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B. C. Behera, Pune, India
T. Chen, Stanford, CA/USA
E. Corsini, Milan, Italy
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M. Lotti, Bonn, Germany / Padova, Italy
P. Micke, Uppsala, Sweden
A. N. Misra, Ranchi, Jharkhand State, India
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K. Renganathan, Johnstown, PA/USA
S. D. Ray, Fort Wayne, IN/USA
M. Schwarz, Tuebingen, Germany
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A. Winterpacht, Erlangen, Germany
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Now showing 1 - 20 of 71

Development of competitive lateral flow immunoassay coupled with silver enhancement for simple and sensitive salivary cortisol detection
(2018-12-21) Apilux, Amara; Rengpipat, Sirirat; Suwanjang, Wilasinee; Chailapakul, Orawon
Cortisol is known as a stress biomarker. The measurement of cortisol levels is an early warning indicator for health conditions and diagnosis of stress-related diseases. Herein, a lateral flow immunoassay using a gold nanoparticle label with a silver enhancement system was developed for the simple, sensitive and rapid detection of cortisol. The developed assay was based on a competitive platform of which cortisol-BSA conjugate was immobilized at the test zone to compete with an analyte. The quantitative analysis was performed using gold nanoparticles (AuNPs) as signal labeling. Sequentially, the silver enhancement solution was applied in order to enhance the sensitivity of the assay with the results easily seen by the naked eye. Using this system, the limit of detection (LOD) was found to be 0.5 ng/mL with a 3.6 fold more sensitive detection than without the enhancement system (LOD = 1.8 ng/mL). The salivary cortisol analysis was in the range of 0.5-150 ng/mL (R2 = 0.9984), which is in the clinical acceptable range. For the semi-quantitative analysis, the intensity color of the results was analyzed using an image processing program. The proposed method was successfully applied to detect cortisol in saliva. In addition, the results from our method also complied with the ones of those obtained by using the commercial enzyme-linked immunosorbent assay (ELISA). This developed assay offers great promise for a non-invasive screening test of salivary cortisol.
A haplotype of the phosphodiesterase 4D (PDE4D) gene is associated with myocardial infarction and with cardiometabolic parameters
(2018-12-19) Rodríguez-Pérez, José Manuel; Posadas-Sánchez, Rosalinda; Blachman-Braun, Ruben; Vargas-Alarcón, Gilberto; Posadas-Romero, Carlos; García-Flores, Esbeidy; López-Bautista, Fabiola; Tovilla-Zárate, Carlos Alfonso; González-Castro, Thelma Beatriz; Borgonio-Cuadra, Verónica Marusa; Pérez-Hernández, Nonanzit
The phosphodiesterase family is involved in a wide spectrum of diseases, including ischemic stroke. However, few studies have analyzed the relationship between phosphodiesterase 4D (PDE4D) and myocardial infarction (MI). Therefore, the aim of this research was to evaluate the association of the PDE4D gene polymorphisms with MI, and with cardiometabolic parameters in the Mexican population. Six polymorphisms (rs2910829, rs1423246, rs966221, rs4502776, rs13172481, and rs6869495) were genotyped in 1023 MI patients and 1105 healthy controls. A similar distribution of the six polymorphisms was observed in both studied groups. However, after evaluating the linkage disequilibrium, we detected a risk haplotype for MI (AGAGAA; OR = 1.148; P = 0.025). In addition, the polymorphisms were associated with the presence of some clinical and metabolic parameters (central obesity, hypertriglyceridemia, Aspartate transaminase >p75, Lipoprotein (a) >30 mg/dL, TAT >p75, fatty liver, and vitamin D <30 ng/dL) in healthy controls. The results suggest that in the Mexican population, a PDE4D haplotype is associated with increased risk of developing MI, and that PDE4D polymorphisms are independently associated with the presence of cardiometabolic parameters.
Androgen receptor (AR)-CAG trinucleotide repeat length and idiopathic male infertility
(2018-12-17) Mobasseri, Narges; Babaei, Faezeh; Karimian, Mohammad; Nikzad, Hossein
CAG trinucleotide repeats in androgen receptor (AR) gene encode a polyglutamine tract in AR N-terminal transactivation domain. Studies have been conducted to evaluate the effect of CAG repeat length on male infertility, which have yielded contradictory results. This study aimed to explore the number of AR-CAG repeats in 150 fertile controls and 150 idiopathic infertile men, divided into four azoospermia, oligozoospermia, asthenozoospermia, and teratozoospermia subgroups. In addition, a meta-analysis was conducted based on previous studies to assess the association of the mentioned variation with male infertility in recent years. Polymerase chain reaction (PCR) targeting followed by an electrophoresis on polyacrylamide gel was used for AR-CAG genotype detecting. Moreover, a systematic search was performed in PubMed, Web of Science, Science Direct, and Google Scholar databases to collect eligible studies for meta-analysis purpose. According to the results, a significant association was observed between increased length of AR-CAG polymorphism and male infertility (p< 0.0001). Furthermore, there were similar significant associations in the azoospermia (p= 0.048), asthenozoospermia (p= 0.013) and teratozoospermia (p= 0.002) subgroups. In addition, meta-analysis on forty studies showed a significant association between AR-CAG polymorphism in the overall analysis (SMD= 0.199, 95 % CI= 0.112-0.287, p<0.001) and the Caucasian subgroup (SMD= 0.151, 95 % CI= 0.040-0.263, p= 0.008). Our results elucidated that long stretches of CAG repeat might lead to AR dysfunction, contributing to male infertility especially in the Caucasian population.
Protein arginine methyltransferase 5 promotes bladder cancer growth through inhibiting NF-kB dependent apoptosis
(2018-11-20) Hu, Guodong; Wang, Xiu; Han, Yi; Wang, Ping
Protein arginine methyltransferase 5 (PRMT5) has emerged as a key regulator of tumorigenesis. However, how PRMT5 functions in bladder cancer, the most common malignancy of the urological system, is unknown. We described here that PRMT5 is highly expressed in bladder cancer cell lines and primary human bladder cancer tissues. PRMT5 enhances the proliferation and colony formation of bladder cancer cells. PRMT5 knockdown induces bladder cancer cell apoptosis. Mechanistically, PRMT5 enhances NF-kB activation by targeting crucial anti-apoptotic genes such as BCLXL and c-IAP1, thereby inhibiting tumor cell apoptosis and sustaining proliferation. Importantly, PRMT5 inhibitor opposed tumor growth and BCLXL and c-IAP1 transcription in the bladder cancer xenograft model. Collectively, the current suggests the crucial role of PRMT5 as a promising therapeutic target in bladder cancers.
Cyclosporin A attenuating morphine tolerance through inhibiting NO/ERK signaling pathway in human glioblastoma cell line
(2018-11-12) Rashki, Asma; Mumtaz, Faiza; Jazayeri, Farahnaz; Shadboorestan, Amir; Esmaeili, Jamileh; Ejtemaei Mehr, Shahram; Ghahremani, Mohammad Hossein; Dehpour, Ahmad Reza
Cyclosporin A (CsA) is known to have an immunosuppressive action. However, it is also attracting attention due to its effects on the nervous system, such as inhibiting the development and expression of morphine-induced tolerance and dependence through unknown mechanisms. It has been shown that CsA modulates the nitric oxide (NO) synthesis and extracellular signal-regulated kinases (ERK) activation, which are potentially involved in signaling pathways in morphine-induced tolerance in cellular models. Therefore, the current study was designed to evaluate the modulatory role of CsA on the MOR tolerance, by targeting the downstream signaling pathway of NO and ERK using an in vitro model. For this purpose, T98G cells were pretreated with CsA, calcineurin autoinhibitory peptide (CAIP), and NG-nitro-l-arginine methyl ester (L-NAME) 30 min before 18 h exposure to MOR. Then, we analyzed the intracellular cyclic adenosine monophosphate (cAMP) levels and also the expression of phosphorylated ERK and nitric oxide synthase (nNOS) proteins. Our results showed that CsA (1 nM, 10 nM, and 100 nM) and CAIP (50 μM) have significantly reduced cAMP and nitrite levels as compared to MOR-treated (2.5 μM) T98G cells. This clearly revealed the attenuation of MOR tolerance by CsA. The expression of nNOS and p-ERK proteins were down-regulated when the T98G cells were pretreated with CsA (1 nM, 10 nM, and 100 nM), CAIP (50 μM), and L-NAME (0.1 mM) as compared to MOR. In conclusion, the CsA pretreatment had a modulatory role in MOR-induced tolerance, which was possibly mediated through NO/ERK signaling pathway.
Differential pattern of brain functional connectome in obsessive-compulsive disorder versus healthy controls
(2018-11-08) Yazdi-Ravandi, Saeid; Akhavanpour, Hassan; Shamsaei, Farshid; Matinnia, Nasrin; Ahmadpanah, Mohammad; Ghaleiha, Ali; Khosrowabadi, Reza
Researchers believe that recognition of functional impairment in some of brain networks such as frontal-parietal, default mode network (DMN), anterior medial prefrontal cortex (MPFC) and striatal structures could be a beneficial biomarker for diagnosis of obsessive-compulsive disorder (OCD). Although it is well recognized brain functional connectome in OCD patients shows changes, debate still remains on characteristics of the changes. In this regard, little has been done so far to statistically assess the altered pattern using whole brain electroencephalography. In this study, resting state EEG data of 39 outpatients with OCD and 19 healthy controls (HC) were recorded. After, brain functional network was estimated from the cleaned EEG data using the weighted phase lag index algorithm. Output matrices of OCD group and HCs were then statistically compared to represent meaningful differences. Significant differences in functional connectivity pattern were demonstrated in several regions. As expected the most significant changes were observed in frontal cortex, more significant in frontal-temporal connections (between F3 and F7, and T5 regions). These results in OCD patients are consistent with previous studies and confirm the role of frontal and temporal brain regions in OCD.
A meta-analysis of the relationship between body mass index and risk of rheumatoid arthritis
(2018-11-06) Zhou, Ying; Sun, Mingfang
The present meta-analysis aimed to evaluate the relationship between body mass index (BMI) and rheumatoid arthritis (RA). A systematic search of the Cochrane, Pubmed, and Embase databases was conducted to identify relevant studies published before September 2017 using terms related to BMI and RA. Fixed or random-effect models were used to estimate the pooled relative risk (RR) with 95 % confidence interval (CI). Subgroup analyses by sex were performed to investigate the association between BMI and RA in male and female subgroups. A total of 14 eligible studies containing 353,948 patients were included in the analysis. The pooled results suggested that the odds ratios (ORs) of RA were 1.08 (95 % CI: 1.00~1.15) for overweight, and 1.32 (95 % CI: 1.11~1.54) for obesity, respectively, suggesting that a higher BMI increases the risk of RA compared to normal weight. Further subgroup analyses showed a positive association between BMI and RA risk but only in females, with a RR of 1.11 (95 % CI: 1.00~1.22) for overweight and 1.40 (95 % CI: 1.24~1.57) for obesity. In conclusion, an increased BMI may lead to a higher risk for RA development. Furthermore, the positive association between BMI and RA risk may be stronger in female populations than in males. However, additional analyses are needed.
Read across for the derivation of Indoor Air Guidance Values supported by PBTK modelling
(2018-11-05) Schupp, Thomas
Polyurethane Flexible Foams (PUF) are versatile materials used in upholstered furniture and bed mattresses. Due to the production procedure, fresh foams emit volatile organic compounds (VOC) which may contribute to indoor air exposure. To evaluate the risk for consumers, the VOC concentration measured in chamber tests can be matched against existing benchmarks for indoor air like “Richtwerte” (RW) of the German UBA (Umweltbundesamt), “Lowest Concentration of Interest” (LCI) for construction products or derived no effect levels (DNEL) for consumer inhalation exposure. In a previous paper a method for the derivation of Indoor Air Guidance Values (IAGV) for VOC without RW, LCI or DNEL was developed. The method described made use of a sufficient toxicological database. For substances with an insufficient database, read across to structural analogues is a way forward to estimate Indoor Air Guidance Values (IAGV). In this work a read across exercise, supported by an open source physiology based toxicokinetic (PBTK) modelling program is demonstrated. The use of enzyme kinetic data for phase I and phase II metabolism is discussed and areas for further work were identified. For two substances with very limited toxicological data, allyloxypropanol (isomer mixture of 1-allyloxy-2-propanol and 2-allyloxy-1-propanol) and 2,3-di-ethyl-2,3-dimethylsuccinodintrile, Tentative Indoor Air Guidance Values of 750 μg/m³ and 65 μg/m³ were derived.
Immunogenicity of a novel tetravalent dengue envelope protein domain III-based antigen in mice
(2018-11-05) Fahimi, Hossein; Sadeghizadeh, Majid; Hassan, Zuhair M.; Auerswald, Heidi; Schreiber, Michael
Dengue virus is a mosquito-borne pathogen that causes dengue diseases. All four serotypes of dengue virus are infectious for humans. Therefore, an efficacious dengue vaccine should be tetravalent to provide protection against all types of virus. The goal of this study was to design a new tetravalent recombinant protein from envelope protein of dengue viruses to induce virus-neutralizing antibodies against all four serotypes in mice. A chimeric protein was designed from domain III of envelope protein of all serotypes of dengue virus. Four domain III fragments were linked together by alpha helix making linkers. The final sequence of the designed protein was analyzed in silico and the coding gene sequence was deduced by reverse translation. After cloning and expression of the recombinant protein (ED3-tetravalent protein), identity of the purified protein was confirmed using a pan-dengue specific monoclonal antibody in Western blotting. Then, the immunogenicity of the purified protein was studied in mice using antibody titration. The efficacy of induced antibodies in neutralization of the virus was studies by FRNT method. Furthermore, the induction of cellular immunity was studied by measurement of cytokines using ELISA method and measurement of lymphocyte proliferation using MTT assay. The ED3-tetravalent protein was able to enhance neutralizing immunogenic response against all four dengue serotypes; in similar way to that of tetravalent formulation of four individual domain III-based polypeptides. It is suggested that the ED3-tetravalent fusion protein can induce broadly neutralizing antibody responses against all four serotypes of dengue virus in mice.
Study of correlation between the NAT2 phenotype and genotype status among Greenlandic Inuit
(2018-11-02) Birch Kristensen, Emilie; Yakimov, Victor; Bjorn-Mortensen, Karen; Soborg, Bolette; Koch, Anders; Andersson, Mikael; Birch Kristensen, Kasper; Michelsen, Sascha Wilk; Skotte, Line; Ahrendt Bjerregaard, Anne; Blaszkewicz, Meinolf; Golka, Klaus; Hengstler, Jan G.; Feenstra, Bjarke; Melbye, Mads; Geller, Frank
N-acetyltransferase 2 (NAT2) is the main enzyme metabolizing isoniazid and genotype-based treatment has been studied for years without becoming common practice. To investigate whether genotype-based isoniazid treatment is feasible in Greenland, we sequenced the coding sequence of NAT2 and determined the NAT2 enzyme-activity by caffeine test. No additional genetic variants were identified in the coding sequence of NAT2, so that genotype status in 260 study participants could be assessed by a well-established 7-SNP panel. Studying the enzyme activity by the ratio of the two caffeine metabolites AFMU and 1X in 260 participants showed a high rate of slow phenotypes with intermediate or rapid genotype. These misclassifications were mainly observed in urine samples with pH<3, a deviation from the standard protocol due to the field work character of the study, where immediate pH adjustment to pH=3.5 was not possible. We excluded these samples. For the remaining 143 individuals with pH>3, we observed a moderate level of discrepancies (19 of the 116 individuals with intermediate or rapid genotype status having a slow phenotype). Further investigation showed that drinking coffee and not tea or cola was the most important factor for high levels of both metabolites. The concordance between phenotype and genotype status with regard to slow metabolism supported the recommendation of lower isoniazid doses in individuals with slow genotype status in order to avoid liver injury, a frequent side effect. The phenotypical variation observed for individuals with intermediate or rapid genotype status warrants further research before increased dosing of isoniazid can be recommended.
The inflammatory cytokine TNF contributes with RAC3-induced malignant transformation
(2018-11-02) Machado, Mileni Soares; Rosa, Francisco D.; Lira, María C.; Urtreger, Alejandro J.; Rubio, María F.; Costas, Mónica A.
RAC3 is a coactivator of steroid receptors and NF-kB. It is usually overexpressed in several tumors, contributes to maintain cancer stem cells and also to induce them when is overexpressed in non-tumoral cells. In this work, we investigated whether the inflammatory cytokine TNF may contribute to the transforming effects of RAC3 overexpression in the non-tumoral HEK293 cell line. The study model included the HEK293 tumoral transformed cell line constitutively overexpressing RAC3 by stable transfection and control non-tumoral cells transfected with an empty vector. The HeLa and T47D tumoral cells that naturally overexpress RAC3 were used as positive control. We found that TNF potentiated RAC3-induced mesenchymal transition, involving an increased E-Cadherin downregulation, Vimentin and SNAIL upregulation and enhanced migratory behavior. Moreover, concerning the molecular mechanisms by which TNF potentiates the RAC3 transforming action, they involve the IKK activation, which in addition induced the -Catenin transactivation. Our results demonstrate that although RAC3 overexpression could be a signal strong enough to induce cancer stem cells, the inflammatory microenvironment may be playing a key role contributing to the migratory and invasive phenotype required for metastasis and cancer persistence.
Comparing task performance, visual comfort and alertness under different lighting sources
(2018-10-29) Kazemi, Reza; Choobineh, Alireza; Taheri, Shirin; Rastipishe, Pegah
The aim of this study is to compare the effects of different light sources – namely light-emitting diode (LED), compact fluorescent (FLcomp) and fluorescent with warm color temperature (FLwarm) and cool color temperature (FLcool) – on the performances, alertness, visual comfort level and preferences in a pilot study. A laboratory controlled experiment was conducted by focusing on 20 postgraduate students who volunteered to participate in a series of tests under four different light sources. “GO NO GO” task and Karolinska Sleepiness Scale (KSS) were employed to assess objective and subjective alertness, while modified OLS questionnaire was used to gauge comfort level and preferences. In addition, editing and typing tasks were carried out as a performance evaluation. Significant increase was observed in subjective and objective alertness level under FLcool condition and LED in comparison to FLwarm and FLcomp (p < 0.05). In terms of typing performances, respondents performed significantly better with regard to typing speed under FLcool than FLwarm and FLcomp. The lowest number of typing errors was made under FLcool, followed by LED, FLcomp and FLwarm. LED was the most preferred (p=0.001) and most comfortable (p=0.011) lighting condition. The study concludes that the FLcool and LED were more beneficial for alertness level and performance for both computer-based and paper-based activities.
Everolimus, a mammalian target of rapamycin inhibitor, ameliorated streptozotocin-induced learning and memory deficits via neurochemical alterations in male rats
(2018-10-29) Fanoudi, Sahar; Hosseini, Mahmoud; Alavi, Mohaddeseh Sadat; Boroushaki, Mohammad Taher; Hosseini, Azar; Sadeghnia, Hamid R.
Everolimus (EVR), as a rapamycin analog, is a selective inhibitor of the mammalian target of rapamycin (mTOR) kinase and its associated signaling pathway. mTOR is a serine/threonine protein kinase and its hyperactivity is involved in the pathophysiology of Alzheimer’s disease (AD) and associated cognitive deficits. The present study evaluated the impact of EVR, on cognitive functions, hippocampal cell loss, and neurochemical parameters in the intracerebroventricular streptozotocin (icv-STZ) model of AD rats. EVR (1 and 5 mg/kg) was administered for 21 days following the single administration of STZ (3 mg/kg, icv) or for 7 days on days 21-28 post-STZ injection after establishment of cognitive dysfunction. Cognitive deficits (passive avoidance and spatial memory), oxidative stress parameters, acetylcholinesterase (AChE) activity, and percentage of cell loss were evaluated in the hippocampus. Chronic administration (1 and 5 mg/kg for 21 days from the day of surgery and icv-STZ infusion) or acute injection (5 mg/kg for 7 days after establishment of cognitive impairment) of EVR significantly attenuated cognitive dysfunction, neuronal loss, oxidative stress and AChE activity in the hippocampus of STZ-AD rats. In conclusion, our study showed that EVR could prevent or improve deteriorations in behavioral, biochemical and histopathological features of the icv-STZ rat model of AD. Therefore, inhibition of the hyperactivated mTOR may be an important therapeutic target for AD.
An automated method for the evaluation of breast cancer using infrared thermography
(2018-10-26) Morales-Cervantes, Antony; Kolosovas-Machuca, Eleazar Samuel; Guevara, Edgar; Maruris Reducindo, Mireya; Bello Hernández, Alix Berenice; Ramos García, Manuel; González, Francisco Javier
Breast cancer is one of the major causes of death for women. Temperature measurement is advantageous because it is non-invasive, non-destructive, and cost-effective. Temperature measurement through infrared thermography is useful to detect changes in blood perfusion that can occur due to inflammation, angiogenesis, or other pathological causes. In this work, we analyzed 206 thermograms of patients with suspected breast cancer, using a classification method, in which thermal asymmetries were computed, the most vascularized areas of each breast were extracted and compared; then these two metrics were added to yield a thermal score, indicative of thermal anomalies. The classification method based on this thermal score allowed us to obtain the test sensitivity of 100 %, specificity of 68.68 %; a positive predictive value of 11.42 % and negative predictive value of 100 %. These results highlight the potential of thermography imaging as adjunctive tool to mammography in breast cancer screening.
Risk factors and prevalence of toxocariasis in pregnant women and diabetic patients compared to healthy adults in Ilam province, western Iran
(2018-10-23) Raissi, Vahid; Sohrabi, Zahra; Getso, Muhammad; Raiesi, Omid; Hashemi Hafshejani, Saeideh; Shabandoust, Hajar; Etemadi, Soudabeh
Toxocara is one of the common intestinal nematodes in dogs and cats and is the agent of tissue migratory larvae in humans. Customarily, the prevalence of human toxocariasis hovers around 15.8 % in Iran. Furthermore, other research outcomes demonstrated a tendency for an outbreak of toxocariasis in Iran. Therefore, we carried out a cross-sectional study and assessed the seroprevalence of toxocariasis humans in Ilam Province, western of Iran. A total of 539 serum samples were collected between September 2017 and March 2018 from patients referred to the Health Centers of Ilam province, Iran. Serum samples were investigated for the presence of Toxocara using IgG antibodies, ELISA (Enzyme-Linked Immunosorbent Assay) kit. Risk factors such as contact with cats and dogs, living in rural areas were investigated among the study population. Out of 539 total samples collected, 97 cases (17.99 %) were positive for anti-toxocara IgG antibodies. These antibodies were recovered from serum samples of otherwise healthy adults (15.54 %, 49/296), pregnant women (21.16 %, 40/189) and diabetic patients (14.81 %, 8/54). This study showed significant relationship between toxocariasis and contact with animal pets in all studied groups (P value ≤ 0.05) and a significant relationship between toxocariasis and living in rural areas among pregnant women (P value ≤ 0.05).
Simultaneous determination of sarcosine and its related metabolites by gas chromatography-tandem mass spectrometry for prostate cancer diagnosis
(2018-10-16) Yamkamon, Vichanan; Yee, Pyone Pyone; Yainoi, Sakda; Eiamphungporn, Warawan; Suksrichavalit, Thummaruk
Shortly after sarcosine was delineated as a potential biomarker for prostate cancer in 2009, a variety of analytical methods for clinical application were developed. Moreover, higher uptake of glycine in the mitochondria also played a role in cancer proliferation. A major constraint in the accurate quantification of sarcosine was the interference of the two isomers, α-alanine and β-alanine, using chromatographic separation techniques. Accordingly, we aimed to develop an analytical method for determining sarcosine and its related metabolites (α- and β-alanine, glycine and creatinine) under the same conditions by gas chromatography-tandem mass spectrometry (GCMS/ MS). BSTFA + 1 % TMCS was used for silylation, and GC-MS/MS conditions were optimized for the target analytes. The unique transition ions of sarcosine, α- and β-alanine, glycine and creatinine set up in MRM acquisition were m/z 116 → 73, 190 → 147, 176 → 147, 176 → 147 and 100 → 73, respectively. This newly developed method was successfully validated to apply in clinical settings with low limits of detection (0.01 - 0.03 μg•mL-1), high correlations (R2 > 0.99), great accuracy (88 – 110 % recovery), and notable precision (RSD < 10 %). All TMS derivatives were > 80 % stable for up to 2 h after derivatization and analyzing during this period promises to achieve an accurate result. Monitoring the five-substance profile could enhance prospects for early diagnosis of prostate cancer.
Determination of tricyclic antidepressants in human urine samples by the three-step sample pretreatment followed by HPLC-UV analysis
(2018-09-24) Mohebbi, Ali; Farajzadeh, Mir Ali; Yaripour, Saeid; Afshar Mogaddam, Mohammad Reza
In this work, an efficient sample pretreatment method has been developed by combining salt induced– homogenous liquid–liquid extraction, dispersive solid phase extraction, and dispersive liquid–liquid microextraction based on the solidification of floating organic droplet for the extraction of some widely used tricyclic antidepressant (TCA) drugs (nortriptyline, amitriptyline, desipramine, clomipramine, and imipramine) in human urine samples before their determination by high performance liquid chromatography–ultraviolet detection. In brief, the target analytes are first isolated from urine samples into acetonitrile (ACN) separated by adding a salt. Then the obtained ACN phase is treated with a mixture of appropriate sorbents to remove interferences. Afterward, the purified ACN is mixed with menthol as an extractant and rapidly injected into alkaline HPLC–grade water as a preconcentration step. Next, the obtained solution is placed in an ice bath and menthol collects on top of the solution after solidification. The solidified drop is then withdrawn and injected into separation system after dissolving in 10 μL ACN. Under the optimum experimental conditions, extraction recoveries and enrichment factors of the selected drugs ranged from 69–84 % and 345–420, respectively. The limits of detection and quantification were obtained at the ranges of 0.22–0.31, and 0.71–1.1 μg L–1, respectively. The relative standard deviations of the proposed method were ≤ 6 % for intra– (n=6) and inter–day (n=4) precisions at a concentration of 10 μg L–1 (each drug). Finally, the suggested approach was applied to determine of TCA drugs in different patients' urine samples. The method could be applied in further TCAs pharmacokinetic and forensic studies.
Evaluation of microsatellite instability in tumor and tumor marginal samples of sporadic colorectal cancer using mononucleotide markers
(2018-09-24) Nouri Nojadeh, Jafar; Hashemzadeh, Shahriar; Samadi Kafil, Hossein; Behrouz Sharif, Shahin; Eftekharsadat, Amirtaher; Ghasemnejad, Tohid; Ghojazadeh, Mortaza; Sakhinia, Ebrahim
Microsatellite instability (MSI) is a unique molecular alteration that is due to a defective DNA mismatch repair (MMR) system. Approximately, 15-20 % of sporadic colorectal cancers (CRC) display MSI. Determination of MSI status in CRC has prognostic and predictive implications. Additionally, detecting MSI is used diagnostically for tumor detection and classification. The present study analyzed a panel of five mononucleotide markers, BAT- 25, BAT-26, NR-21, NR-22 and NR-27, amplified in a single multiplex PCR reaction to evaluate MSI status in CRC patients. Genomic DNA from 50 CRC and paired adjacent normal tissues was used for PCR-based MSI analysis. Our finding showed microsatellite instability in 36 % of specimens. Instability with differences in allele lengths was observed in the tumoral DNA compared to the tumor-free margin DNA sample. The frequency of instability in NR-21, BAT-26 and BAT-25 markers were more than others; their frequency were 35.48 %, 29.03 %, and 22.58 %, respectively. In conclusion, the NR-21, BAT-26, and BAT-25 were the most useful markers for discriminating cancer tissue from normal, therefore these markers have demonstrated promising potential for determining MSI status in patients with sporadic colorectal cancer.
The role of CA1 CB1 receptors on lithium-induced spatial memory impairment in rats
(2018-09-20) Vaseghi, Salar; Babapour, Vahab; Nasehi, Mohammad; Zarrindast, Mohammad-Reza
Lithium, a glycogen synthase kinase-3β (GSK-3β) inhibitor, prevents cannabinoid withdrawal syndrome, but there is limited data exploring the interaction between lithium and cannabinoid system on memory processes. The present study aimed to test the interaction between dorsal hippocampal (CA1 region) cannabinoid system and lithium on spatial memory in rats. Spatial memory was assessed in Morris Water Maze (MWM) apparatus by a single training session of eight trials. The results showed that pre-training intra-CA1 microinjection of ACPA, the cannabinoid type 1 receptor (CB1r) agonist, at doses of 0.001, 0.01 or 1 μg/rat, or AM251, the cannabinoid type 1 receptor (CB1r) antagonist, at doses of 1, 10 or 100 ng/rat, increased escape latency and traveled distance to the platform, suggesting a spatial learning impairment, whereas intraperitoneal administration of lithium (0.5, 1 or 5 mg/kg) had no effect on spatial learning. Also, rats that received lithium plus a lower dose of ACPA (0.001 μg/rat) or AM251 (1 ng/rat) had successful performance in the MWM. In the probe test, the results showed that pretraining administration of lithium (5 mg/kg) and ACPA (0.01 or 1 μg/rat) but not AM251 (at all doses used) impaired spatial memory retrieval. Also, lower dose of ACPA (0.001 μg/rat) or AM251 (1 ng/rat) potentiated the effect of ineffective doses of lithium (0.5 and 1 mg/kg) on spatial memory retrieval, while restored the effect of effective dose of lithium (5 mg/kg). In conclusion, cannabinoids may have a dual effect on lithium-induced spatial memory impairment in rats.
In vivo acute toxicity of detoxified Fuzi (lateral root of Aconitum carmichaeli) after a traditional detoxification process
(2019-08-31) Sun, Wan; Yan, Bo; Wang, Rongrong; Liu, Fucun; Hu, Zhengyan; Zhou, Li; Yan, Li; Zhou, Kang; Huang, Jiawei; Tong, Peijian; Shan, Letian; Efferth, Thomas
Many herbs of traditional Chinese medicine (TCM) possess not only therapeutic efficacy, but also toxicity towards normal tissues. The herbal toxicities occasionally cause serious adverse events or even fatal poisoning due to the erroneous use of TCM herbs. Fuzi (lateral root of Aconitum carmichaeli) is such an herb with its toxic ingredient, aconites. Aconitine, mesaconitine, and hypaconitine are the main toxic components of Fuzi, which are hydrolyzed into non-toxic derivatives by water decoction. Therefore, long-time decoction was commonly applied as a traditional way to detoxify Fuzi before use. Nevertheless, recent clinical trials presorted on adverse events induced by long-time decocted Fuzi, putting some doubt on the safety of Fuzi after the traditional detoxification procedure. To thoroughly determine whether or not long-time decocted Fuzi was safe, we conducted in vivo acute toxicity assays using both rodent and zebrafish models and performed chemoprofile analyses using HPLC and UPLC-MS. The HPLC analysis showed that toxic aconitine components were hydrolyzed into benzoyl derivatives with increasing time of decoction. These aconitines were undetected by HPLC in Fuzi after 2 hdecoction (FZ-120), indicating seemingly non-toxicity of FZ-120. Unlike the non-decocted Fuzi (FZ-0) and 60 min-decocted Fuzi (FZ-60) with lethal toxicity, FZ-120 at 130 g/kg did not cause any deaths or side effects in mice regarding body weight and biochemical parameters. This seems to confirm safety of Fuzi after long-time decoction. However, histopathological observations revealed an abnormal liver phenotype and a significant decrease of the liver index following FZ-120 treatment, indicating a potential hepatoxicity of FZ-120. By using a zebrafish model, we observed that FZ-120 at a dose range from 288 to 896 μg/ml caused considerable adverse events including arrhythmia, liver degeneration, yolk sac absorption delay, length decrease, and swim bladder loss, which clearly speak for acute toxicity on cardiovascular, digestive, development, and respiratory systems. The dose range of FZ-120 was lower than that used for clinical application in human beings. Moreover, UPLCMS revealed that FZ-120 still contained toxic aconitines that were not detectable by HPLC, which might explain its acute toxicity in zebrafish. We concluded that Fuzi is not sufficiently safe even after long-time decoction. The zebrafish model combined with UPLC-MS assay may represent an appropriate test system to unravel aconitinerelated acute toxicity.

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