Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3

Search results for: glucuronide

3 Effects of Post-sampling Conditions on Ethanol and Ethyl Glucuronide Formation in the Urine of Diabetes Patients

Authors: Hussam Ashwi, Magbool Oraiby, Ali Muyidi, Hamad Al-Oufi, Mohammed Al-Oufi, Adel Al-Juhani, Salman Al-Zemaa, Saeed Al-Shahrani, Amal Abuallah, Wedad Sherwani, Mohammed Alattas, Ibraheem Attafi

Abstract:

Ethanol must be accurately identified and quantified to establish their use and contribution in criminal cases and forensic medicine. In some situations, it may be necessary to reanalyze an old specimen; therefore, it is essential to comprehend the effect of storage conditions and how long the result of a reanalyzed specimen can be reliable and reproducible. Additionally, ethanol can be produced via multiple in vivo and in vitro processes, particularly in diabetic patients, and the results can be affected by storage conditions and time. In order to distinguish between in vivo and in vitro alcohol generation in diabetes patient urine samples, various factors should be considered. This study identifies and quantifies ethanol and EtG in diabetic patients' urine samples stored in two different settings over time. Ethanol levels were determined using gas chromatography-headspace (GC-HS), and ethyl glucuronide (EtG) levels were determined using the immunoassay (RANDOX) technique. Ten urine specimens were collected and placed in a standard container. Each specimen was separated into two containers. The specimens were divided into two groups: those kept at room temperature (25 °C) and those kept cold (2-8 °C). Ethanol and EtG levels were determined serially over a two-week period. Initial results showed that none of the specimens tested positive for ethanol or EtG. At room temperature (15-25 °C), 7 and 14 days after the sample was taken, the average concentration of ethanol increased from 1.7 mg/dL to 2 mg/dL, and the average concentration of EtG increased from 108 ng/mL to 186 ng/mL. At 2–8 °C, the average ethanol concentration was 0.4 and 0.5 mg/dL, and the average EtG concentration was 138 and 124 ng/mL seven and fourteen days after the sample was collected, respectively. When ethanol and EtG levels were determined 14 days post collection, they were considerably lower than when stored at room temperature. A considerable increase in EtG concentrations (14-day range 0–186 ng/mL) is produced during room-temperature storage, although negative initial results for all specimens. Because EtG might be produced after a sampling collection, it is not a reliable indicator of recent alcohol consumption. Given the possibility of misleading EtG results due to in vitro EtG production in the urine of diabetic patients.

Keywords: ethyl glucuronide, ethanol, forensic toxicology, diabetic

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2 Production of Camel Nanobodies against of Anti-Morphine-3-Glucuronide for the Development of a Biosensor for Detecting Illicit Drug

Authors: Shirin Jalili, Sadegh Hasannia, Hadi Shirzad, Afshin Khara

Abstract:

Morphine is one of the most medicinally important analgesics and narcotics. Structurally, it is classified as an alkaloid because of the presence of nitrogen. Its structure is similar to that of codeine, thebaine, and heroin. An immunoassay to accurately discriminate between these analogous alkaloids would be highly beneficial. A key factor for such an assay is specificity with high sensitivity, which is totally dependent on the antibody employed. However, most antibodies against haptens are polyclonal serum antibodies that exhibit significant cross-reactivities with closely related compounds. The camel-derived single-chain antibody fragments (VHH) are the smallest molecules with antigen-binding capacity, possessing unique properties compared to other conventional antibodies. In this study, a library containing the VHH genes of a camel immunized with with morphine conjugated BSA following phage display technology was generated. By screening the camel-derived variable region of the heavy chain cDNA phage display library with the ability to bind the desired hapten, we obtained some nanobodies that recognize this hapten. Phage display expression of the Nbs from this library and pannings against this hapten resulted in a clear enrichment of four distinct Nb-displaying phages with specificity for morphine that could be a potential target site for the development of new strategies for the development of a biosensor for detecting illicit drug.

Keywords: phage display, nanobody, Morphine-3, glucuronide, ELISA, biosensor

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1 Potential of Polyphenols from Tamarix Gallica towards Common Pathological Features of Diabetes and Alzheimer’s Diseases

Authors: Asma Ben Hmidene, Mizuho Hanaki, Kazuma Murakami, Kazuhiro Irie, Hiroko Isoda, Hideyuki Shigemori

Abstract:

Type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) are characterized as a peripheral metabolic disorder and a degenerative disease of the central nervous system, respectively. It is now widely recognized that T2DM and AD share many pathophysiological features including glucose metabolism, increased oxidative stress and amyloid aggregation. Amyloid beta (Aβ) is the components of the amyloid deposits in the AD brain and while the component of the amyloidogenic peptide deposit in the pancreatic islets of Langerhans is identified as human islet amyloid polypeptide (hIAPP). These two proteins are originated from the amyloid precursor protein and have a high sequence similarity. Although the amino acid sequences of amyloidogenic proteins are diverse, they all adopt a similar structure in aggregates called cross-beta-spine. Add at that, extensive studies in the past years have found that like Aβ1-42, IAPP forms early intermediate assemblies as spherical oligomers, implicating that these oligomers possess a common folding pattern or conformation. These similarities can be used in the search for effective pharmacotherapy for DM, since potent therapeutic agents such as antioxidants with a catechol moiety, proved to inhibit Aβ aggregation, may play a key role in the inhibit the aggregation of hIAPP treatment of patients with DM. Tamarix gallica is one of the halophyte species having a powerful antioxidant system. Although it was traditionally used for the treatment of various liver metabolic disorders, there is no report about the use of this plant for the treatment or prevention of T2DM and AD. Therefore, the aim of this work is to investigate their protective effect towards T2DM and AD by isolation and identification of α-glucosidase inhibitors, with antioxidant potential, that play an important role in the glucose metabolism in diabetic patient, as well as, the polymerization of hIAPP and Aβ aggregation inhibitors. Structure-activity relationship study was conducted for both assays. And as for α-glucosidase inhibitors, their mechanism of action and their synergistic potential when applied with a very low concentration of acarbose were also suggesting that they can be used not only as α-glucosidase inhibitors but also be combined with established α-glucosidase inhibitors to reduce their adverse effect. The antioxidant potential of the purified substances was evaluated by DPPH and SOD assays. Th-T assay using 42-mer amyloid β-protein (Aβ42) for AD and hIAPP which is a 37-residue peptide secreted by the pancreatic β –cells for T2DM and Transmission electronic microscopy (TEM) were conducted to evaluate the amyloid aggragation of the actives substances. For α-glucosidase, p-NPG and glucose oxidase assays were performed for determining the inhibition potential and structure-activity relationship study. The Enzyme kinetic protocol was used to study the mechanism of action. From this research, it was concluded that polyphenols playing a role in the glucose metabolism and oxidative stress can also inhibit the amyloid aggregation, and that substances with a catechol and glucuronide moieties inhibiting amyloid-β aggregation, might be used to inhibit the aggregation of hIAPP.

Keywords: α-glucosidase inhibitors, amyloid aggregation inhibition, mechanism of action, polyphenols, structure activity relationship, synergistic potential, tamarix gallica

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