, compared with the NM diet. These results suggested that the feeding value of quality protein maize was superior to normal maize, while the feeding value of hybrid maize Nityashree was similar to that of normal maize.

Rice bran lysolecithin as a source of energy in broiler chicken diet.

1. Rice bran lysolecithin (RBL) was evaluated in broiler chicken diets. In the first experiment, RBL was included in diet at 0, 0·5, 2, 8 and 32 g/kg and fed to 250 broiler chickens from 0 to 42 d of age. In the second experiment, RBL was fed at 0, 25 and 50 g/kg diet to 405 day-old broiler chickens until 21 d of age, while during the finisher phase (22-35 d of age) chickens receiving each concentration of RBL were given all three concentrations of RBL in a 3 × 3 factorial manner. The diets were isocaloric. 2. Body weight, food consumption and food conversion efficiency were unaffected by feeding RBL, while the weight of pancreas increased at ≥2 g/kg of RBL in diet (experiment 1). In experiment 2, body weight was greater in the chickens receiving RBL at either 25 or 50 g/kg (21 d) and 50 g/kg (35 d of age). At 21 d of age, food consumption was greater at 25 or 50 g RBL/kg diet, while food conversion efficiency improved with 50 g RBL/kg diet. 3. Fat digestibility increased with RBL at 32 g/kg (experiment 1) and ≤25 g/kg (experiment 2). Rice bran lysolecithin increased ready to cook weight at 50 g/kg during starter phase and decreased abdominal fat at 25 and 50 g/kg during finisher phase (experiment 2). Liver and meat fat content were not affected. 4. It is concluded that lysolecithin from rice bran oil could be used as energy supplement in broiler chicken diet.

Lack of association between SNP rs3914132 of the RELN gene and otosclerosis in India.

Otosclerosis (MIM 166800) is primarily a metabolic bone disorder of the otic capsule, which leads to bony fixation of the stapedial footplate in the oval window; it is among the most common causes of acquired hearing loss. The etiology of this disease is largely unknown, although epidemiological studies suggest the involvement of both genetic and environmental factors. Recently, a reelin gene, SNP rs3914132, located in intron 2, was shown to be associated with otosclerosis in a European population. When we sequenced blood DNA samples of 85 individuals with otosclerosis and 85 controls, four SNPs of this gene: rs3914131 (P = 0.6463), rs3914132 (P = 0.1822), rs9641319 (P = 0.7371), and rs10227303 (P = 0.5669) were not significantly associated with this disease. In one familial case, a novel variant (C/T) at contig position 2923488 was found to be inherited by the proband and affected family members.