Combinational IL-2/IL-15 induction does not further enhance IL-15-induced lymphokine-activated killer cell cytotoxicity against human leukemia/lymphoma cells

In vitro comparisons of induction of perforin (PFP), granzyme B (GRB), production of cytokines, and cell-mediated cytotoxicity by interleukin-2 (IL-2), interleukin-15 (IL-15), or combinational IL-2/IL-15-induced lymphokine-activated killer cells were studied in this study. Whereas IL-2-induction was associated with a decrease in cultured cell population over a 7-day period, IL-15 alone or in combination with IL-2 resulted in significant increase including cytotoxic T lymphocytes and subsets of CD56+ lymphocytes, particularly cytokine-induced killer and cytolytic natural killer-T lymphocytes. The overall PFP, GRB, and tumor necrosis factor-alpha expression in different subtypes were also significantly higher with IL-15 alone or in combination with IL-2 induction with resultant superior cytotoxicity compared to IL-2 treatment. There was no significant advantage of addition of IL-2 over IL-15 induction. These results offer further information on the cytotoxic potency of these cytokines and their mechanisms of action implicating potential use of IL-15 as part of cytokine adoptive immunotherapy.     

Multi-locus analysis of HLA class II genes in DR2-positive IDDM haplotypes in Finland

Abstract: In this study we characterized the haplotypes found in IDDM patients that normally confer resistance to the disease in order to localize the polymorphisms relevant for the protection. We studied 15 DR2-positive subjects with IDDM for their DRB1, DRB5 and DQB1 genes using RFLP, polymerase chain reaction (PCR), oligonucleotide typing, and in some specific cases direct sequencing after allele-specific PCR. In addition we analyzed 39 DR2-positive, IDDM non-associated haplotypes representing those haplotypes that are not inherited to probands and hence are present only in healthy family members. The frequency of the DRB1*1501-DRB5*0101-DQB1*0602 haplotype was slightly decreased among diabetic patients (80% vs. 92%). In addition, two unconventional haplotypes DRB1*1501-DRB5*0101-DQB1*05031 and DRB1*1501-DRB5*0101-DQB1*0502 were found in patients with IDDM while all the control ones were conventional. The sequencing of the DQB1*0602 allele present in IDDM haplotypes showed no differences when compared to the controls. These results support the primary but not absolute role of DQ in the protection against IDDM. An additional role of factors centromeric to DQB1 gene was suggested by findings based on the biallelic TaqI RFLP polymorphism of the DQA2 gene. All DR2-DQB1*0602 IDDM haplotypes were associated with the 2.1-kb fragment while in the control group the 2.1-kb and 1.9-kb fragments were evenly distributed.     

Protein A immobilization and HIgG adsorption onto porous/nonporous and swellable HEMA-incorporated polyEGDMA microspheres

Both non swellable and swellable poly(EGDMA/HEMA) microbeads were produced by suspension copolymerization. These microbeads were modified by immobilization of a spacer-arm (hexamethylene diamine (HMDA)) and protein A. The optimal values for modifications were as follows: sodium periodate concentration, 1.0 mgml(-1); HMDA concentration, 4 mgml(-1); and glutaraldehyde concentration, 0.070 microgml(-1). Adsorption of protein A onto the plain and periodate oxidized poly(EGDMA/HEMA) microbeads were very close to each other, and were 0.01-0.02 mg protein A on the 1-g Microbeads I and II, respectively. Protein A immobilization on poly(EGDMA/HEMA) microbeads were studied at different temperatures, times, and pHs using single protein solution containing different amounts of proteins. The optimal values for immobilization were as follows: the initial protein A concentration, 0.1 mgml(-1); temperature, 25 degrees C; pH, 9.5; and immobilization time, 120 min. Incorporation of protein A resulted in 1.420 and 1.825 mg protein A on the 1-g Microbeads I and II, respectively. HIgG adsorption capacity on the protein A-incorporated poly(EGDMA/HEMA) microbeads is 27 and 35 mg HIgGg(-1) polymer for Microbeads I and II, respectively.     

Relationship between Pollen Counts and Weather Variables in East-Mediterranean Coast of Turkey

BACKGROUND: Aeroallergen sampling provides information regarding the onset, duration and severity of the pollen season that clinicians use to guide allergen selection for skin testing and treatment. OBJECTIVES: This atmospheric survey reports (1) airborne pollen contributions in Adana in one-year period (2) pollen onset, duration and peak level (3) the relationship between airborne pollen and selected meteorological variables and; (4) effects on symptoms in pollen allergic children. METHODS: Pollen sampling was performed with a volumetric Burkard Spore Trap. Meteorological data were measured daily from April 2001 to April 2002. Asthma symptom scores were investigated in 186 pollen allergic children that were on follow up in pediatric allergy outpatient clinics during same period. RESULTS: Average measurements included 82.5% tree pollen, 7.7% grass pollen and 9.8% herb pollen 54 taxa were identified during one year. The most prominent tree pollens were Cupressaceae, Eucalyptus and Pinus. The most common herb was Chenopodiaceae pollen family. When airborne pollen levels were examined in relation to single meteorological conditions; daily variations in total pollen counts were not significantly correlated with any variable studied (humidity, rainfall, temperature and wind) (p > 0.05). On the other hand, statistically significant relationship between pollen concentration and symptom scores were found (p > 0.05). Positive correlations were seen between both Gramineae and Herb pollen, and humidity and rainfall from March to July. However, positive correlations were detected between tree pollen counts and temperature and humidity in May and June. CONCLUSION: This survey is the first volumetric airborne pollen analysis conducted in the survey area in Adana. This study suggested that the effects of weather on pollen count and symptom scores in this population could not be clearly identified with the evaluation of one-year data. However, pollen counts had effect on allergic symptoms in pollen allergic children. Examination of the complex interaction of multiple whether parameters would perhaps more fully elucidate the relationship between meteorology and aerobiology and provide the clinician with information necessary to forecast pollen prevalence. An awareness of the ever chancing, local aeroallergen patterns requires regular monitoring. Such awareness serves as a useful guide in the effective testing and treatment of atopic patients.     

Detection and genotyping of oocysts of Cryptosporidium parvum by real-time PCR and melting curve analysis

Several real-time PCR procedures for the detection and genotyping of oocysts of Cryptosporidium parvum were evaluated. A 40-cycle amplification of a 157-bp fragment from the C. parvum beta-tubulin gene detected individual oocysts which were introduced into the reaction mixture by micromanipulation. SYBR Green I melting curve analysis was used to confirm the specificity of the method when DNA extracted from fecal samples spiked with oocysts was analyzed. Because C. parvum isolates infecting humans comprise two distinct genotypes, designated type 1 and type 2, real-time PCR methods for discriminating C. parvum genotypes were developed. The first method used the same beta-tubulin amplification primers and two fluorescently labeled antisense oligonucleotide probes spanning a 49-bp polymorphic sequence diagnostic for C. parvum type 1 and type 2. The second genotyping method used SYBR Green I fluorescence and targeted a polymorphic coding region within the GP900/poly(T) gene. Both methods discriminated between type 1 and type 2 C. parvum on the basis of melting curve analysis. To our knowledge, this is the first report describing the application of melting curve analysis for genotyping of C. parvum oocysts.     

Cost-effective analysis of candidate genes using htSNPs: a staged approach

We have previously shown that the selection of haplotype tag single nucleotide polymorphisms (htSNPs) and their statistical analysis in a multi-locus transmission/disequilibrium test (TDT) results in a more cost-effective genotyping strategy in disease association studies of genes by minimising redundancy due to linkage disequilibrium between SNPs. Further savings can be achieved by the use of a two-stage genotyping strategy. This approach is illustrated here in conjunction with the multi-locus TDT in determining whether common alleles of the immune regulatory genes RANK and its ligand TRANCE (RANKL) are associated with type 1 diabetes (T1D). A saving of approximately 75% of potential genotyping reactions could be made with minimal loss of power. There was little evidence from our analysis for association between the TRANCE and RANK genes and T1D in the populations tested.     

The effects of vitamin A, pentoxyfylline and methylprednisolone on experimentally induced amyloid arthropathy in brown layer chicks.

The effects of vitamin A, pentoxyfylline and methylprednisolone on experimentally induced amyloid arthropathy were investigated. In this study, 175 1-day-old brown layer chicks were used. Throughout the study Group II (vitamin A) received high doses of vitamin A (75,000 IU/kg), whereas Group I (negative control), Group III (positive control), Group IV (pentoxyfylline) and Group V (methylprednisolone) received normal levels of vitamin A in the diet. At the fifth week, the experimental Groups II, III, IV and V were injected with Freund's adjuvant intra-articularly to induce amyloid arthropathy. Group IV received pentoxyfylline and Group V received methylprednisolone (10 mg/kg, intramuscularly) once. Joint and blood samples were examined 13 weeks after the injections. The values in Groups I, II, III, IV and V, respectively, were as follows: amyloid arthropathy formation (%), 0, 100, 87, 76, 66; serum amyloid A (ng/ml), 166+/-17, 607+/-40, 423+/-39, 342+/-27, 293+/-22; serum retinol (microg/dl): 59.75+/-3.8, 42.72+/-3, 59.24+/-3.6, 102+/-9.1, 101.3+/-12.3; heterophil/lymphocyte ratio: 0.504, 0.75, 0.75, 0.087, 0.44. In conclusion, it was observed that vitamin A enhanced the development of amyloid arthropathy and there were positive associations between amyloidosis, increased levels of serum amyloid A and increased numbers of tissue infiltrating macrophages. Methylprednisolone had a more successful inhibitory effect on amyloid arthropathy than pentoxyfylline.     

The effect of balloon angioplasty on hypertension in atherosclerotic renal-artery stenosis. Dutch Renal Artery Stenosis Intervention Cooperative Study Group

BACKGROUND: Patients with hypertension and renal-artery stenosis are often treated with percutaneous transluminal renal angioplasty. However, the long-term effects of this procedure on blood pressure are not well understood. METHODS: We randomly assigned 106 patients with hypertension who had atherosclerotic renal-artery stenosis (defined as a decrease in luminal diameter of 50 percent or more) and a serum creatinine concentration of 2.3 mg per deciliter (200 micromol per liter) or less to undergo percutaneous transluminal renal angioplasty or to receive drug therapy. To be included, patients also had to have a diastolic blood pressure of 95 mm Hg or higher despite treatment with two antihypertensive drugs or an increase of at least 0.2 mg per deciliter (20 micromol per liter) in the serum creatinine concentration during treatment with an angiotensin-converting-enzyme inhibitor. Blood pressure, doses of antihypertensive drugs, and renal function were assessed at 3 and 12 months, and patency of the renal artery was assessed at 12 months. RESULTS: At base line, the mean (+/-SD) systolic and diastolic blood pressures were 179+/-25 and 104+/-10 mm Hg, respectively, in the angioplasty group and 180+/-23 and 103+/-8 mm Hg, respectively, in the drug-therapy group. At three months, the blood pressures were similar in the two groups (169+/-28 and 99+/-12 mm Hg, respectively, in the 56 patients in the angioplasty group and 176+/-31 and 101+/-14 mm Hg, respectively, in the 50 patients in the drug-therapy group; P=0.25 for the comparison of systolic pressure and P=0.36 for the comparison of diastolic pressure between the two groups); at the time, patients in the angioplasty group were taking 2.1+/-1.3 defined daily doses of medication and those in the drug-therapy group were taking 3.2+/-1.5 daily doses (P<0.001). In the drug-therapy group, 22 patients underwent balloon angioplasty after three months because of persistent hypertension despite treatment with three or more drugs or because of a deterioration in renal function. According to intention-to-treat analysis, at 12 months, there were no significant differences between the angioplasty and drug-therapy groups in systolic and diastolic blood pressures, daily drug doses, or renal function. CONCLUSIONS: In the treatment of patients with hypertension and renal-artery stenosis, angioplasty has little advantage over antihypertensive-drug therapy.