Radioimmunoassay Method Research Articles

ПАТОЛОГИЯ МОЛОЧНЫХ ЖЕЛЁЗ ПРИ ХРОНИЧЕСКОМ ЭНДОМЕТРИТЕ У ЖЕНЩИН С РЕПРОДУКТИВНЫМИ НАРУШЕНИЯМИ

Background. The last decades are devoted to the search for various factors leading to the development of benign breast diseases, which are the risk of breast cancer. In gynecological diseases, the frequency of benign processes in the mammary glands increases to 76-97.8 %. Aims: this research is devoted to the study of the prevalence and structure of breast pathology in women with chronic endometritis and reproductive disorders. Materials and methods. The study included patients with reproductive disorders: 50 patients with histological verification of the diagnosis of chronic endometritis and 50 patients without chronic endometritis. Hormonal examination was carried out using radioimmunoassay and immunoenzymatic methods. Methods of mathematical statistics implemented Statistica 6.1 (USA) were used parametric criteria: Student's t-test and Fisher's F-criterion, the nature of the distribution was estimated using the methods of Kolmogorov - Smirnov with a significance level smaller and equal to 5 % (p < 0.05). To confirm the fact that the observed differences between the products of A x D and B x C are not random, we used the х2 criterion. Results and conclusions. In women with chronic endometritis, the pathology of the mammary glands is characterized by the presence of ectasia of the ducts of the mammary gland; clinically, it is manifested by lactorrhea. Benign dysplasia of mammary glands is accompanied by the presence of hyperestrogenia, hypoprogesteronemia, which is typical for ovarian dysfunction. It is possible to use a pathogenetically reasonable combination correction: anti-inflammatory therapy and the use of gestagens in women with reproductive disorders and chronic endometritis.

Optimization of reagent concentration for radioiodination of rat C-peptide II in development of radioimmunoassay procedure for rats

Rat C-peptide is a polypeptide molecule made up of 31 amino acids and secreted from pancreas into circulation in two isoforms I and II. Quantification of rat C-peptide II in rat serum is important as it is directly related to the diagnosis of carbohydrate metabolism abnormalities, pancreatic performance analysis, monitoring of hypoglycemia, and diabetes-related illness in rat model. The aim of the present work is to develop a tracer by chloramine-T method for radioimmunoassay (RIA) procedure and to determine the optimum amount of chloramine-T required for the preparation of stable radioiodinated product with a specific activity of around 24.97 MBq/μg, corresponding to 1 125I atom per molecule of the peptide. Tyrosylated rat C-peptide II was selected for the radioiodination procedure as rat C-peptide II does not contain either tyrosine or histidine which is mandatory for the incorporation of 125I atom to the rat C-peptide II. Tyrosylated rat C-peptide II was subjected to radioiodination by chloramine-T method with different concentrations of chloramine-T and sodium metabisulfite (MBS) to obtain a stable radiolabeled compound. Optimized reaction conditions relating to the concentration of chloramine-T (10 μg) and MBS (20 μg) yielded a stable 125I-rat C-peptide II with specific activity of 21.01 MBq/μg corresponding to 0.84 125I atoms per molecule of the peptide. Preparation of high integrity tracer of rat C-peptide II was achieved by combining one molecule of oxidant (chloramine-T) and two molecule of reductant (MBS).

Niveles de vitamina D en pacientes con lesión medular traumática en Chile

IntroductionVitamin D plays an important role in bone metabolism and in various extra-skeletal functions. The aim of this study was to determine the prevalence of vitamin D deficiency in patients with traumatic spinal cord injury (TSCI) and to describe its association with the characteristics of TSCI and patients’ demographic characteristics. Material and methodA prospective descriptive study was conducted in 76 patients treated at the Rehabilitation Department of Hospital del Trabajador between July 30, 2015 and July 30, 2016. Blood measurement of 25-hydroxyvitamin D (25-OH-D) was performed by the radioimmunoassay method and a structured survey. The results were analysed using descriptive statistics, the Kruskall-Wallis test, Wilcoxon signed-rank test, chi-square test, Fisher test and Spearman rank correlation coefficient. ResultsThe median age was 49.5 years (SD 15.05) and the time from TSCI was between 1 month and 36 years. The mean 25-OH-D level was 16.4ng/mL (4.2-45.9ng/mL). Vitamin D deficiency (<30 ng/mL) was found in 94.7% of the participants and severe deficiency (<10ng/mL) in 27.6%. No significant differences were found between the association of vitamin deficiency with sex, neurologic level, or severity of TSCI, season, amount of time of sun exposure or geographical area of residence. There was s an inverse correlation with a tendency to decrease 25-OH-D for each year from the onset of TSCI (Ro=-0.33, p value=.0040). ConclusionVitamin D deficiency is highly prevalent in patients with TSCI. An early diagnosis is important to provide adequate supplementation in order to decrease the prevalence of complications.

Central [CNS] and Peripheral [Gastric Tissue] Selective Monitoring of Somatostatin (SRIF) with Micro-Sensor and Voltammetry in Rats: Influence of Growth Factors (GH, EGF).

Somatostatin (SRIF) is widely distributed throughout the body, and regulates the endocrine system via interactions with various hormones, including the pituitary growth hormone, the thyroid stimulating hormone and the majority of the hormones of the gastrointestinal tract. SRIF is present in the central nervous system (CNS), where it affects rates of neurotransmission, and is also reported to be active in the intestinal tract, with evidence that stressed rats present a significant decrease in antral somatostatin-like immunoreactivity (SLI). Analysis of SRIF has mainly been carried out by means of radioimmunoassay methods. Here, we propose the use of an electrochemical method, such as voltammetry, applied with carbon-based sensors and, in particular, the combination of differential pulse voltammetry with treated carbon fiber micro electrodes (DPV-µCFE) to facilitate the analysis of such peptidergic electro active hormones in the rat striatum and gastric tissue; the effect of growth hormone (GH) and epidermal growth factor (EGF), in particular, upon the SRIF signal has been studied in such tissues.

Oral Supplementation of Parturient Mothers with Vitamin D and Its Effect on 25OHD Status of Exclusively Breastfed Infants at 6 Months of Age: A Double-Blind Randomized Placebo Controlled Trial.

Exclusively breastfed infants are at increased risk of vitamin D deficiency and many lactating mothers have been found deficient in 25OHD stores. To compare serum vitamin D levels in exclusively breastfed infants at 6 months of age with or without oral supplementation of 600,000 IU of vitamin D3 to mothers in early postpartum period. Exclusively breastfeeding term parturient mothers were randomized 24-48 hours following delivery to receive either 600,000 IU of vitamin D3 (Cholecalciferol) over 10 days in a dose of 60,000 IU/day or placebo. 25OHD levels were measured by Radio Immuno Assay method at recruitment and after 6 months in all mothers and their infants. Urinary calcium and creatinine ratio was measured to monitor adverse effects of vitamin D3 in both mothers and infants at 14 weeks and 6 months of age. X-ray of both wrists in anteroposterior view and serum alkaline phosphatase of infants were done in both groups at 6 months of age to look for evidence of rickets. Maternal profile was similar in intervention (A) and control (B) groups. Mothers' serum 25OHD levels at recruitment were also similar being 16.2 ± 9.3 ng/mL in group A and 14.1 ± 7.1 ng/mL in group B. After 6 months, 25OHD levels were 40.3 ± 21.6 and 22.9 ± 20.1 ng/mL in group A and group B mothers (p ≤ 0.00), respectively. The serum 25OHD levels in cord blood were 9.9 ± 5.7 and 8.9 ± 5.1 ng/mL, respectively, in infants born to mothers in intervention and control groups (p = 0.433). At 6 months of age, the serum 25OHD levels significantly (p < 0.00) raised to 29.1 ± 14.6 ng/mL in infants of group A compared to those of group B (15.7 ± 17.7 ng/mL). Four infants developed radiological rickets at 6 months of age, two infants each in intervention group and study group. As against 10 infants in the control group (16.94%), no infant in the study group had biochemical rickets. Urinary calcium and creatinine ratio in mothers and infants at 14 weeks and 6 months of age in both intervention and study group was within normal limits, indicating there was no adverse effects of oral administration of 600,000 IU of vitamin D3. Serum 25OHD levels of exclusively breastfed infants significantly rise at 6 months of age when their mothers are orally supplemented with 60,000 IU of vitamin D3 daily for 10 days in the early postpartum period in comparison to infants of vitamin D3 unsupplemented mothers.

Effect of Estrogen on Serum Total Cholesterol and Triglyceride Levels in postmenopausal women

Background: Hyperlipidemia is a major risk factor for cardiovascular diseases in postmenopausal women. Increased incidence of cardiovascular diseases in postmenopausal women may be due to hyperlipidemia caused by lower level of estrogen hormone.Objectives: The study was carried out to observe the association of serum estrogen with total cholesterol (TC) and triglyceride (TG) levels in postmenopausal women.Methods: This cross sectional study was conducted in the Department of Physiology, Dhaka Medical College, Dhaka, during the period of January to December 2011. A total of 90 females were selected from different areas of Dhaka city. Among them, 60 postmenopausal women with age ranging from 50 to 60 years were taken as study group and 30 apparently healthy premenopausal women with age ranging from 20 to 30 years were included as control group for comparison. The study parameters total cholesterol and triglyceride were estimated by enzymatic method in both groups. Serum estrogen level was estimated by Radioimmunoassay ( RIA) method in order to assess the hormonal level of both groups. Data was analyzed by Unpaired Student’s t- test and Pearson’s correlation co-efficient (r) test as applicable.Results: In this study, the mean serum TC level was higher in postmenopausal women than those of premenopausal women and result was statistically significant. The level of mean serum TG was significantly (p&lt;0.001) higher in postmenopausal women in comparison to those of premenopausal women. In postmenopausal women serum estrogen level was lower than premenopausal women and serum estrogen level showed negative correlation with TC level. .Again serum TG level also showed negative correlation with serum estrogen level. All these correlation were statistically non-significant.Conclusion: From the results of the present study it may be concluded that the serum total cholesterol and triglyceride levels are significantly higher in postmenopausal women that may be due to low level of estrogen.J Dhaka Medical College, Vol. 26, No.1, April, 2017, Page 25-31

PP.10.20] IMBALANCE BETWEEN T REGULATORY AND TH17 LYMPHOCYTE SUBSETS IN PATIENTS WITH RESISTANT HYPERTENSION AND PRIMARY ALDOSTERONISM

Objective: In the past several years, it has become evident that inflammation contributes to the elevation of blood pressure. Evidence from experimental models of hypertension and hypertensive patients suggests imbalance of T effector and regulatory subsets in hypertension. In our study we aimed to quantify blood T lymphocyte subsets in subjects with secondary hypertension- resistant hypertension (RHT) and primary aldosteronism (PA) characterized by increased BP levels, augmented cardiovascular risk in comparison to control subjects matched for age, sex and BMI (CG). Design and method: In an ongoing study we included 29 patients (20 M, 9F, mean age 54.9 ± 11.5) with RHT, 26 patients with PA (14 M,12F, mean age 54.8 ± 12.6) and 30 CG patients (15 M, 15F, mean age 50.4 ± 11.2). T cells characteristics from peripheral blood samples were studied by multicolour flow cytometry with intracellular staining for foxp3. The concentration of the angiotensin II in patients’ blood samples was determined by radioimmunoassay method. Data were analyzed using T test with Bonferroni correction for multiple comparisons. Results: Patients with RHT and PA were characterized by significantly higher BP values on ambulatory blood pressure monitoring as compared with CG. In comparison to CG, patients with RHT and PA had lower percentage of CD4+25+foxp3+ T cells (Tregs): consecutively RHT 5.5 ± 3.2%; PA 5.8 ± 4.3% vs CG 9.9 ± 10.6%; p = 0.01, p = 0.084. However, the percentage of CD3+CD4+IL-17+ T cells was higher in RHT 19 ± 20% and PA 12.9 ± 16.2%, vs CG 3.7 ± 2.4%; p < 0.01, p = 0.04. In patients with PA, in comparison to CG and RHT, lower angiotensin II concentration and higher aldosterone to renin ratio was observed. Conclusions: Our results indicate, that dysregulation of T cell activation with very significant increase of proinflammatory T cells and decrease in Tregs may play a role in the pathogenesis of RHT and PA, regardless of the renin-angiotensin-aldosterone activation.

How swimming affects plasma insulin and glucose concentration in Thoroughbreds: A pilot study

Low intensity exercise increases insulin-stimulated glucose uptake in skeletal muscle and decreases its plasma concentration. In this study, plasma insulin and glucose concentrations were evaluated 5min before and 5, 15, 25, 35, 45 and 60min after an IV bolus of glucose in 12 Thoroughbreds before and after 1 month of submaximal aquatraining exercise, monitored using heart rate and blood lactate. Plasma glucose concentrations were evaluated using a colorimetric enzymatic method, and plasma insulin concentrations with a solid-phase radioimmunoassay method. Pre-training plasma glucose concentrations at 15, 25 and 35min, area under the glucose curve and peak glucose concentration were significantly higher than post-training values (P<0.05). Baseline pre-training plasma insulin concentrations were significantly lower than in the post-training period, and plasma insulin was significantly higher at 45 and 60min in the pre-training period than the post-training period. These results indicate that aquatraining could improve insulin-glucose metabolism in horses.

Polymorphisms in Lep and Lepr Genes in Infants: Correlation with Serum Leptin Values in the First 6 Months of Life

ABSTRACTObjective: Because several studies indicate that polymorphisms in leptin (Lep) and leptin receptor (Lepr) genes play a central role in determining obesity, we analyzed 2 single nucleotide polymorphisms (SNPs) in the Lep gene (Lep G2548A and A19G) and one in the Lepr gene (Lepr A668G) to verify the effect of the 3 SNPs on leptin concentrations in infancy.Methods: We enrolled 80 healthy Caucasian infants under 6 months of age, who were genotyped for the 3 SNPs with amplification refractory mutation system–mismatch amplification mutation assay (ARMS-MAMA) real-time polymerase chain reaction (PCR). Serum leptin values were measured with a radioimmunoassay method. Statistical significance was set at p < 0.05.Results: There were no significant differences between individually analyzed leptin polymorphisms Lep G2548A and A19G and serum leptin levels (p > 0.05). Because we found that Lep G2548A and A19G are in linkage disequilibrium on chromosome 7, we performed the haplotype analysis for Lep G2548A and Lep A19G. We obtained higher serum leptin levels in infants with the GG/GG haplotype (p < 0.05). Regarding receptor, we found higher leptin levels in GG-genotype infants for Lepr A668G (p < 0.001). Considering the 3 SNPs together, we found higher serum leptin values in GG/GG-GG infants (LepG2548A/A19G-Lepr A668G; p < 0.001).Conclusion: We obtained higher serum leptin levels in infants with the GG genotype for Lepr A668G, with haplotype GG/GG for Lep G2548A/A19G, and with GG/GG-GG (LepG2548A/A19G-Lepr A668G); thus, it seems that the genotype GG could be a protector against obesity development in infancy and adulthood. Moreover, these data confirm that not variations in the Lep gene as well as in the Lepr gene could play a role in weight gain. Further studies are needed to evaluate the role of genetics and the environment in a predisposition toward obesity later in life.

A simple and fast LC-MS/MS method with a very high sensitivity for the measurement of raltitrexed in human plasma.

Raltitrexed is a thymidylate synthase inhibitor that can be administered safely to patients with cardiovascular disease or dihydropyrimidine dehydrogenase deficiency, as opposed to 5FU. The recommended dose of 3mg/m2 every 3 weeks often leads to toxicity. Interestingly, the 2mg/m2 every 2 weeks dose appears to be less toxic. A pharmacokinetic trial was then performed by our team to investigate such phenomenon. However, there are currently, two main methods for RTX measurement described in the literature: a radioimmunoassay (RIA) and chromatographic-based methods with either UV or mass spectrometry detections. The RIA methods: display a low limit of quantification (below 1μg/L), but also a low extent of linearity for the calibration curve. The chromatographic-based methods: include high level of calibrators, but have poor sensitivity (>2μg/mL). If a high sensitivity is essential to satisfactorily describe the elimination of RTX, high concentrations in the calibration curve are also needed to avoid bias linked to the dilutions of the samples. A new LC-MS/MS method was then developed that allows to simultaneously measure very low (0.1μg/L) and very high (3000μg/L) concentrations in the same run. Moreover, the extraction steps are very simple and fast with mainly a precipitation and a filtration steps. This method was validated following the EMA recommendations. In view of the extent of the calibration curve, the carry-over effect was more deeply investigated. With this method, it was possible to measure RTX in samples taken 3 weeks after the administration. Taken together, this method allows to simply and quickly measure RTX in plasma of patients.

Fecal cortisol radioimmunoassay to monitor adrenal gland activity in the bottlenose dolphin (Tursiops truncatus) under human care

AbstractFecal glucocorticoid measurement is an important noninvasive tool to monitor animal health. A radioimmunoassay (RIA) method was developed to measure fecal cortisol in bottlenose dolphins under human care. The method was used to measure baseline hormone levels and evaluate the adrenal response to environmental challenges in a small number of individual dolphins. The method was validated by precision and accuracy tests and by comparison with liquid chromatography‐mass spectrometry (LC‐MS). The parallelism test suggested few matrix interferences. The assay showed a good degree of precision within assay (CV = 5.4%) and between assays (CV = 4.1%). The RIA significantly correlated with the LC‐MS method (r = 0.838, P &lt; 0.01). The recovery test and the comparison between RIA and LC‐MS suggested that the RIA slightly underestimates fecal cortisol concentrations, although the degree of accuracy was good. This study established that bottlenose dolphins excrete appreciable amounts of fecal cortisol (healthy subjects: 0.2–9.5 ng/g). Therefore, chronic HPA axis activation may be monitored in fecal samples by immunoassays after validating a suitable extraction protocol. The RIA could discriminate conditions of stimulation (pregnancy, parturition, isolation, transportation) and inhibition (diazepam administration) of the HPA axis and may, therefore, be useful for monitoring dolphin well‐being.

Studies on the Relationship Between Chromium(III) ion and Thyroid Peroxidase Activity in Sera of Patients with Thyroid Dysfunction

The objective of this study was to determine the concentration of trace element chromium(III) and thyroid peroxidase activity in human serum , and to find a relationship between the concentration of chromium(III) and thyroid peroxidase activity in serum of patients with hypothyroidism, hyperthyroidism and healthy subjects. Serum thyroid peroxidase was measured by enzyme linked radioimmunoassay(ELISA) method and chromium determination was by atomic absorption spectrophotometer .Comparing the values of chromium concentration and thyroid peroxidase activity in both samples showed that there were significant positive correlations between chromium levels and thyroid peroxidase activity(P&lt;0.01, r=0.11). The results showed that Serum thyroid peroxidase activity and chromium levels were significantly higher in hyperthyroidism patients (152.0±4.6 IU/L) (2.159±0.15 ppb) respectively than normal (51.0±1.8 IU/L) (0.378±0.024 ppb) respectively and lower than normal in hypothyroidism patients (35.0±0.31 IU/L)(0.099±0.011 ppb) respectively.

The Involvement of Corin in the Progression of Diabetic Erectile Dysfunction in a Rat Model by Down-Regulating ANP /NO/cGMP Signal Pathway.

This study was aimed to analyze the potential role of Corin in the procession of diabetic ED and to explore the underlying mechanism. Diabetic ED rat model was constructed and the characteristics of diabetic ED and control rats were recorded at 4, 8, 12, and 16 weeks. qRT-PCR and Western bloting were used to detected the mRNA and protein levels. Intracellular cGMP detection was accomplished using a commercial radioimmunoassay method. Vascular endothelial cell from rat corpus cavernosum spiral artery was isolated and transfected with si- Corin to analyzed the potential role of Corin. Cell viability was assessed using crystal violet. The results showed that diabetic ED rats showed significantly higher glucose level, and lower body weight, ICP level, and ICP/MAP ratio at 12 and 16 weeks in diabetic ED rats compared with control rats. The protein levels of Corin, atrial natriuretic peptide (ANP) and eNOS, and the level of cGMP were significantly down-regulated in corpus cavernosum in diabetic ED rats, revealing the potential role of Corin in NO-associated diabetic ED. Further, studies proved that defect of Corin not only inhibited the vascular endothelial cell viability in high-glucose condition, but also suppressed ANP, eNOS, and cGMP expression in vascular endothelial cells. To sum up, Corin contributes to the progression of diabetic ED and the underlying mechanism is associated with the down-regulation of ANP /NO/cGMP signal pathway. J. Cell. Biochem. 118: 2325-2332, 2017. © 2017 Wiley Periodicals, Inc.

Evaluation of a commercially available radioimmunoassay and enzyme immunoassay for the analysis of progesterone and estradiol and the comparison of two extraction efficiency methods

The measurement of progesterone (P4) and estradiol (E2) is essential for monitoring reproductive cycles and can aid in diagnosing the cause of poor reproductive performance in dairy cattle. Readily available, reproducible, accurate, non-radioactive assays are needed for the assessment of P4 and E2 in bovine serum. The gold standard for hormone assessment, radioimmunoassay (RIA), was compared with enzyme-linked immunoassay (EIA). Serum collected from various points in the estrous cycle was extracted with radiolabeled P4 (ie, 3H-P4; HE) and without 3H-P4 (CE) before being used in the assay. For the assessment of P4, there is a great degree of correlation between the RIA and EIA (adjusted R-square = 0.95; Pearson correlation coefficient (PCC) = 0.98, P < 0.001). A difference between the RIA and EIA methods was not detected for E2 concentrations (P = 0.16), but the correlation between techniques was poor (adjusted R-squared = 0.73; PCC = 0.87, P = 0.002). There was no difference in the serum extraction efficiency as measured with 3H-P4 as opposed to without (P = 0.94). The two methods for the measurement of serum extraction efficiency were highly correlated (adjusted R-square = 0.83; PCC = 0.92, P < 0.001). The concordance correlation coefficient (CCC) showed an excellent agreement between RIA and EIA for P4 determination (0.89) and between HE and CE methods (0.90). Although the 95% limits of agreement of the Bland-Altman plots encompassed 89% (8/9) and 92% (12/13) of the differences between methods for P4 quantification and extraction respectively, the CCC indicated an excellent agreement among them. The CCC between RIA and EIA for E2 quantification was 0.68 which corresponds with a fair agreement; however, the 95% limits of agreement of the Bland-Altman plot encompassed 100% (9/9) of differences between methods. The EIA and CE methods are comparable alternatives to the RIA and HE methods, respectively and can be used to quantify P4 and E2 for bovine serum.

Kisspeptin Increases Luteinizing Hormone Secretion in Food Deprived D-Lys3-GHRP6-Treated Rats

Background: Kisspeptin stimulatesthe reproductive axis while food deprivation or ghrelin inhibits it. The aim of the study was to determine the effects of third cerebral ventricle injection of kisspeptin on mean plasma luteinizing hormone (LH) levels in food deprived D-Lys3-GHRP6-treated rats. Materials and Methods: In this experimental study, five fed rats received third cerebral ventricular injection of salineat 09:00- 09:30 h. Also, twenty food deprived rats in four groups (n=5 in each group) received third cerebral ventricular injection of saline, kisspeptin(1nmol), D-Lys3–GHRP-6 (20nmol) or D-Lys3-GHRP-6 (20nmol) and kisspeptin(1nmol). Blood samples were collected via the tail vein. Plasma LH concentration was measuredby using LH kit and the method of radioimmunoassay (RIA). Results: Mean plasma LH level in food-deprived rats decreased significantly compared to fed rats. Kisspeptin significantly increased the mean plasma LH concentrationcompared to fed or food deprived saline group. Injection of kisspeptin in D-Lys3 -GHRP-6-pre-treated rats significantly increased the mean plasma LH concentration compared to fed saline, food deprived saline, alone kisspeptin or alone D-Lys3 -GHRP-6 groups. Conclusion: Decrease of ghrelin pathway activity may partly be involved in the mediating the stimulatory effects of kisspeptin on hypothalamic-pituitary-gonadal axis. [GMJ. 2017;6(1):39-43]

Analgesia effect of baicalein against NTG-induced migraine in rats

BackgroundMigraine is a complex nervous system disease characterized by typical throbbing and unilateral headache, which causes severe healthy and social issues worldwide. The purpose of this study was to investigate the effect of baicalein (BAI) on the treatment of migraine. Material and methodsTwenty-four rats were randomly divided equally into four groups, including a blank group, model group, positive group (ibuprofen tablets 82mg/kg), and BAI group (60mg/kg). All rats were intragastrically treated with the corresponding treatment for 10 consecutive days, and they were subcutaneously injected with NTG (10mg/kg) 1h after the last treatment, except in the blank group. After model establishment, the behaviors of all rats, including scratching head and shaking body were observed continuously for 100min. Four hours after NTG treatment, all rats were anaesthetized and the blood was collected. Thereafter, nitric oxide (NO) in plasma was determined by colorimetric method, the level of calcitonin gene-related peptide (CGRP) and endothelin (ET) were detected by radioimmunoassay method. In addition, immunohistochemistry was applied to detect c-Fos neuronal activity in trigeminal nucleus caudalis (TNC). ResultsBehavioral research showed that BAI administration alleviated the hyperalgesia in migraine rats. Compared with the model group, the levels of NO and CGRP in BAI administration groups were markedly decreased (p<0.01), and the levels of ET was significantly increased (p<0.01). Meanwhile, immunohistochemistry results showed that NTG treatment significantly activated c-Fos neurons while BAI treatment inhibited the expression of c-Fos. ConclusionsBAI could alleviate the migraine-like headache induced by NTG, which is related to the regulation of vasoactive substances. These findings may contribute to the further study of BAI as a potential drug for migraine pharmacotherapy.

Métodos para cuantificar la testosterona sérica en pacientes con cáncer de próstata sometidos a castración: una revisión sistemática

BackgroundThe clinical practice guidelines recommend measuring serum testosterone in patients with prostate cancer (PC) who undergo castration. The serum testosterone concentration should be <50ng/dL, a level established by using a radioimmunoassay method. The use of chemiluminescent immunoassays (IA) has become widespread, although their metrological characteristics do not seem appropriate for quantifying low testosterone concentrations. The objective of this review is to analyse the methods for quantifying testosterone and to establish whether there is scientific evidence that justifies measuring it in patients with PC who undergo castration, through liquid chromatography attached to a mass spectrometry in tandem (LC-MSMS). Material and methodsWe performed a search in PubMed with the following MeSH terms: measurement, testosterone, androgen suppression and prostate cancer. We selected 12 studies that compared the metrological characteristics of various methods for quantifying serum testosterone compared with MS detection methods. ResultsIAs are standard tools for measuring testosterone levels; however, there is evidence that IAs lack accuracy and precision for quantifying low concentrations. Most chemiluminescent IAs overestimate their concentration, especially below 100ng/dL. The procedures that use LC-MSMS have an adequate lower quantification limit and proper accuracy and precision. We found no specific evidence in patients with PC who underwent castration. ConclusionsLC-MSMS is the appropriate method for quantifying low serum testosterone concentrations. We need to define the level of castration with this method and the optimal level related to better progression of the disease.

Effects of Anethum graveolens L. (dill) Seed and Leaf Aqueous Extracts on the Growth of Mammary Gland Alveolar Buds of Rat

The effects of dill seed aqueous extract on the sex hormones have already been reported in our previous studies. In this study, we examined the oral administration of dill leaf and seed extracts on the mammary gland tissue that were exposed to these hormones. Adult female rats were divided into 3 groups of control, dill leaf and seed aqueous extracts (0.5 g/kg) treated groups (n = 7). After 25 days, the rats that were in estrus phase were dissected, and their pelvic mammary glands were spread on slides and stained by carmine. Then, the alveolar buds 1 and 2 (AB1 and AB2) and lobules of the whole mount mammary glands were counted and scored in tip, central and distal parts of mammary gland, then the ratio of mammary gland differentiation 1 and 2 (Dif1 and Dif2) was calculated. Estradiol concentration was measured via radioimmunoassay method and Progesterone, FSH and LH concentrations were measured using immunoradiometric assay. The data were analyzed by One-Way ANOVA test. The results showed that AB1 decreased but AB2 increased in total mammary gland of the experimental groups; however, these changes were not statistically significant. The numbers of lobules and Dif2 in the mammary gland were significantly increased in dill leaf and seed extracts treated groups. Progesterone concentration increased in dill seed extract-treated group. The results indicated that oral administration of dill seed and leaf aqueous extracts increases the mammary glands growth; therefore, dill may be considered as lactiferous herb.

Vitamin D deficiency and its impact on asthma severity in asthmatic children.

BackgroundDespite obtaining evidences on association between vitamin D and development of lung in fetus, little is known about vitamin D level and its impact on severity of asthma in children. The present study aimed to assess the relationship between the asthma severity and vitamin D deficiency in asthmatic children.MethodsThis case-control study was conducted on 106 individuals including asthmatic (n = 53) and healthy children (n = 53) who referred to Mofid hospital in Tehran in 2013. The level of serum vitamin D in both groups was measured by radioimmunoassay method at the reference lab and was categorized as sufficient (> 30 ng/ml), insufficient (20 to 30 ng/ml), or deficient (< 20 ng/ml). The control status of asthma in patients group was classified as controlled, partially controlled, and uncontrolled.ResultsIn the groups with and without asthma, the prevalence of vitamin D deficiency was 73.6 and 49.1%, and the prevalence of vitamin D insufficiency was 18.9 and 18.9%, while normal vitamin D level was revealed in 7.5 and 32.1%, respectively with a significant difference (p = 0.005). Using the multivariate logistic regression analysis, the presence of asthma was associated with reduced level of vitamin D (OR = 1.068, 95% CI: 1.027–1.110, P = 0.001). In this context, the risk for asthma in the children with vitamin D deficiency was 6.3 times of those with normal vitamin D level. Although the presence of asthma was strongly associated with reduced level of vitamin D in serum, neither severity of asthma nor control status of asthma were associated with vitamin D deficiency.ConclusionThe presence of vitamin D deficiency effectively predict increased risk for childhood asthma; however the severity or control status of this event may not be predicted by confirming vitamin D deficiency.

Pharmacodynamic action and mechanism of Du Liang soft capsule, a traditional Chinese medicine capsule, on treating nitroglycerin-induced migraine

Ethnopharmacological relevanceDu Liang soft capsule (DL) is a traditional Chinese medicine for treating migraines; it is made from two Chinese herbs, including LigusticumstriatumDC., root; Angelica dahurica (Hoffm.) Benth. & Hook.f. ex Franch. & Sav., root. Aim of the studyIn the present study, we aimed to elucidate the pharmacodynamic action of DL and its mechanism in an animal model of migraines induced by glyceryl trinitrate (GTN). Materials and methodsSixty rats were randomly divided into six groups, including a normal control group, model control group, positive group (Sumatriptan 0.006gkg−1), and three DL groups (0.44, 1.31 and 3.93gkg−1). All rats were intragastrically treated with the corresponding treatment for 7 consecutive days, and they were subcutaneously injected with GTN (10mgkg−1) 30min after the last treatment, except in the normal control group. After model establishment, the behaviors of all rats, including head scratching, cage climbing, and the development of red ears were observed continuously by digital camera every 30min for 3h. Four hours after GTN treatment, all rats were anaesthetized and the blood and tissue samples were collected. Plasma calcitonin gene related to peptide (CGRP) and endothelin (ET) levels were measured using the radioimmunoassay method, and serum NO was determined by the colorimetric method. Afterwards, the brainstem tissues were dissected and washed with physiological saline, and divided evenly into two parts. One part was used to test the monoamine levels, including levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA), by the fluorometric method, and the other part was used to determine the nuclear factor kappaB (NF-κB) p65, nuclear c-fos, inducible nitric oxide synthase (iNOS), interleukin (IL)-1β (IL-1β), and cyclooxygenase-2 (COX-2) levels by Western blot analysis. ResultsIn the pharmacodynamic action assay, DL (1.31 and 3.93gkg−1) greatly improved the abnormal behaviors of migraine rats, including head scratching and cage climbing, and the development of red ears. In the mechanism assay, compared with the control group, the plasma CGRP and serum NO levels and the brainstem 5-HT, NE and DA levels in the DL administration groups were significantly decreased; and the plasma ET levels were remarkably increased. Moreover, down-regulation of NF-κB p65, c-fos and pro-inflammatory cytokines, including iNOS, IL-1β and COX-2 in the brainstem in the DL administration groups were observed by Western blot analysis. ConclusionsThe above results suggested that DL has a therapeutic effect on migraines, and its mechanism may be related to adjusting the level of neurotransmitters and vasoactive substances, consequently relieving neurogenic inflammation.