Determination Of Hormones Research Articles

Quantitative determination of plant hormones and derivatives in biogas slurry using on-line solid phase extraction with high performance liquid chromatography (On-Line SPE HPLC)

A novel method using on-line solid phase extraction coupled with high performance liquid chromatography (On-line SPE HPLC) was developed to measure three important plant hormones and two indole derivatives in the biogas slurry. These measured substances include abscisic acid (ABA), gibberellins (GA), indoleacetic acid (IAA), tryptophan (TRP) and skatole (IMD), respectively. This method could continuously perform extraction of ABA, GA, IAA, TRP and IMD from different concentrations of biogas slurry samples without complicated pretreatment avoiding lost, and then the extracts were analyzed by high performance liquid chromatography (HPLC). After optimized the analytical condition, a baseline separation of three plant hormones and two indole derivatives was accomplished within 35 mins. The results showed excellent linearity (R2 values of 0.9930-0.9999) and intra- and inter-day precisions (in the range of 3.8-7.2% and 3.9-7.5%), respectively. The limits of detection and quantification were in the range of 0.16-0.47 μg·mL-1 and 0.53-1.57 μg·mL-1, respectively. The determination performance was satisfactory with the proposed method. The results showed that the developed method was suitable for analysis of trace plant hormones in the biogas slurry.

Quantitative determination of plant hormones and derivatives in biogas slurry using on-line solid phase extraction with high performance liquid chromatography (On-Line SPE HPLC)

A novel method using on-line solid phase extraction coupled with high performance liquid chromatography (On-line SPE HPLC) was developed to measure three important plant hormones and two indole derivatives in the biogas slurry. These measured substances include abscisic acid (ABA), gibberellins (GA), indoleacetic acid (IAA), tryptophan (TRP) and skatole (IMD), respectively. This method could continuously perform extraction of ABA, GA, IAA, TRP and IMD from different concentrations of biogas slurry samples without complicated pretreatment avoiding lost, and then the extracts were analyzed by high performance liquid chromatography (HPLC). After optimized the analytical condition, a baseline separation of three plant hormones and two indole derivatives was accomplished within 35 mins. The results showed excellent linearity (R2 values of 0.9930-0.9999) and intra- and inter-day precisions (in the range of 3.8-7.2% and 3.9-7.5%), respectively. The limits of detection and quantification were in the range of 0.16-0.47 μg·mL-1 and 0.53-1.57 μg·mL-1, respectively. The determination performance was satisfactory with the proposed method. The results showed that the developed method was suitable for analysis of trace plant hormones in the biogas slurry.

GC-MS/MS Determination of Steroid Hormones in Urine Using Solid-Phase Derivatization as an Alternative to Conventional Methods

Solid-phase analytical derivatization (SPAD) is a promising hybrid sample preparation technique combining the clean-up and preconcentration of the sample in a single step. In this work, a novel SPAD method based on the preparation of trimethylsilyl (TMS) derivatives of steroid hormones (testosterone, estrone, DHT, estriol, estradiol, and progesterone) in Phenomenex Strata C18-E (100 mg, 1 mL) cartridges has been developed and applied for their GC-MS/MS determination in human urine samples. The proposed procedure allows the detection and quantification of steroids with limits of 1.0–2.5 and 2.5–5 ng/mL, respectively. These characteristics are comparable with those obtained with a conventional liquid–liquid extraction, while the recovery of analytes in the proposed SPAD procedure is higher. The major advantages of SPAD are a short derivatization time, high efficiency, and the possibility to automatize the procedure. However, its cost-effectiveness in routine practice is still questionable.

Hypothyroidism Induced Hyponatremia: A Case Report.

Thyroid hormones have various effects on the body which include electrolyte and water hemostasis. It is also involved in renal development and physiology. Hyponatremia is a serious electrolyte imbalance that can be associated with the involvement of different body systems and a wide range of deleterious changes. We report a case of a 62-years-old female with the symptoms of severe hyponatremia like altered mental sensorium with a serum sodium level of 110 meq/l. After ruling out other causes, a final diagnosis of hypothyroidism was made. On treating hypothyroidism the symptoms of hyponatremia were resolved. Therefore, thyroid stimulating hormone determination is mandatory during the evaluation of hyponatremia. And, those patients should be treated with fluid restriction and treatment of hypothyroidism.

Simultaneous Determination of Antibiotics, Mycotoxins, and Hormones in Milk by an 8-17 DNAzyme-Based Enzyme-Linked Immunosorbent Assay.

The simultaneous detection of three kinds of small-molecule contaminants (antibiotics, mycotoxins, and hormones) in milk was realized by using an 8-17 DNAzyme-based fluorescent enzyme-linked immunosorbent assay (ELISA), in which 8-17 DNAzyme was utilized as the catalytic enzyme for amplifying the signal. Compared with the conventional ELISA in which horseradish peroxidase is used as the catalyzing factor, this 8-17 DNAzyme-based ELISA could achieve multicolor signal output with lower detection limits. The linearities for chloramphenicol, 17β-estradiol, and aflatoxin M1 were in the range of 0.3 ng/mL-3 μg/mL, 3 ng/mL-3 μg/mL, and 3 pg/mL-3 ng/mL with quantitation limits of 0.3, 3, and 0.003 ng/mL, respectively. This proposed scheme demonstrated that the 8-17 DNAzyme might be an effective substitute for horseradish peroxidase in ELISA for the development of ultrasensitive and multicolor fluorescence immunoassay, which would stimulate the development of ELISA in a new orientation.

Experimental validation and molecular docking to explore the active components of cannabis in testicular function and sperm quality modulations in rats

BackgroundData available support that ninety percent of male infertility cases are due to low sperm counts. There is a scarcity of data on the medicinal effects of cannabis on fertility. This study evaluated testicular function and sperm quality modulation with cannabis in rats.MethodologyTwenty-five male Wistar rats were randomly grouped into five: A, B, C, and D, each group have 5 rats. A (control): 0.2 ml 2% DMSO, B (vitamin C): 90 mg/kg body weight, C, D, and E were administered: 5 mg/kg, 10 mg/kg and 20 mg/kg body weight of ethanolic leaf extract of cannabis (ELEC) respectively. The rats were sacrificed 24 h after the last day of the 60 day oral administrations. Flavonoids were the predominant phytochemical present in the extract while quercetin, kemferol, silyman and gallic acid were identified.ResultsThe results showed a significant improvement (p < 0.05) in sperm quality and a significant increase in the concentrations of follicle-stimulating hormone, luteinizing hormone, triglycerides, cholesterol, and total protein determination compared to the normal control. Similarly, there was a significant increase (p < 0.05) in the activities of acid phosphatase, alkaline phosphatase, and superoxide dismutase compared to the normal control. RAC-alpha serine/threonine-protein kinase (AKT1)-silymarin complexes (-8.30 kcal/mol) and androgen receptor (AR)-quercetin complexes (9.20 kcal/mol) had the highest affinity.ConclusionThe antioxidant effects of the flavonoids in the ethanolic extract of cannabis may have protected testicular and sperm cells from oxidative damage. Biochemical processes and histopathological morphology were preserved by cannabis. The docking prediction suggests that the bioactive principle of cannabis may activate the androgenic receptors. The androgenic receptor modulation may be attributed to silymarin and quercetin.

Expression of Hormones, Cytokines, and Antioxidants in Heat-Stressed Subfertile Female Dromedaries.

Simple SummaryHeat stress imposes a high burden on domestic animals’ productive and reproductive performance. Due to the long hot summer, drought, and shortage of green fodders, camels raised in the desert suffer a lot of reproductive inefficiencies. This animal represents one of the main wealth sources for the desert inhabitants. Several fertility disorders have been discovered, leading to frequent breeding without pregnancy. This study aimed at exploring blood metabolites such as metabolic and reproductive hormones, cytokines, and antioxidants to be monitored as bio-indictors for subfertility in female camels. The results confirmed that none of the tested metabolic hormones and glucose revealed differences among fertile and subfertile females. However, FSH, inhibin, IL-ß, nitrous oxide, and glutathione revealed remarkable differences between fertile and subfertile females, which would be reliable tools to predict subfertility statuses in this animal. IL-ß revealed higher levels in the cases with genital inflammations. The normal profiles in control females revealed the highest FSH, and the lowest inhibin were vice versa in all subfertile females. Nonetheless, nitrous oxide and glutathione would also be reliable bio-indicators for judging the fertility status.The prevailing hot climate imposes heavy burdens on the productivity of the camel, goat, and sheep herds raised in the Gulf desert. Due to the lack of a reliable indicator for the various subfertility statuses in camel females, this study aimed to investigate the expression of inhibin, TGFά, ILß, FSH, sex and metabolic hormones, and antioxidants for the fertility status in camel females. Eighty-two subfertile and five fertile females were admitted to the university clinic with the complaint of repeat breeding with failed conception. The animal’s genital tracts were examined for reproductive soundness. Blood samples were withdrawn for hormonal, cytokines, and antioxidants determinations. Subfertile females were categorized into six groups; endometritis (EN, 28), inactive ovaries (IO, 20), ovarian hydrobursitis (BU, 19), vaginal adhesions (VA, 7), salpingitis (SA, 4), and cervicitis (CE, 4). Results revealed a significant increase in inhibin in all groups compared to control (68.2, 66.4, 61.8. 58.8, 58.3, 55.8, and 36 pg/mL, in CE, VA, IO, BU, EN, SA, and CON, respectively). TGFά, dehydroepiandrosterone (DHEA), and progesterone were not different among groups, whereas IL-ß differed among groups. FSH, estradiol, nitrous oxide, and glutathione were higher in CON compared with other groups. In conclusion, reproductive failures in camel females are reflected in the imbalances of endocrine, cytokines, and antioxidants bio-indicators.

Hormonal and genetic causes of poor response to controlled ovarian stimulation in women of late reproductive age

Objectives: to investigate the hormonal and genetic determinants of the poor response to controlled ovarian stimulation (COS) in women of late reproductive age.Materials and methods. 130 patients from assisted reproductive technology programs were examined. The main group of 80 older reproductive aged patients (35 years and older) was divided into 2 subgroups according to the POSEIDON criteria. Subgroup 1 included 34 women with a predicted poor response to COS, subgroup 2 – 46 patients with a predicted normal response to COS. The comparison group consisted of 50 patients under the age of 35 with a predicted normal response to COS.The pituitary hormones (luteinizing, follicle-stimulating hormones, prolactin), steroid hormones (testosterone, estradiol, progesterone), cortisol, anti-Mullerian hormone (AMH) and inhibin B values was determined in blood serum by the ELISA. Blood was taken on the third day of the menstrual cycle. A molecular genetic study of FSHR gene polymorphism (Ala307Thr, Ser680Asn) was performed.Results. AMH decrease, estradiol increase and inhibin B decrease in potentially poor responders to COS may be an additional indication of reduced ovarian reserve. The level of follicle-stimulating hormone increases with age, but does not have high specificity as a marker of ovarian reserve. Elevated levels of cortisol emphasize its role in reproduction and correspond to increased stress value. The effectiveness of IVF if there were 4 or more mature oocytes obtained is associated with higher levels of AMH and inhibin B, and lower estradiol and cortisol values. The multifaceted dependence of the COC effectiveness in women of older reproductive age on the parameters of the hormonal profile, confirmed by correlation analysis, reflects the complexity of the reproductive function implementation with the use of auxiliary reproductive technologies in such patients. The combination of AA/SS genotypes for the Thr307Ala and Asn680Ser polymorphisms of the FSHR gene can be an additional marker of COC inefficiency.Conclusions. Determination of the hormones levels (inhibin B, estradiol and cortisol), the study of Thr307Ala and Asn680Ser polymorphisms of the FSHR gene can provide additional information for predicting the response to COS in women of older reproductive age.

Determination of hormones in urine by hollow fiber microporous membrane liquid–liquid extraction associated with 96-well plate system and HPLC-FLD detection

In this work, hollow-fiber microporous membrane liquid–liquid extraction (HF-MMLLE) was associated with a 96-well plate system for the determination of estrone, 17-β-estradiol, estriol and 17-α-ethinylestradiol in urine samples. This method exhibited some advantages, such as low cost, easy application, high-throughput and environmentally-friendly aspects. The type of organic solvent to fill the membrane, ionic strength effect, sample dilution, extraction and desorption time, and desorption solvent were examined. After the optimizations, the conditions were comprised of 45 min of extraction, 1-octanol as organic solvent and 15% (w/v) of NaCl; methanol was used as desorption solvent, and the desorption time was fixed at 10 min. The dilution of the sample increased the sensitivity due to the reduction of matrix effects; thus, urine samples were diluted 40-fold. The limits of detection ranged from 0.03 μg L−1 for 17-β-estradiol to 15 μg L−1 for estrone, and the limits of quantification ranged from 0.1 μg L−1 for 17-β-estradiol to 10 μg L−1 for estrone. The intra-day precision varied from 1.0% for estriol to 13.3% for 17-α-ethinylestradiol, and inter-day precision varied from 7.3% for estrone to 18.1% for estriol. The relative recoveries varied from 82 to 118%.

Associations between ubiquitin, follicle-stimulating hormone, and sex steroid hormones in the failed to conceive female dromedary camels raised in hot climates.

Background and Aim:The reproductive management of female dromedary camels involves traditional implications that are widespread among desert camel raisers. Several subfertility clinical manifestations impede pregnancy and elongate the interval between parturitions. Ubiquitin is a novel-specific protein, referred to recently as a biomarker for reproductive performance in male and female mammals. Therefore, this study aimed to investigate the association between subfertility clinical status and the peripheral levels of ubiquitin versus follicle-stimulating hormone (FSH), progesterone, and estradiol.Materials and Methods:According to the clinical diagnoses, 80 female dromedaries admitted to the university clinic were categorized into six female groups suffering from endometritis (EN, 28; 35%), inactive ovaries (IO, 18; 22.5%), ovarian hydrobursitis (BU, 19; 23.75%), vaginal adhesions (VA, 7; 8.75%), salpingitis (SA, 4; 5%), and cervicitis (CE, 4; 5%). In addition, five normal fertile non-pregnant females served as controls (CONs). All animals underwent ultrasonography and blood sampling for hormone and ubiquitin determinations.Results:The results revealed a significant (p < 0.05) increase in ubiquitin in the CE (577.22 pg/mL) and VA (670.92 pg/mL) females. However, lower ubiquitin levels but still higher than the CON were noted in females with other symptoms (225.76, 425.79, 394.02, 414.96, and 393.92 pg/mL in the CON, BU, SA, IO, and EN, respectively). Concomitantly, the mean levels of FSH revealed a similar trend, showing higher (p < 0.05) levels in CE (2.79 mIU/mL) and VA (2.5 pg/mL) females. In contrast, no change was observed in FSH among other groups than CON (2.11, 2.17, 2.01, 2.24, and 2.13 mIU/mL in CON, BU, SA, IO, and EN, respectively). There was no difference in the progesterone levels among groups; however, estradiol-17ß levels significantly differed (p < 0.01), showing the highest level (629.15 pg/mL) in the SA group with no significant difference among other groups.Conclusion:Thus, ubiquitin could be used as a biomarker for genital tract inflammation in female camels raised in hot climates.

МІКРОКЛОНАЛЬНЕ РОЗМНОЖЕННЯ ФУНДУКА

For the rapid introduction of large quantities of high-quality hazelnut planting material, it is important to use various modifications of microclonal propagation: classic on gel media, bioreactors with periodic flooding (TiS) and photoautotrophic methods. At the first stage of classical methods there are three main problems: phenol formation; endogenous contamination; selection of trophic and hormonal determinants of in vitro ontogenesis. In cases of infection of mother plants with viruses, viroids, it is mandatory to use meristem explants with a size of not more than 0.3 mm, followed by diagnosis of the effectiveness of recovery. To prevent self-intoxication with phenol-like oxidation products, a set of measures is used to prepare explant donors and modify nutrient media. The most common artificial nutrient media are Driver and Buzzard (DKW) and Nas and Read (NRM). Hazelnuts are sensitive to excess nitrogen and calcium and copper deficiency in artificial nutrient media. On media with a high N content, regenerants have shortened and thickened shoots, often with signs of hypehydration. Inhibition of calcium uptake by nitrogen leads to necrotization of shoot tips and root tips. The incorporation of iron ions into the metabolism of a plant depends on their valence and the form of chelating agents. During the multiplication and induction of rhizogenesis in aseptic conditions, mainly synthetic cytokinin benzylaminopurine and auxin indolylbutyric acid are used. The effectiveness of hormones increases with the use of combinations within groups, in particular cytokinins are benzylaminopurine and kinetin. At the stage of multiplication there is a predominance of cytokinins over auxins, and at the stage of rhizogenesis the number of auxins in the environment is greater than the number of cytokinins. Of the organic components, vitamins B1, B6, C, PP, amino acids and inositol are added to the environment. Ex vitro acclimatization of plants is carried out on peat-pearlite substrates in closed soil or modules of photoautrophic microclonal propagation with increased light intensity and high carbon dioxide content. The photoautotrophic method combines animation, rhizogenesis and acclimatization at the same time. The effectiveness of adaptation to factor-nestatic conditions increases with mycorrhizae by fungi of the genera Glomus, Trichoderma, Tuber in the transfer of plants from aseptic conditions.

Analysis of Thyroid Functional Status of the Covid-19 Survivors: A Single Center Study

Background: Numerous researches had illustrated the long-term impact of COVID-19 on thyroid gland worldwide. The aim of this study is to observe similar consequences among Bangladeshi population since there is not much information available. Materials and Methods: A total of 58 adult patients with a mean age of 29.53±9.005 years were studied to observe the long-term effect ( &gt; 6 months to 2 years) of COVID-19 infection, on their thyroid functions. Through convenience sampling, COVID-19 positive patients were selected, excluding any previously known thyroid illness or surgery, other endocrine complaints, pregnancy, COPD or bronchial asthma, hypertension, malignancy, or kidney disease. Thyroid functional status was assessed by quantitative determination of thyroid hormones and autoantibodies. Results: Between a case group of 58 adult COVID-19 survivors and a control group of 27 healthy individuals, there was no significant difference in thyroid hormone levels (FT3, FT4, and TSH). However, two patients developed thyroid toxicity, with increased levels of FT3, FT4 and a decreased level of TSH. Concomitantly, increased levels of thyroid autoantibodies were also observed in two other patients. Additionally, female participants showed significantly lower levels of FT3 and FT4 compared to the control group (p = 0.02 and p = 0.008, respectively). Among all of the COVID-19 survivors, a total of 10 patients (17%) showed deviations from the normal range of thyroid hormones and thyroid autoantibodies. The rest of the 48 patients (83%) revealed no abnormality. Conclusion: Female survivors experienced a significant change in thyroid functional status that lasted longer. However, further research needs to be performed on a larger scale before reaching a conclusion. Bangladesh J. Nuclear Med. 25(2): 93-97, 2022

Allium cameleon extract improved androgenesis and sexual behavior in male rat (Rattus norvegicus) via attenuation of oxidative damages increased sexual hormone and NO/cGMP cascade

Objective: Allium cameleon Cronquist (1981) (Liliaceae) is a bulb belonging to the garlic family, known as a medicinal plant used for its aphrodisiac activities in Cameroonian folk medicine. The present study was conducted to assess the aphrodisiac potentials of Allium cameleon aqueous extracts and investigate on the possible mechanisms by which this could be accomplished. Methods: The behavioral and biochemical tests were used to detect the aphrodisiac properties. The conceivable mechanisms of action of the aqueous extracts were investigated through determination of sexual hormones, nitric oxide, cGMP, and antioxidant activities in male rats treated with different plant extract concentrations. Results: Allium cameleon enhanced sexual excitement, improved semen quality, and the concentrations of testosterone, FSH and LH. Levels of endogenous antioxidant enzymes (CAT, SOD, and GSH) were improved. Penile nitric oxide and cGMP levels were boosted. Significant stimulation in sexual behavior, elevated spermatic production, raised of viability, morphology, count and sperm mobility were also observed. Treated animals exhibited elevated levels of endogenous antioxidant enzymes. Conclusion: These findings suggest that Allium cameleon possess aphrodisiac properties in rats that might be involve in the attenuation of oxidative damage, but also in the increase of sexual hormones production and NO/cGMP cascade.

PROPHYLAXIS COMPLEX OF PLACENTAL DYSFUNCTION AFTER THE INFLUENZA TRANSFERRED IN THE EARLY STAGES OF PREGNANCY

Aim of the research: to increase the effectiveness of tactics for the presence of placental dysfunction in the background of influenza by developing and implementing a pathogenetically sound set of treatment and prevention measures.&#x0D; Materials and methods. Test the effectiveness of the proposed set of treatment-and-prophylactic measures, 80 pregnant women were selected after influenza transferred to early pregnancy: 40 women whose pregnancies were optimized tactics (main group) and 40 women (comparison group), which were conducted according to clinical protocols of the Ministry of Health. The cytotoxicity of natural killers (NK) was assessed by flow cytometry. The level of cytokines in peripheral blood was studied by enzyme-linked immunosorbent assay: interleukin-1 (IL-1), interleukin-10 (IL-10). The study of 25-hydroxyvitamin D, 25- (OH) D was performed by enzyme-linked immunosorbent assay. Determination of serum hormones was performed using standard kits for immunoradiometric determination.&#x0D; Results. Homeostasis in the first trimester of pregnancy after influenza is characterized by disorders that reflect further plantar dysfunction: a list of NK cytotoxicity, proinflammatory changes in the cytokine profile, the procoagulant state of the hemostasis system. Optimization of conservation in such tactics is the use of pathogenetic and preventive measures, treatment and prevention measures to prevent placental insufficiency and reduce the negative effects of hyperimmune reactions (aspirin, dipyridamole, progestogens, intravenous immunoglobulin, vitamin D). The use of the recommended set of measures has a positive effect on hormonal functions of the placenta (normalization of progesterone and estradiol levels). Immunity also improved: one month after starting treatment with NK lysis 1:10 decreased from 24 [15; 30] to 17 [13; 21] (p &lt;0.05), the balance of the cytokine profile was manifested in a decrease in the ratio of IL-1 / IL-1 from 1.36 [1.12; 2.51] to 0.82 [0.42; 1.16] (p &lt;0.05). The proportion of women with suboptimal vitamin D levels decreased significantly from 87.5% to 37.5% (p &lt;0.05), and in cases of deficient conditions it was possible to bring the level of vitamin D in the blood to optimal or suboptimal. Reduction of Willebrand factor from 1.54 [1.38; 1.74] to 0.92 [0.68; 1.08] IU / ml (p &lt;0.05) indicates an improvement in endothelial function, which in turn creates favorable conditions for the development of the feto-placental complex. The use of the recommended management of pregnant women after influenza in early pregnancy has reduced the incidence of placental insufficiency by almost 2 times, which was diagnosed in 13 (35.0%) women in the main group against 25 (62.5%) patients in the comparison group p &lt;0.05).&#x0D; Conclusions. The use of the recommended set of measures has a positive effect on hormonal functions of the placenta (normalization of progesterone and estradiol levels), immune status (reduction of NK cytotoxicity, cytokine profile balance), elimination of vitamin D deficiency and reduction of procoagulation potential of hemostasis. Creating favourable conditions for the development of gestation can reduce the incidence of placental insufficiency by almost 2 times to 35.0% vs. 62.5%.

Development of a selective fluorescence derivatization strategy for thyroid hormones based on the Sonogashira coupling reaction

A new fluorescence derivatization technique for the determination of the thyroid hormones, 3,3′,5-triiodo-L-thyronine (T3, triiodothyronine) and 3,3′,5,5′-tetraiodo-L-thyronine (T4, L-thyroxine), in human serum was developed based on the Sonogashira coupling reaction. This derivatization reaction was recently utilized by our research group as a promising solution for the derivatization of ortho-substituted aryl halides that suffer from steric hindrance. T3 and T4 possess amino groups that could be derivatized by many reagents; however, these reagents are not useful in the case of biological analysis as they could non-selectively react with many biogenic amines and amino acids. Thus, herein we aimed at labeling the iodo-phenyl group of T3 and T4 as a selective fluorescence labeling approach suitable for biological analysis. The fluorescent alkyne, 2-(4-ethynylphenyl)-4,5-diphenyl-1H-imidazole (DIB-ET), can label the ortho-substituted aryl halides T3 and T4 in the presence of palladium and copper as catalysts, overcoming the steric hindrance of ortho-substitution. Furthermore, the application of the proposed method for the selective analysis of T3 and T4 in biological samples was successfully performed even in the presence of numerous biological components. The formed fluorescent derivatives produced from the reaction of DIB-ET and T3 and T4 could be determined by an HPLC system with fluorescence detection. The proposed method was successfully applied for the selective and sensitive determination of T3 and T4 in human serum with detection limits (S/N = 3) of 4.0 and 6.1 ng/mL and the recovery rate in the ranges of 84.3–92.1% and 81.3–84.9%, respectively. Therefore, the proposed method could be used as a new simple tool for the simultaneous determination of T3 and T4 in biological samples.

A One-Step Polyphenol Removal Approach for Detection of Multiple Phytohormones from Grape Berry

Phytohormones play an important role in regulating the maturation process and the quality-related metabolite accumulation of fruits, and their concentration variation has always been concerned during fruit development. However, berry fruits, such as grape berries, are rich in a large number of secondary metabolites, which brings great challenges to the isolation and determination of hormones. In this work, we used grapes as experimental materials and proposed a solid-phase extraction (SPE) protocol to efficiently isolate multiple hormones from phenol-rich matrix using a mixture of dichloromethane, methanol and formic acid as eluent. A highly sensitive method based on ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) was developed to quantify a total of 11 plant growth regulators, including the recognized phytohormones, in grape pericarp and seed. The established method showed satisfactory precision (RSD &lt; 11.3%) and linearity (R2 &gt; 0.9980). The limits of detection (LOD) and the limit of quantification (LOQ) were 0.001–0.75 ng/mL and 0.004–2.5 ng/mL, respectively. The recovery for the three levels of analytes spiked ranged from 63% to 118%, and the matrix effect was between 73% and 119%. Finally, the proposed method was applied to investigate the dynamic hormone concentration in Vitis vinifera L. cv. Cabernet Sauvignon berries from different vineyards, and assess the changes in endogenous hormones in grapes after treatment with exogenous growth regulators. We found that the contents of IP, ABA and IAA in pericarp and IP, IAA, IBA and SA in seed were significantly down-regulated after 10 days of treatment with NAA concentrations of 10 mg/L and 40 mg/L. In conclusion, this method helps to elucidate the role played by phytohormones in the maturation process and the accumulation of quality-related metabolites in phenol-rich fruits.

Parathormone concentration in the fluid of fine-needle parathyroid biopsy for assessing the effectiveness of surgical treatment of patients with primary hyperparathyroidism.

The aim of the work was to evaluate the usefulness of intraoperative determination of parathyroid hormone (PTH) concentration in the fluid of fine-needle biopsy lesions of pathologically parathyroid glands for their identification and evaluation of the effectiveness of surgical treatment of primary hyperparathyroidism (PHP). The study group comprised 75 patients: 65 women and 10 men, aged 33-78 years (mean 57.5), operated in the years 2019-2020 due to PHP. Seventy-nine biopsies of fine-needle aspiration lesions were assessed intraoperatively as pathological parathyroid glands were collected, and the concentration of PTH was determined in their lavage fluid. The results were compared with a control group of 20 patients operated at that time for various forms of goitre, without concomitant parathyroid disease. In 79 biopsies, the presence of the parathyroid gland was confirmed 72 times. The concentration of PTH in the biopsy fluid exceeded the value of 5000 pg/mL 70 times, 1 was 3733 pg/mL, and 1 was 1985 pg/mL. Seven times (8.9%), the biopsy was for a colloidal nodule instead of the parathyroid gland. The concentration of PTH in the fluids of these biopsies was low and ranged from 5.3 to 500 pg/mL. In the control group, the concentration of PTH in postoperative thyroid biopsies was low and did not exceed the reference value range of up to 64 pg/mL. Comparison of the results of the study group with the control group shows significantly higher PTH values in parathyroid biopsies compared to biopsies of thyroid specimens (p = 0.017), with a sensitivity of 100% and a specificity of nearly 93%. Intraoperative determination of PTH concentration in the fluid of fine-needle biopsy lesions with diseased parathyroid glands allows for their identification and evaluation of the effectiveness of surgical treatment of PHP. Testing the concentration of PTH in the biopsy fluid of dissected parathyroid glands is characterized by high sensitivity and specificity of the method. The value of PTH concentration in fluid from parathyroid biopsies does not depend on the cause of PHP.

P418 CARDIOMETABOLIC ALTERATIONS IN OBESE PATIENTS WITH OBSTRUCTIVE SLEEP APNEA SYNDROME

Abstract Background Obesity and Obstructive Sleep Apnea Syndrome (OSAS) has been recognized as two of the major cardiac risk factors and they often coexist in one patient. OSAS increase the risk of as myocardial infarction, heart failure and arrhythmias. On the other hand, obesity is and independent predictor of cardiovascular mortality since it causes insulin resistance, dyslipidemia, sympathetic hyperactivation, chronic inflammation and endothelial dysfunction. Aim To evaluate cardiometabolic profile, echocardiographic alterations and the presence of arrhythmias in obese patients with Obstructive Sleep Apnea Syndrome and non obese patients. Patients and Methods 67 obese patients (37 male/30 female) and 52 non obese patients (40 male/12 female) were enrolled. All subjects underwent anthropometrical evaluation and a venous blood sample for biochemical and hormonal determinations including fasting plasma glucose, fasting plasma insulin, Hba1c and OGTT to 5 hours. The insulin resistance was evaluated by HOMA–IR. All the patients underwent echocardiographic examination, polysomnographic evaluation, in order to diagnose the OSAS, and a simultaneous 12–lead Holter ECG to evaluate the presence of nocturnal arrhythmias. Results As expected, we found a better antropometrics profile in non obese patients. Obese patients display a severe OSAS compared to non–obese patient as shown by higher values of AHI (26.37 ±24.95 vs 16.58 ±14.07; P &amp;lt; 0.05), TC90 (19.61 ±24.45 vs 8.63 ±17.80; P &amp;lt; 0.05) and ODI (40.55 ±27.41 vs 21.06 ±22.78; P &amp;lt; 0,05). In obese patients we also found a significant reduction of both ejection fraction (55.54 ±7.97 vs 66.52 ±8.95; P &amp;lt; 0.05) and A–wave (0.75 ±0.17 vs 0.83 ±0.19; P &amp;lt; 0.05). In 20% of obese patients, 12–lead Holter ECG showed the presence of cardiac pause &amp;gt; 3 sec during apnea. In obese population alone we demonstrated a significant negative correlation between AHI and E–wave (r= –0.3; P = 0.04) and a positive correlation with interventricular septum (r = 0.38; P = 0.01) and left ventricular mass (r = 0.32; P = 0.02). Post–hoc analysis shown how these findings maintained statistical significance even when the obese patients were stratified in subgroups according to the severity of OSAS. Conclusions Our data, even if preliminary, seem to indicate that OSAS is not only linked to obesity but it also acts as a negative factor on the cardiometabolic risk.

MO033: Parathyroid hormone determination as an early predictor of progression in autosomal dominant polycystic kidney disease

Abstract BACKGROUND AND AIMS Total kidney volume (TKV) in patients with autosomal dominant polycystic kidney disease (ADPKD) is a known progression factor in these patients. Low levels of klotho and high levels of FGF23 in the blood of patients with ADPKD have been correlated with higher TKV even in early stages of the disease, even with glomerular filtration rate (eGFR) &amp;gt;90 mL/min/1.73 m2. Therefore, alterations in the levels of parathyroid hormone (PTH), vitamin D, tubular reabsorption of phosphates or urinary calcium could be correlated with the TKV and serve as an early serum biomarker of progression of kidney disease. METHOD Observational, cross-sectional, single-center study in a population of 87 patients under follow-up at our center during 2019–2020 for ADPKD. The TKV, obtained by the ellipsoid formula measured by magnetic resonance imaging (MRI) and the values of PTH, vitamin D, tubular phosphate reabsorption and urinary calcium were retrospectively collected. Subsequently, a statistical analysis of correlation between the different variables was performed. RESULTS Spearman's Rho correlation analysis obtained a positive correlation of moderate–strong intensity between the TKR of the sample and PTH levels (r = 0.638, P &amp;lt;0 0.0001) and a negative correlation of moderate intensity with tubular phosphate reabsorption (r = −0.479, P &amp;lt; 0.005). No statistically significant correlation was obtained between the TKV and the levels of urinary calcium or serum vitamin D. TKV was higher among patients with an eGFR &amp;gt;90 mL/min/1.73 m2 (CKD-EPI) with an elevated PTH (&amp;gt; 65 pg/mL), with a median of 530 mL (interquartile range 403–1184 mL), compared with the TKV of those patients with an eGFR &amp;gt;90 mL/min/1.73 m2 with normal PTH, whose median was 405 mL (interquartile range 258–576 mL). Despite this trend, the Mann–Whitney U test did not show a statistically significant difference between medians in the subgroup of patients with eGFR &amp;gt;90 mL/min/1.73 m2. However, the comparison of medians in the subgroup of patients with eGFR &amp;lt;90 mL/min/1.73 m2 was statistically significant with a P &amp;lt; 0.019, with higher TKR in those patients with high PTH. CONCLUSION The increase in PTH in patients with ADPKD has a strong positive correlation with TKV. Even with normal eGFR, elevated PTH is associated with a higher TKV, which could be an early serum biomarker of kidney disease progression in this group of patients.

Click Isotope Mass Probe for Highly Selective Determination of Trace Steroid Hormones in Food Samples.

Matrix effects are a great challenge for the quantitative analysis of complex food samples by liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS). Stable isotope labeling (SIL) has been widely used as an effective strategy to eliminate matrix effects. Herein, a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click-reaction-based SIL method was proposed for a highly sensitive and selective determination of six synthetic steroid hormones in three different food samples (milk, yogurt, and eggs) by high-performance liquid chromatography (HPLC)-MS/MS. A pair of novel SIL agents, N-(2-azidyl ethyl) aniline (d0-AEA) and d5-N-(2-azidyl ethyl) aniline (d5-AEA) were synthesized to label steroid hormones in the samples and standard solution, respectively. The reaction accomplishes in 30 min at 60 °C. The heavy labeled standards were used as internal standards (ISs), which experience the identical ionization processes with light labeled samples to minimize matrix effects. After derivatization, the ionization efficiencies of steroid hormones were greatly improved by 2-54-folds, and the matrix effects ranged from 88.6 to 99.8%. The established method achieved satisfactory detection limits (0.1-2.5 μg L-1) and high recoveries (85-102%). These results demonstrated that the proposed method holds unique advantages for trace steroid hormones analysis in foodstuffs.