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.     

Sequence analysis of rpoB mutations in rifampin-resistant clinical Mycobacterium tuberculosis isolates from Turkey

Drug-resistant tuberculosis is a serious problem throughout the world. Resistance to Rifampicin (RIF) is mainly caused by the mutations in the rpoB gene coding the beta-subunit of RNA polymerase. In this study, we aimed to detect the distribution of rpoB gene mutations in 80 RIF-resistant clinical Mycobacterium tuberculosis (MTB) isolates from Turkey. The rpoB gene was amplified by PCR and mutations leading to RIF resistance were determined by automated sequence analysis. A total of 72 of the 80 isolates (90%) were found to carry mutations in the amplified region, whereas eight isolates (10%) carried no mutations. Overall, 24 different missense mutations affecting 14 codons, and two deletion mutants were identified. Nine new mutations, six in the hot-spot region and three outside this region, were found. The codon numbers of the most frequently encountered mutations were 531 (51.4%), 526 (18.1%), 516 (13.9%), and 513 (12.5%). As a result, 90% of the RIF-resistant MTB isolates from the Turkish patients were found to carry a mutation in the rpoB gene, Ser531Leu being the most frequent one. Although molecular methods identify mutations leading to RIF resistance very quickly, results of the antimycobacterial susceptibility tests must be taken into consideration for the patients carrying no mutations in this region.     

Neuropeptide Y in the female reproductive tract of the rat. Distribution of nerve fibres and motor effects

The spontaneous firing of single neurones in the region of the lateral reticular nucleus was the subject of a pharmacological study employing microiontophoretic and systemic application of adrenoceptor agonists and antagonists. Both iontophoretic noradrenaline and systemic clonidine depressed neuronal firing. The depressions were consistently reversed by the alpha-2 antagonist RX781094. Other adrenergic antagonists, prazosin and sotalol, were ineffective. The results suggest the existence of alpha-2 receptors in this region of the brain.     

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.     

The effect of Corynebacterium parvum in influenza infected mice

This paper is the continuation of earlier studies on the effect of the killed suspension of Corynebacterium parvum in influenza virus infected mice. Our investigation showed the normalized effect of these drugs on disturbed function of cell mediated immunity during experimental influenza infection especially in phagocytic and bactericidal activity of granulocytes. The present experiments concern the explanation of these infection mechanisms. Intraperitoneal injection of Corynebacterium parvum stimulated spleen index. Foot pad test is higher than in comparatively treated BCG group. The pathomorphological analysis of the spleen, thymus and peritoneal lymph nodes points out to the multiplication of multiple lymph nodes sinus cells. Generally, C. parvum possessed protective effect in experimental influenza infection. We tested the following parameters: phagocytic and bactericidal activity of granulocytes, liberation of leukocytes migration inhibition factor (LIF).     

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 diagnostic value of magnetic resonance imaging in subacromial impingement syndrome

The aim of this study was to assess the diagnostic ability of magnetic resonance imaging (MRI) in subacromial impingement syndrome (SIS), using a physiological standard of reference. MRI of the rotator cuff (RC) and subacromial injection test (SIT), a reference standard for SIS diagnosis, were performed in 125 painful shoulders. MRI diagnostic accuracies were determined using a 2 x 2 table and the percentage values of SIS diagnosis in patients with the three Zlatkin MRI stages were determined. Shoulder function was evaluated using the Constant Scale, and results were compared for stages. The sensitivity, specificity, accuracy, positive and negative predictive values of MRI for SIS diagnosis were 98.85%, 36.84%, 80%, 78.18% and 93.33% respectively. Of the 32 patients with Zlatkin stage 1 changes in MRI, 20 (62%) had SIT approved SIS diagnosis, while 47 (79%) of the 59 patients with Zlatkin 2 and all of the 19 (100%) patients with Zlatkin 3 changes were diagnosed with SIS by SIT. Mean Constant scores were 78.04 +/- 18.3, 65.0 +/- 19.9 and 54.52 +/- 20.7 in patients with Zlatkin stages 1, 2 and 3, respectively (p < 0.05). The MRI of RC did not prove to be an excellent tool for SIT based SIS diagnosis, with its low specificity. However, the technique can give important clues, as its sensitivity and negative predictive values are high.     

Relation of the radial nerve with the sulcus nervi radialis: a morphometric study

The radial nerve's course from the axillary region, branch patterns and the relation of the nerve to fixed anatomical landmarks in the arm region were studied in 27 embalmed intact cadavers. The radial nerve and its relation with the sulcus nervus radialis (SNR) was analyzed. The direct contact of the nerve with humerus in SNR was observed during the dissections. The following measurements were made: the total length of the humerus (the palpable uppermost point of the tuberculum majus and the lateral epicondyle); proximal safe zone (the tuberculum majus and the proximal beginning of the SNR); distal safe zone (the intercondylar axis and the middle of SNR); lateral safe zone (the lateral epicondyle and the distal end of SNR). In conclusion, it was aimed to correlate the osseus palpable landmarks of humerus with the course of the radial nerve for a safe surgery as the sulcus nervi radialis region is one of the main risky areas for the radial nerve palsies.     

Chromosome 22q11.2 deletion and phenotypic features in 30 patients with conotruncal heart defects

This report describes the dysmorphic features and frequency of 22q11.2 deletion (del22q11) in 30 Turkish patients with conotruncal heart defects (CTHDs). Fluorescence in situ hybridization (FISH) analysis revealed deletions in the 22q11.2 region in nine (30%) individuals. The CTHDs in this group were tetralogy of Fallot (four cases), double-outlet right ventricle (DORV) (two cases), transposition of great arteries (two cases), and ventricular septal defect (VSD) associated with other CTHDs (one case). The frequency of del22q11 in the study group was relatively high because many of the patients with dysmorphic findings also had cardiac anomalies involving the pulmonary artery, ductus arteriosus, or the aortic arch and its main branches. Twenty of the 30 patients exhibited several dysmorphic findings. Two of the nine patients with del22q11 exhibited no apparent dysmorphic features other than sacral dimple. Interestingly, one of the patients with del22q11 had a phenotypic appearance similar to that seen in oculo-auriculo-vertebral spectrum (OAVS). This individual had left microtia, atresia of the external meatus, mandibular asymmetry, and peripheral facial nerve paralysis. His mental development was normal and there were no abnormalities on ophthalmological examination. The CTHDs in this patient were situs inversus dextrocardia, DORV, pulmonary stenosis, and VSD. Radiographs of this patient showed platybasia, complete fusion of C2-C3, and posterior fusion of the T1-T2 vertebrae. This particular case indicates that the phenotypic features of del22q11 and OAVS may overlap.