Very fast analysis of impurities in immunoglobulin concentrates using conjoint liquid chromatography on short monolithic disks

Transferrin and albumin are often present in immunoglobulin G (IgG) concentrates and are considered as impurities. Therefore, it is important to determine their concentration in order to obtain a well-characterized biological product. Here, we describe their determination based on conjoint liquid chromatography (CLC). The established method combines two different chromatographic modes in one step: affinity and ion-exchange chromatography (IEC) combined in one column. Therefore, two CIM Protein G and one CIM quaternary amine (QA) monolithic disks were placed in series in one housing forming a CLC monolithic column. Binding conditions were optimized in a way that immunoglobulins were captured on the CIM Protein G disks, while transferrin and albumin were bound on the CIM QA disks. Subsequently, transferrin and albumin were eluted separately by a stepwise gradient with sodium chloride, whereas immunoglobulins were released from the Protein G ligands by applying low pH. A complete separation of all three proteins was achieved in less than 5 min. The method permits the quantification of albumin and transferrin in IgG concentrates and has been successfully validated.     

The Impact of Diet and Fibre Fractions on Plasma Adipocytokine Levels in Prediabetic Adults

The impact of diet and fibre fractions on adipocytokines in obese subjects with a risk of diabetes has not been investigated in detail yet. The purpose of the study is to evaluate the effects of a 12-month lifestyle intervention with different fibre profiles (resistant starch (RS)—rich fibre, or ordinary food fibre profiles) on adipocytokine levels. Fifty participants are divided into two groups (RS group and Fibre group). The groups differ only in the percentage of the recommended level of the RS consumed as a fraction of the same total fibre amount. The applied dietary intervention includes intake of 7531 KJ/daywith a total fibre portion of 25–35 g/dayfor both groups that includes 15 g/day of RS for the RS group only. The levels of leptin, adiponectin, apelin, resistin, tumor necrosis factor (TNF)-alpha and C-reactive protein (CRP) are measured, and their relationship to anthropometric and biochemical parameters is estimated. Along with significant body weight loss, only leptin is significantly reduced by 13% in the RS group while in the Fibre group, apelin levels are significant (−21%). Polynomial regression shows a negative correlation between RS intake and adiponectin (R2 = 0.145) and resistin level (R2 = 0.461) in the RS group. This study indicates the possibility that fibre fractions differently influence the outcome of lifestyle interventions, as well as their adipocytokine levels, in obese prediabetic adults.     

Alterations in serum cortisol, CD4+ CD8+ lymphocyte sub-population ration and T cell mediated suppression of immune responses in the malnutrition of anorexia nervosa

Serum cortisol and T lymphocyte sub-populations (CD3+, 4+ and 8+) were studied in 22 consecutively admitted patients with anorexia nervosa (AN) who had a mean weight loss of 30%. In addition Concanavalin A (Con A) mitogen induced T cell suppression of lymphocyte response to PPD (purified protein derivative of tuberculin antigen) was analysed. Increased serum cortisol concentrations were found in the AN-patients compared to the control group, with mean levels 654 and 418 nmol/l respectively. The relative numbers of CD4+ lymphocytes (mean 36.2%) and the CD4+ CD8+ ratio (mean 1.54) were significantly reduced (p < 0.05) in the AN-patients compared to the control group (mean 41.6% and 2.14 respectively). T cell mediated, Con A induced suppression of lymphocyte response to PPD was increased in AN-patients compared to the control group with low (1mug/ml) Con A concentration, but unchanged with high (5 mug/ml) Con A concentration. There was no correlation between serum cortisol concentrations and the numbers of T lymphocyte subpopulations or T cell suppressor activity. In contrast, a highly significant correlation existed between serum cortisol and the duration of AN (p < 0.002), but not with relative weight loss or anthropometric variables: triceps skin-fold (TSF) and arm muscle circumference (AMC). Immunological variables were not correlated with duration of disease. Thus, immunological alterations of the T cell system are detectable in AN, but are subtle and their clinical importance is not well known.     

Influence of Fermentation and Drying Materials on the Contamination of Cocoa Beans by Ochratoxin A

Ochratoxin A (OTA) is a mycotoxin produced mainly by species of Aspergillus and Penicillium. Contamination of food with OTA is a major consumer health hazard. In Côte d’Ivoire, preventing OTA contamination has been the subject of extensive study. The current study was conducted to evaluate the influence of fermentation and drying materials on the OTA content in cocoa. For each test, 7000 intact cocoa pods were collected, split open to remove the beans, fermented using 1 of 3 different materials, sun-dried on 1 of 3 different platform types and stored for 30 days. A total of 22 samples were collected at each stage of post-harvesting operations. The OTA content in the extracted samples was then quantified by high-performance liquid chromatography. OTA was detected in beans at all stages of post-harvesting operations at varying levels: pod-opening (0.025 ± 0.02 mg/kg), fermentation (0.275 ± 0.2 mg/kg), drying (0.569 ± 0.015 mg/kg), and storage (0.558 ± 0.04 mg/kg). No significant relationships between the detected OTA level and the materials used in the fermentation and drying of cocoa were observed.     

Stage determination and therapeutic decision in human African trypanosomiasis: value of polymerase chain reaction and immunoglobulin M quantification on the cerebrospinal fluid of sleeping sickness patients in Côte d'Ivoire

In human African trypanosomiasis (HAT), two disease stages are defined: the first, or haemo‐lymphatic stage, and the second, or meningo‐encephalitic stage. Stage determination forms the basis of therapeutic decision and is of prime importance, as the drug used to cure second‐stage patients has considerable side‐effects. However, the tests currently used for stage determination have low sensitivity or specificity. Two new tests for stage determination in the cerebrospinal fluid (CSF) were evaluated on 73 patients diagnosed with HAT in Côte d'Ivoire. The polymerase chain reaction (PCR) detecting trypanosome DNA (PCR/CSF) is an indirect test for trypanosome detection whereas the latex agglutination test detecting immunoglobulin M (LATEX/IgM) is an indicator for neuro‐inflammation. Both tests were compared with classically used tests, double centrifugation and white blood cell count of the CSF. PCR/CSF appeared to be the most sensitive test (96%), and may be of use to improve stage determination. However, its value for therapeutic decision appears limited, as patients whose CSF was positive with PCR were successfully treated with pentamidine. This result confirms those of previous works that showed that some patients with trypanosomes in the CSF could be treated successfully with pentamidine. LATEX/IgM, which depending on the cut‐off, showed lower sensitivity of 76% and 88%, but higher specificity of 83% and 71% for LATEX/IgM 16 and LATEX/IgM 8 respectively, appears more appropriate for therapeutic decision making.     

Comparison of quantitative light-induced fluorescence, digital photography and transverse microradiography for quantification of enamel remineralization

Background: Quantitative light‐induced fluorescence (QLF) and digital photography (DP) have been proposed as clinical methods for measuring changes in enamel mineral content. The aim of this study was to compare the ability of QLF and DP with the in vitro gold standard transverse microradiography (TMR) to measure the remineralization of enamel subsurface lesions. Methods: Subsurface lesions were formed in enamel (n = 40) and exposed to remineralization solutions for 10 days. Changes were analysed by DP, QLF and TMR to determine percentage changes in luminescence (%L), fluorescence (%F) and mineral content (%R), respectively and correlation between these parameters determined. Results: The correlations between TMR and QLF (r = 0.63), TMR and DP (r = 0.59), and DP and QLF (r = 0.64) were all moderate but statistically significant (p < 0.001). The variability in %L and, to a lesser extent, %F values significantly impacted on the potential role of DP and QLF as methods by which mineral content changes produced by remineralization treatments could be accurately measured. Conclusions: Both QLF and DP provided data that correlated moderately with TMR data. QLF images were easier to analyse, free of glare and had less variability compared with those produced using DP.     

Non-surgical approach to bleeding gastric ulcer

Bleeding gastric ulcers is a common reason for emergency upper endoscopy in Emergency Center of Clinical Center of Serbia. Randomized controlled trials have shown that endoscopic hemostasis is beneficial for patients with a bleeding peptic ulcer. Aim of this study was to analyze the frequency, etiological factors and localization of bleeding gastric ulcer. At the same time we were evaluated a degree of bleeding activity according to Forrest's classification and modality of performed endoscopic hemostasis. All patients who underwent upper gastrointestinal (UGI) endoscopy for bleeding gastric ulcer in Emergency Center (January 2001 - December 2005.) were identified from an endoscopy database and the clinical records were reviewed retrospectivel. A total of 3954 patients underwent UGI endoscopy for presumed acute UGI hemorrhage. More than thirty % of them (31.1)-1230 had an endoscopic diagnosis of bleeding gastric ulcer. We observed 1230 bleeding patients (60% male and 40% female) with a mean age of 64.3. The commonest localization of bleeding gastric ulcers was antrum (54 - 15%). Percentage of patients who received non-steroidal anti-inflammatory drugs (NSAIDs) and/or salicilates before bleeding was 54 6%. The main symptom was melaena, which was observed in 82, 44% of patients with bleeding gastric ulcer. According to Forrest's classification of bleeding activity, the most of patients had F IB and F III degree (23, 41% and 22, 76%). Injection endoscopic hemostasis was performed in 26.34% patients, which had active bleeding (F IA, F IB) Hemostasis was initially obtained in 96% of bleeding patients. Bleeding gastric ulcer is one of the commonest endoscopic diagnosis in Emergency Center of Clinical Center of Serbia. The most frequent etiology factor was no--steroid antinflammatory drugs and/or salicilates. Injection endoscopic hemostasis is a safe procedure with a low cost, and, if successful, substantially reduces the need for emergency surgery.