Using BpyAla to generate copper artificial metalloenzymes: a catalytic and structural study.

Artificial metalloenzymes (ArMs) have emerged as a promising avenue in the field of biocatalysis, offering new reactivity. However, their design remains challenging due to the limited understanding of their protein dynamics and how the introduced cofactors alter the protein scaffold structure. Here we present the structures and catalytic activity of novel copper ArMs capable of (R)- or (S)-stereoselective control, utilizing a steroid carrier protein (SCP) scaffold. To incorporate 2,2'-bipyridine (Bpy) into SCP, two distinct strategies were employed: either Bpy was introduced as an unnatural amino acid (2,2'-bipyridin-5-yl)alanine (BpyAla) using amber stop codon expression or via bioconjugation of bromomethyl-Bpy to cysteine residues. The resulting ArMs proved to be effective at catalysing an enantioselective Friedel-Crafts reaction with SCP_Q111BpyAla achieving the best selectivity with an enantioselectivity of 72% ee (S). Interestingly, despite using the same protein scaffold, different attachment strategies for Bpy at the same residue (Q111) led to a switch in the enantiopreference of the ArM. X-ray crystal structures of SCP_Q111CBpy and SCP_Q111BpyAla ArMs with bound Cu(ii) ions unveiled crucial differences in the orientation of the catalytic centre. Combining structural information, alanine scanning studies, and computational analysis shed light on the distinct active sites of the ArMs, clarifying that these active sites stabilise the nucleophilic substrate on different sides of the electrophile leading to the observed switch in enantioselectivity. This work underscores the importance of integrating structural studies with catalytic screening to unravel the intricacies of ArM behaviour and facilitate their development for targeted applications in biocatalysis.

Ameliorative effect of Arabic gum Acacia and mori extracts in streptozotocin-induced diabetic rats: implications of Cas-3 and TGF-ß.

OBJECTIVE: Arabic gum Acacia (AG) is rich in fiber which improves lipid metabolism besides its antioxidant effect. Folium mori (FM) is a widely used herb due to its immunomodulatory, antimicrobial, and antioxidant activity. In the current study, we explore the antidiabetic, anti-inflammatory, as well as antioxidant activities of AG and FM in Streptozotocin (STZ), induced diabetic rats. MATERIALS AND METHODS: STZ diabetic rats were orally administrated with metformin and/or a combination of AG and FM for 4 weeks. Glycemic levels, Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), cholesterol, triglycerides, urea, and creatinine were determined. Malondialdehyde (MDA), glutathione peroxidase (GPx), and Superoxide dismutase (SOD) were also evaluated. Gene expression and profile as well as immunohistopathological were also evaluated. RESULTS: The results elicited no toxicological profile of both AG and FM. Plasma glucose level was decreased starting from 1st week to 4th week; besides, there was an improvement in glycated hemoglobin, insulin, and fructosamine. Liver and kidney damage markers were decreased in both AG and FM-treated rats. A significant increase in the antioxidant defense system and a decrease in oxidative stress markers were also observed. Gene expression analysis in brain tissues revealed a significant decrease in Interleukin beta 1 (IL-ß1), Caspase 3 (Cas-3), and Transforming growth factor beta (TGF-ß). CONCLUSIONS: Oral treatment of metformin with AG and FM in STZ-injected rats could ameliorate protective pathways and can be one of the promising oral anti-diabetic herbal agents.

The emerging role of lncRNA MEG3 and MEG3 rs7158663 in hepatocellular carcinoma.

OBJECTIVE: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy in Egypt. Genetic and environmental factors play a role in its development. This study explored the association between the long non-coding RNA (lncRNA) MEG3 rs7158663 polymorphism, MEG3 expression, and the risk of HCC and other clinicopathologic characteristics in an Egyptian population. PATIENTS AND METHODS: This case-control study included 114 patients with HCC and 110 healthy controls. TaqMan Real-time PCR was used to analyze lncRNA MEG3 rs7158663. Serum MEG3 expression levels were measured using RT-PCR. RESULTS: The AA, GA+AA, and A alleles were associated with increased risk for HCC (adjusted odds ratio (OR) 11.84%, 95% CI 4.07-34.45, p < 0.0001; adjusted OR 3.18, 95% CI 1.79-5.67, p < 0.0001; and adjusted OR 2.87, 95% CI 1.91-4.34, p < 0.0001, respectively). The mutant genotype and allele were linked to an increased risk in male patients and patients ≥ 50 years old. MEG3 serum expression level was downregulated in HCC patients. The rs7158663 G > A polymorphism and downregulated MEG3 were significantly associated with larger tumor size and advanced disease stage. CONCLUSIONS: MEG3 rs7158663 single nucleotide polymorphisms and downregulated lncRNA MEG3 were associated with HCC risk and may represent diagnostic and bad prognostic factors for HCC patients.

Facile Microemulsion Synthesis of Vanadium-Doped ZnO Nanoparticles to Analyze the Compositional, Optical, and Electronic Properties.

In this work, microemulsion method has been followed to synthesize vanadium-doped Zn1-xVxO (with x = 0.0, 0.02, 0.04, 0.06, 0.08, and 0.10) nanoparticles. The prepared samples are characterized by several techniques to investigate the structural, morphology, electronic, functional bonding, and optical properties. X-ray diffractometer (XRD) analysis confirms the wurtzite phase of the undoped and V-doped ZnO nanoparticles. Variation in the lattice parameters ensures the incorporation of vanadium in the lattice of ZnO. Scanning electron microscopy (SEM) shows that by increasing contents of V ions, the average particle size increases gradually. X-ray Absorption Near Edge Spectroscopy (XANES) at the V L3,2 edge, oxygen K-edge, and Zn L3,2 edge reveals the presence and effect of vanadium contents in the Zn host lattice. Furthermore, the existence of chemical bonding and functional groups are also asserted by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). UV⁻Visible analysis shows that by increasing V⁺ contents, a reduction up to 2.92 eV in the energy band gap is observed, which is probably due to an increase in the free electron concentration and change in the lattice parameters.

Pharmaceutical Consultation in UAE Community Pharmacies.

In recent years, the focus of pharmacists as traditional drug dispensers has shifted to more active and participative role in risk assessment, risk management, and other medication related consultation activities. Pharmacy profession is evolving steadily in the United Arab Emirates (UAE). Pharmacists in UAE are so much occupied in their administrative and managerial duties that dispensing is mostly attended to by pharmacy technicians. Pharmacist-led patient counseling is limited to the dosage and frequency of medications and rarely adverse reactions and drug interactions with other medications. Therefore we decided to perform quantitative questionnaires study to explore the role of pharmacist in patient counseling in UAE, the evaluation of pharmacist's opinion on patient counseling and the potential determinants of personal consultation. Results show the frequency and nature of inquiries received by pharmacist. Five to twenty inquires per month are received from patient, most of them related to drug prescription and dose recommendation. Thirty nine percent of pharmacists received inquiries from doctors, most of them related to the dose and mode of action. Ninty two percent of the pharmacists agreed that patient counseling is their professional responsibility. About 82% of pharmacists agreed that counseling will increase their sales and enhance the reputation of their pharmacies. Seventy percent of pharmacists mentioned that they need to undergo training for effective counseling while 46% of pharmacists felt that more staff in the pharmacies would have a positive influence on patient compliance to medication therapies and patient safety. The potential determinants of personal consultation show that 52% of participants trusted pharmacist and 55% considered the pharmacist as a friend. Forty eight percent of participants visited the pharmacy for medical recommendation while 30% for drug compounding, 72% agreed that pharmacist conducts full instruction while 31% agreed about full investigation. In conclusion, reorganization of the pharmacist's activities may improve pharmaceutical consultations. Pharmacists must be exposed to recent trends in drug therapy, dosage forms, dosage, adverse effects and interaction. This will go a long way in providing rational use of drugs to the patients and improve their quality of life.

Heroin profiling: mannitol hexaacetate as an unusual ingredient of some illicit drug seizures.

Mannitol hexaacetate (MHA) has been detected by GC-MS in some brown illicit drug seizures in which diacetylmorphine (DAM) was completely lacking. The presence of MHA as a genuine ingredient of the drug seizures rather than a storage- or an analytical artifact, formed by transacetylation of mannitol with the DAM in the heroin, has been verified. It is argued that MHA was formed as a result of the addition of mannitol, as diluent, before the acetylating step in the process of the heroin preparation. This early dilution in the production and distribution chain of the illicit drug is deemed as peculiar, nonetheless it may be highly indicative of a specific production process of heroin in a trafficking organization.

The use of cyclohexanone as a "derivatizing" reagent for the GC-MS detection of amphetamines and ephedrines in seizures and the urine.

A GC-MS method has been developed for the detection of amphetamine, methamphetamine, and the ephedrines, in seizures and the urine, based on on-GC condensation (derivatization) with cyclohexanone. The method is simple: the dried seizure material or the urine extract was mixed with cyclohexanone and injected into the GC-MS. The method was found to be superior to the methods based on acyl and trimethylsilyl (TMS) derivatization. Unlike for the acyl and TMS derivatives, the molecular and fragment ions of the cyclohexanone condensation products (cyclohexanone derivatives) were of substantial abundance, a useful property in unambiguous compound characterization. Furthermore, the high stability of the "derivatizing" reagent, cyclohexanone, compared with acyl and TMS derivatizing reagents, is a useful property in method development. The present method has proved selective and, tentatively, sensitive enough in the following areas (where methods based on acyl and TMS derivatization, as tested in this laboratory, have failed): (a) detection of amphetamine as a metabolite of methamphetamine; (b) detection of norpseudoephedrine as a metabolite of pseudoephedrine; (c) detection of amphetamine as an impurity of methamphetamine; (d) detection of cathine (norephedrine) as a constituent of Khat leaves; and (e) differentiation of Khat use from phenylpropanolamine use.

Increased bcl-2 expression is associated with primary resistance to chemotherapy in human epithelial ovarian cancer.

OBJECTIVE: bcl-2, an anti-apoptotic factor, has a role in the pathogenesis of ovarian cancer as well as in resistance to chemotherapy. DESIGN AND METHODS: 20 benign, and 26 malignant epithelial ovarian tissues were analyzed for bcl-2 protein and mutant p53 by enzyme-immunoassay (EIA). Flowcytometric analysis was also performed. Patients of malignant group were followed up to monitor overall survival and primary resistance to chemotherapy. RESULTS: bcl-2 was significantly higher in malignant group than benign group (p < 0.001). A cutoff value was determined for bcl-2 (63.8 kU/g protein). At this cutoff, sensitivity is 80.7%, and specificity is 85%. Using chi square analysis, a significant correlation was found between bcl-2 and FIGO stage (p = 0.01), overall survival (p = 0.01), as well as primary resistance to chemotherapy (p = 0.03). By correlation coefficient analysis the relation between bcl-2 and synthetic phase fraction was highly significant (p = 0.002). Bcl-2, p53, and FIGO stage were significantly correlated to poor survival (p = 0.01) in univariate analysis. However, in multivariate analysis, only FIGO stage, and p53 were independent risk factors. CONCLUSION: EIA could be a useful tool for investigating the prognostic value of bcl-2, and its possible prediction of platinum resistance in epithelial ovarian cancer. This might help in selecting patients for future anti-bcl-2 therapy.

Ectopic pregnancy presenting with obturator nerve pain.

A 27 year old woman had a three day history of pain in the cutaneous distribution of the left obturator nerve before she developed the classical picture of ectopic pregnancy with lower abdominal pain and vaginal bleeding. A left tubal pregnancy was subsequently confirmed by laparoscopy. Referred pain along the obturator nerve has been reported in other pelvic conditions, but has not previously been reported as a manifestation of ectopic pregnancy. Ectopic pregnancy may present with a very wide range of signs and symptoms and should be excluded in females of child bearing age with unexplained symptoms including pain anywhere from the shoulder down to the knee.