xperimental Evaluation of the Possibility of using the LAL test to Determine Bacterial Endotoxins in Vaccines for Rabies Prophylaxis

Relevance. The presence of pyrogenic impurities in finished dosage forms of concentrated cultural anti-rabies vaccines produced in the Russian Federation is determined using pyrogenicity tests on rabbits (in vivo). In accordance with the decision of the Board of the Eurasian Economic Commission dated September 7, 2018 N 151 «On the approval of guidelines for drawing up a regulatory document on the quality of a medicinal product», one of the requirements for parenteral drugs is the determination of bacterial endotoxins. This document indicates that the regulatory documentation should include a test and an admissibility criterion for bacterial endotoxins (BE) using the horseshoe crab amoebocyte lysate technique.Aims. Experimental evaluation of the possibility of using the LAL-test to determine bacterial endotoxins in national vaccines for the prevention of rabies.Materials and methods. The research of the drug «Cultural antirabies vaccine concentrated inactivated purified» of national production was carried out in accordance with the National Pharmacopoeia of the Russian Federation, General Pharmacopoeia Monograph OFS.1.2.4.0006.15 in three modifications: gel-clot test: methods A, B; turbidimetric kinetic test: method C; chromogenic kinetic test: method D.Results. Was investigated 6 series of the national vaccines for the prevention of rabies from two national manufacturers (using three pharmacopoeial methods). LAL reagent produced by two companies (Charles River Endosafe® and Lonza). In order to confirm the reproducibility of the method, the gel-clot test was carried out at different time intervals by one or two operators. During the research was determined the possibility of using photometric methods (method C and D).Conclusions. The research proved the possibility of determining bacterial endotoxins by methods: gel-clot test (method A), turbidimetric kinetic test (method C) and chromogenic kinetic test (method D). Method B is recommended for quantitative analysis of vaccine without instrumental methods. During the research all national vaccines for the prevention of rabies was free from bacterial endotoxins (no more 25 EU/ml).

Detection of Pyrogens in Hormonal Implants Using the LAL Test

The State Pharmacopoeia of the Russian Federation, 14th edition states that implants are a sterile dosage form, and have to be tested for pyrogens. However, it does not provide details on how the test should be performed for this dosage form.The aim of the study was to develop a LAL test procedure for detection of bacterial endotoxins (BE) in implants, using the example of a goserelin product.Materials and methods: BE extraction from the implant surface into an aqueous medium was performed with subsequent BE detection in the extract by turbidimetric kinetic test. The implant was then dissolved in dimethyl sulfoxide, and the obtained goserelin solution was tested for BEs using the gel-clot test.Results: the analysis of the Russian and foreign pharmacopoeial approaches to pyrogenic substance detection in hormonal implants helped to develop two sample preparation procedures for determination of BE content (in the extract and the implant solution). It was demonstrated that the BE content in the water extract did not exceed 0.01 EU/mL and was less than 0.07 EU per implant. The BE content in the implant solution was less than 8.3 EU per 1 mg of goserelin, which is almost eleven-fold lower than the theoretically-derived limit.Conclusions: the authors developed two test procedures for BE detection in hormonal implants using the LAL test, which could be included in manufacturers’ product files. The first procedure involves testing of the water extract from the implant surface and establishes the BE limit of no more than 20 EU/product. The second procedure involves complete dissolution of the implant in dimethyl sulfoxide and establishes the limit of not more than 97.22 EU per 1 mg of goserelin.

Investigation of Vestibular Function in Adult Patients with Gitelman Syndrome: Results of an Observational Study

Gitelman syndrome (GS) is a rare salt-losing tubulopathy caused by an inactivating mutation in the SLC12A3 gene, encoding the thiazide-sensitive sodium chloride cotransporter (NCC). Patients with GS frequently complain of vertigo, usually attributed to hypovolemia. Because NCC is also located in the endolymphatic sac, we hypothesized that patients with GS might have vestibular dysfunction. Between April 2013 and September 2016, 20 (22%) out of 90 patients followed at the reference center complained of vertigo in the absence of orthostatic hypotension. Sixteen of them were referred to an otology department for investigation of vestibular function. The vertigo was of short duration and triggered in half of them by head rotation. Seven patients (44%) had a vestibular syndrome. Vestibular syndrome was defined: (1) clinically, as nystagmus triggered by the head shaking test (n = 5); and/or (2) paraclinically, as an abnormal video head impulse test (n = 0), abnormal kinetic test (n = 4) and/or abnormal bithermal caloric test (n = 3). Five patients had associated auditory signs (tinnitus, aural fullness or hearing loss). In conclusion, we found a high frequency of vestibular disorder in GS patients suffering from vertigo, suggesting a role of NCC in the inner ear. Referent physicians of these patients should be aware of this extrarenal manifestation that requires specific investigations and treatment.

Analyzing transient respirometric data by analytical algorithm for Monod kinetic parameters

This study aims to develop an analytical algorithm with oxygen update (Ou) data obtained from transient respirometric measurement. Based on Monod kinetics, this study formulates a novel two-phase analytical model for an oxygen uptake rate plot (OUR vs. Ou) obtained by respirometric techniques. The first phase is a hyperbolic equation relating to exogenous and endogenous respiration, while the second phase is a linear equation for endogenous respiration only. An algorithm was therefore developed to analyze four Monod parameters by locating the best phase-separating point at which the absolute average relative error (ARE) of OUR is minimized. An analysis using test data on acetate verified that the algorithm is capable of transient kinetic parameter estimation with an ARE below 5–10%. A sensitivity analysis on domestic wastewater coupled with a Monte Carlo simulation concluded that the kinetic test must be conducted at a relatively high initial substrate level (So/Xo ≧ 1 and So/Ks ≧ 10) for reliable parameter estimation. Moreover, it is crucial to conduct the kinetic test with sufficient and acclimated seed culture for the degradation of substrate. The results of this study can be used to develop an automatic transient kinetic analyzer with modern programmable respirometers.

Prediksi Laju Pembentukan Air Asam Tambang dengan Metode Column Leaching Test

Prediction of Acid Mine Drainage Forming using Method of Column Leaching Test. One of the environmental problems in coal mining activities is the formation of acid mine drainage. Prediction of the formation of acid mine drainage is important as an effort to control environmental impacts. Acid mine water occurs with interactions between potentially acid-forming material with oxygen, bacteria and water. Objective of study is to analyze the potential for acid mine drainage forming based on material characteristics. The research method was carried out using static and kinetic tests. The static test method classifies materials according to the ability to produce clean acids with observed parameters such as paste pH, total sulfur, Acid Neutralizing Capacity (ANC), Net Acid Generation (NAG), Maximum Potential Acid (MPA), and Net Acid Producing Potential (NAPP). The Kinetic test method predicts the rate of acid-forming of a material. The kinetic test uses the Column Leaching Test Method by using material with composition designed to represent field condition. The kinetic method parameters are pH, electrical conductivity, acidity, alkalinity, sulfate content, and dissolved metal content (Fe, Mn, and Cd). Results of the static test classified the material into NAF Non-Acid Forming (NAF), Potential Acid Forming (PAF) and Uncertain (UC) material categories. The results of the Column Leaching Method classified the material into categories of potential and no potential to form acid mine water. The columns that have the potential to form acid mine drainage occur in columns with large amounts of tonnage of PAF material or those in the upper layer so that it reacts with oxygen. The columns that have no potential to produce acid mine drainage in columns with PAF material are in the middle layer or mixed with NAF material.

CAFFEINE REMOVAL BY ACTIVATED CARBON FROM COFFEE GROUNDS WITH ALTERNATIVE ACTIVATION IN ALUMINUM CAPSULES

Activated carbon is used like adsorbent in removing gaseous and liquid pollutants besides purification and chemical recovery. However, its production has a high cost and has sought alternative precursors materials such as coffee grounds. The objective of this study was to determine the removal of caffeine in aqueous solution using activated carbon produced from coffee grounds by adsorption. The coffee grounds was impregnated with ZnCl2 and activated in aluminum capsules where it was used alcohol cotton and put fire to consume the oxygen inside. The adsorption test was performed with concentration of caffeine of 25 mg/L, 0.5 g of activated carbon in 100 mL samples, leaving the suspension under stirring for 120 minutes, in acid, neutral and basic pH. The pH range which obtained better removal was the neutral, removing 99.04% caffeine in 40 minutes. The kinetic test was performed in 20 minutes with the intraparticle model being the best fit.

Optimization of Preparation Conditions of Novel Adsorbent from Sugar Scum Using Response Surface Methodology for Removal of Methylene Blue

A novel and inexpensive adsorbent was prepared from sugar scum for the removal of methylene blue as an organic pollutant from aqueous solutions. The response surface methodology was used to study the effects of the calcination temperature and time on the yield and the methylene blue adsorption. In order to determine the optimal conditions of the preparation, the Doehlert design and desirability function were applied. The increase in calcination temperature increases the methylene blue adsorption and induces a reduction in yield. The optimal conditions have been identified to be a calcination temperature of 986°C and calcination time of 61 min. The characteristics of the obtained adsorbent were determined using SEM/EDX, and surface functions were obtained based on FTIR and pHpzc. The produced adsorbent had a porous structure and a pHpzc of 12.5. The results showed that the yield was 49.74% and the adsorption of methylene blue was 24.52 mg·g−1 with a contact time of 10 h determined by kinetic test. The sugar scum was found to be an effective material for the preparation of appropriate adsorbent for dye removal from wastewater.