Band 36, Heft 3

Compared to Europe, data on genetic variation in genes transcribing drug metabolizing enzymes among Asian is limited due to ethnic diversity. Here we compare frequencies for clinically relevant single nucleotide polymorphism (SNP) commonly observed in drug metabolizing enzymes between European and Malaysian/Singaporean. Minor allele frequencies (MAF) for the indicated SNPs for European, South Asian and East Asian populations were obtained from the NCBI website (https://www.ncbi.nlm.nih.gov/snp). The SNP prevalence among Malaysian/Singaporean was characterized from gene association studies. Generally, some SNPs in CYP2D6 and CYP2C19 do not show good agreement between the two populations as to the MAF value obtained. CYP2D6*4 tends to be more common among European, whereas CYP2D6*10 is more common in Malays and Chinese among Singaporean. Regardless of different phenotype, MAF of CYP2D6*4 for Indians is similar to that seen by the European. Singaporeans show smaller MAF for CYP2C19*17 but higher CYP2C19*2 frequencies as opposed to European ones. Following growing attention to the contribution of CYP3A4/5, N-acetyltransferases ( NAT2 ), thiopurine methyltransferase ( TPMT ) and uridine diphosphate glucuronosyltransferases ( UGT ) 2B7 in predicting drug response across Europe, there are limited pharmacogenetics (PGx) studies examining the gene-drug interaction among Malaysian/Singaporean. To better understand the heterogeneity of the drug response, PGx studies for the abovementioned enzymes between ethnics in Malaysian/Singaporean should be identified.

Objectives To evaluate the association of VDR polymorphisms ( FokI , TaqI and ApaI ) with vitamin D levels and glycemic status in type 2 diabetes patients from Southern India. Methods In this observational study, genotype frequencies and vitamin D levels of 200 cases (type 2 diabetes patients) were compared with 300 controls (unrelated anonymised stored samples of healthy volunteers) from south India. Serum 25 (OH) D levels were measured by immunoassay technique, glycated hemoglobin (HbA 1c ) was measured using HPLC and genotyping of VDR polymorphisms were carried out using Real time Polymerase Chain Reaction (RT PCR). Results About 69.2% of type 2 diabetes patients were found to have vitamin D deficiency. FokI polymorphism showed variations in serum 25 (OH) D levels, with AA and AG genotypes having significantly lower serum 25 (OH) D levels as compared to GG [13.24 (8.4) ng/ml, 15.02 (7.07) ng/ml and 20.67 (13.64) ng/ml respectively]. There was no difference in HbA 1c levels with respect to the vitamin D levels and VDR polymorphisms. Conclusions AA and AG genotypes of FokI polymorphisms are associated with low serum 25 (OH) D levels. However there was no association between VDR polymorphisms and glycemic status in south Indian type 2 diabetes patients.

Objectives A database comprising multivariate data in developing a model from nuclear magnetic resonance (NMR) analysis using human bio fluids are necessary to have reproducibility and reliability of the data. To achieve reproducibility of the data, standardised experiments, including internal standard and preservative used should be attained, especially for samples such as human bio fluids to hinder the variation among samples. The aim of the study was to optimise in commonly used human bio fluids (serum and urine) for a suitable internal standard and preservative used in extended storage samples for NMR analysis. Methods Serum and urine samples were collected from healthy human subjects. The experiment was divided into two parts, part one to evaluate 2,2,2,2-tetradeutero-4,4-dimethyl-4-silapentanoic acid (TSP) and 4,4-dimethyl-4-silapentane-1-ammonium trifluoroacetate (DSA) as the optimal internal standard for the serum and urine samples. The second part investigated the effects of preservatives in the serum and urine samples on extended storage. Results Overall, TSP and DSA are suitable to be used as an internal standard in human urine samples. However, DSA is a superior internal standard in serum samples for NMR analysis. For the effect of preservative, the results indicated that human serum and urine samples could be stored without addition of preservative in −80 °C, as no changes in NMR fingerprinting have been observed during storage in the absence or presence of the preservative. Conclusions The findings suggest the use of DSA and TSP as an internal standard in serum and urine samples, respectively. Storage of serum and urine samples without any addition of preservative for an extended period has no effect on the metabolites changes. By having a standardised method, it will offer a considerable saving in both operator and spectrometer time and most importantly produce reproducible and reliable data.

Objectives In a previous investigation we searched for the occurrence of palmitic acid ester compounds of delta9-tetrahydrocannabinol (THC) and its primary metabolite 11-hydroxy-delta9-THC (11-OH-THC) in human body fluids and tissues (THC palmitic acid monoester [THC-Pal] and 11-OH-THC palmitic acid diester [11-OH-THC-DiPal]). As those esters could not be detected in various human body fluids (e.g. blood) or tissues (e.g. adipose tissue) we extended the investigation analyzing adipose tissue samples of mice previously given synthetic THC or a cannabis extract. Methods In total, 48 adipose tissue samples previously tested positive for THC by means of a liquid chromatographic triple quadrupole mass spectrometric (LC-QQQ-MS) method were analyzed for the presence of THC-Pal and 11-OH-THC-DiPal by means of LC-QQQ-MS. Results THC-Pal and 11-OH-THC-DiPal were not detected among the adipose tissue samples analyzed despite the presence of high THC concentrations within the adipose tissue. THC concentrations in adipose tissue were in a range of approximately 7–2,595 ng/g (median: 468 ng/g, average: 704 ng/g). Conclusions A (site-specific) synthesis of 11-OH-THC palmitic acid monoesters (11-hydroxy-delta9-THC-1-palmitate and 11-palmitoyloxy-delta9-THC) still remains to be done. After synthesis of these monoesters, their presence in the body fluids and tissues after THC administration should be investigated.

Objectives The function and expression of cytochrome P450 (CYP) drug metabolizing enzymes is highly variable, greatly affecting drug exposure, and therapeutic outcomes. The expression of these enzymes is known to be controlled by many transcription factors (TFs), including ligand-free estrogen receptor alpha (ESR1, in the absence of estrogen). However, the relationship between the expression of ESR1, other TFs, and CYP enzymes in human liver is still unclear. Methods Using real-time PCR, we quantified the mRNA levels of 12 CYP enzymes and nine TFs in 246 human liver samples from European American (EA, n = 133) and African American (AA, n = 113) donors. Results Our results showed higher expression levels of ESR1 and six CYP enzymes in EA than in AA. Partial least square regression analysis showed that ESR1 is the top-ranking TF associating with the expression of eight CYP enzymes, six of which showed racial difference in expression. Conversely, four CYP enzymes without racial difference in expression did not have ESR1 as a top-ranking TF. These results indicate that ESR1 may contribute to variation in CYP enzyme expression between these two ancestral backgrounds. Conclusions These results are consistent with our previous study showing ESR1 as a master regulator for the expression of several CYP enzymes. Therefore, factors affecting ESR1 expression may have broad influence on drug metabolism through altered expression of CYP enzymes.

Objectives The anti-inflammatory activity of Boswellia serrata extracts (BSE) is well known. BSE comprises boswellic acids (BA) such as 3- O -acetyl-11-keto- beta -boswellic acid (AKBA) and 11- keto -boswellic acid (KBA) as major constituents. One of the limitations of BAs is their poor oral bioavailability. The aim of the study was to prepare solid lipid particles of Boswellia serrata extract (SLBSP) to enhance the bioavailability of BAs. Methods The pharmacokinetic profile of BAs was studied in 10 healthy human volunteers following a single oral dose of 333 mg of SLBSP. Pharmacokinetic blood samples were collected at 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 8, and 12 h post drug administration. Plasma KBA and AKBA levels were measured using a validated LC-MS/MS method. Pharmacokinetics parameters were estimated using Pheonix WinNonlin (Build 6.4.0.768) software. Results Ten healthy human volunteers were included and peak plasma concentration was achieved in 1.5 and 2.3 h for AKBA and KBA respectively. Maximum plasma concentration ( C max ) was 8.04 ± 1.67 ng/mL for AKBA and 23.83 ± 4.41 ng/mL for KBA whereas the corresponding area under the concentration-time curve (AUC) was 136.7 ± 56.77 ng/mL*h and 165.7 ± 24.5 ng/mL*h respectively. The elimination half-life ( t 1/2 ) of AKBA and KBA was 6.8 ± 3.0 h and 2.45 ± 0.3 h respectively. Conclusions The SLBSP formulation of BSE showed enhanced oral bioavailability of BAs compared with historically reported data of unformulated BSE.

Objectives This study investigates protection against oxidative stress and memory enhancing potential of long-term consumption of Moringa oleifera leaves. Methods Male Wistar rat were fed with mixture of M. oleifera -supplemented diets (MOSD) partitioned in 1, 5, 10, and 20% continuously for 12 weeks. Object recognition test (ORT) and Morris water maze (MWM) was used for assessing neurocognition. Changes in body weight, Lipid peroxidation (MDA), Glutathione (GSH), Catalase (CAT) and Acetylcholinesterase (AChE) activity was assayed in the brain tissue. Histomorphometric of the hippocampus was also examined. Results The diets progressively increase the body weigh after the 12 weeks, improved spatial (MWM) and non-spatial (ORT) memory performance, protect against oxidative stress, inhibit AChE activity and suppresses neuronal degeneration in the hippocampus when stained with Cresyl violent stain. Conclusions Conclusively, long-term consumption of MOSD shows strong protection against oxidative stress and hippocampal degeneration and improves neurocognition with dose dependent effect.

Objectives Since melperone abuse with lethal intoxication is common, expert opinions based on therapeutical and lethal concentration ranges can be considered as important. Because there is a lack of information about fatalities caused by melperone mono-intoxications and data on tissue samples with concentration distribution, the aim of this work is the examination of lethal concentration ranges of melperone and drug quantification in different matrices. Methods An LC-MS/MS method was applied for analyses performed in blood and tissue samples. Quantification based on standard addition and sample preparation on liquid–liquid extraction with 1-chlorobutane. An appropriate tissue homogenization was performed ahead of extraction with an IKA Ultra-Turrax-Tube-Drive ® . A Luna 5 µm C18 (2) 100 Å, 150  × 2 mm analytical column was used for chromatographic separation and the elution was performed with two mobile phases consisted of A (H 2 O/methanol = 95/5, v/v) and B (H 2 O/methanol = 3/97, v/v) both with 10 mM ammonium acetate and 0.1% acetic acid. Results A multi-drug LC-MS/MS analytical method developed was applied successfully for melperone quantification in different post-mortem matrices. No analytical problems could be identified during method development and analyses of real samples. The melperone lethal concentration calculated in femoral blood of the drug mono-intoxication investigated was 10 mg/L. Melperone concentration distribution was presented for the first time. Conclusions The lethal reference concentration of melperone in femoral blood of 17.1 mg/L pointed out in different reference lists should be used with caution. Instead, a lower lethal melperone concentration should be considered. The post-mortem concentration distribution of the drug presented could be helpful in the interpretation of cases where no blood samples are available.

Objectives Osteoporosis often occurs in individuals of different age groups, frequently during menopause and after ovariectomy. It increases the risk of pathological fractures almost twice. The aim of our research was to assess bone metabolism, nanocomposite structure of the tibia under conditions of ovariectomy and zoledronic acid treatment. Methods X-ray diffraction has been performed for nanostructure analysis of mineral crystallites and crystal lattice of hydroxyapatite in the tibia samples of ovariectomized rats with additional application of bisphosphonate zoledronic acid (0.025 mg/kg). Markers of remodeling – osteocalcin, alkaline phosphatase, tartrate resistant acid phosphatase 5b – were determined. Quantitative amount of calcium in the bones was detected by atomic absorption method. Results Zoledronic acid prevented loss of mineral mass after ovariectomy. Rats after ovariectomy, treated with zoledronic acid, showed statistically higher (р<0.05) values of crystalline phase and calcium content compared with the SHAM-surgery and ovariectomy groups (р<0.05). Zoledronic acid inhibited bone remodeling, which is proved by tartrate resistant acid phosphatase 5b reduction and inhibition of osteoclasts during the experiment. Conclusions These results enable to suggest that zoledronic acid can improve mineral mass of the bone during menopause in individuals of different age groups.

Objectives Pain in joints is a major clinical problem since ancient times. Waja-uz-Zahr stands for low back pain which may arise from internal or external structures of lumbosacral region due to Su-e-Mizaj sada or maddi due to the surplus of Burudat or accumulation of Kham Balgham in lower back. Imala (diversion) and Taqiya-e-Mavad (Evacuation) are inevitable in case of maddi amraz . Hijama (cupping therapy) is a simple, safe, tolerable, and effective regimenal modality, helpful in the treatment of aches and pains. Gruner mentioned a special kind of cupping method in the canon of medicine of Avicenna where cupping glasses are glided over a specified area of body intending the diversion of morbid material under the name of Bier’s treatment. The purpose of present case series was to provide the insight into the effect of gliding cupping in low back ache. Case presentation Three patients of Waja-uz-Zahr , aged between 29 and 64 years, were given gliding cupping treatment after the application of Roghan-e-Surkh on lumbosacral region alternatively for 14 days. Oswestry disability index (ODI) and visual analogue scale (VAS) were used for the assessment of efficacy. Improvement, in case 1, case 2, and case 3 in ODI score was 42.42, 68.18, and 62.50%, respectively while in VAS score, it was 50, 62.5, and 50%, respectively, was observed without any adverse effect. Conclusions On the basis of the above results, it can be concluded that the gliding cupping therapy may be used as an effective modality of treatment for low back ache.