Measurement of human urinary prolactin as a noninvasive study tool

Abstract We used a time-resolved solid-phase fluoroimmunoassay with a sensitivity of 25 ng/L on 40-fold-concentrated urines to measure urine prolactin (PRL) excretion. The nature of the immunoreactive material was verified to be PRL by: (a) column chromatography showing a monomeric 23-kDa peak; (b) similarity between fluoroimmunoassay and bioassay (Nb2 lymphoma cell) results; and (c) Western blot identification. In 20 normal subjects [serum PRL 6.8 (3.8-14.0) micrograms/L, median (and range)], urine PRL excretion was 0.15 (0.07-0.23) ng/h and 0.24 (0.15-0.54) micrograms/mol of creatinine. Urine values in seven hyperprolactinemic patients were all greater than the upper limit of normal. The correlation between urinary excretion rate and serum values was highly significant (r = 0.979; P < 0.001). These results indicate that a monomeric, immunologically reactive, biologically active form of PRL can be measured in urine at concentrations approximately 0.0005 that in serum. This urine PRL method may provide a practical tool for the repetitive, noninvasive study of PRL dynamics in field studies and in patients with reproductive disorders.

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Decreased circulating levels of active ghrelin are associated with increased oxidative stress in obese subjects

Acta Endocrinologica ◽

10.1530/eje.1.01977 ◽

2005 ◽

Vol 153 (3) ◽

pp. 403-407 ◽

Cited By ~ 34

Author(s):

Mina Suematsu ◽

Akira Katsuki ◽

Yasuhiro Sumida ◽

Esteban C Gabazza ◽

Shuichi Murashima ◽

...

Keyword(s):

Oxidative Stress ◽

Plasma Levels ◽

Active Form ◽

Inverse Correlation ◽

Normal Subjects ◽

Biologically Active ◽

Prostaglandin F ◽

Obese Subjects ◽

Circulating Levels ◽

And Oxidative Stress

Objective: To investigate the relationship between active ghrelin and oxidative stress in obese subjects. Design: We measured the plasma levels of free 8-epi-prostaglandin F2α (8-epi-PGF2α, a reliable and systemic marker of oxidative stress) and the active form of ghrelin in 17 obese and 17 normal subjects. The biologically active forms of ghrelin were measured using a commercially available radio-immunoassay kit and free 8-epi-PGF2α was measured using an enzyme immunoassay kit. Results: The circulating level of active ghrelin was significantly decreased (20.4 ± 2.6 vs 40.9 ± 3.9 fmol/ml, P < 0.01) while that of 8-epi-PGF2α was significantly increased (61.5 ± 9.6 vs 17.3 ± 3.4 pg/ml, P < 0.01) in obese subjects compared with normal subjects. The plasma levels of active ghrelin and 8-epi-PGF2α were significantly correlated in obese (r = −0.507, P < 0.05) and in all (r = −0.577, P < 0.01) subjects. Multivariate analysis showed that the plasma levels of active ghrelin and 8-epi-PGF2α were significantly and independently correlated in all subjects (F = 7.888, P < 0.01). Conclusions: There is an inverse correlation between circulating levels of active ghrelin and oxidative stress in obesity. Low circulating levels of active ghrelin may enhance oxidative stress and the process of atherosclerosis in obese subjects.

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Abnormal diurnal rhythm of plasma vasopressin and urinary output in patients with enuresis

AJP Renal Physiology ◽

10.1152/ajprenal.1989.256.4.f664 ◽

1989 ◽

Vol 256 (4) ◽

pp. F664-F671 ◽

Cited By ~ 13

Author(s):

S. Rittig ◽

U. B. Knudsen ◽

J. P. Norgaard ◽

E. B. Pedersen ◽

J. C. Djurhuus

Keyword(s):

Urinary Excretion ◽

Diurnal Rhythm ◽

Excretion Rate ◽

Normal Subjects ◽

Urinary Output ◽

Urinary Volume ◽

Urinary Excretion Rate ◽

Urinary Osmolality ◽

Plasma Vasopressin ◽

Nocturnal Increase

The diurnal variation of plasma vasopressin (AVP), urinary excretion rate, urinary osmolality, and serum osmolality was studied twice in 15 patients with enuresis and in 11 age, weight, and sex matched nonenuretic normal subjects. A diurnal rhythm of AVP with constant levels during the day (8 AM-10 PM) and a highly significant increase during the night (10 PM-8 AM) was found in normal subjects. In contrast, enuretics showed a significantly less pronounced nocturnal increase in AVP with significantly lower nocturnal levels than normal subjects. Normal subjects showed a diurnal rhythm in urinary excretion rate reciprocal to urinary osmolality with a low and highly concentrated nocturnal urinary output. In enuretics, however, this normal diurnal rhythm was absent. In conclusion, an abnormal diurnal rhythm of AVP seems to be an important pathophysiological factor in enuresis, explaining the abnormally high nocturnal urinary volume and the low nocturnal urinary osmolality found in these patients.

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Excretion of digoxin-like immunoreactivity in urine of normal subjects: correlations with excretion of creatinine and electrolytes.

Clinical Chemistry ◽

10.1093/clinchem/34.3.554 ◽

1988 ◽

Vol 34 (3) ◽

pp. 554-556 ◽

Cited By ~ 6

Author(s):

A Clerico ◽

O Giampietro ◽

G Gregori ◽

M G Del Chicca ◽

S Bertoli ◽

...

Keyword(s):

Urinary Excretion ◽

Excretion Rate ◽

Normal Subjects ◽

Urine Samples ◽

Urinary Excretion Rate ◽

The Mean ◽

Positive Correlations ◽

Excretion Rates ◽

Sodium And Potassium ◽

Normal Adults

Abstract To verify whether there is a variation in the 24-h urinary excretion of digoxin-like immunoreactivity (DLIS) in humans, we studied 18 normal adults, who collected their urines for 24-h in several portions. We then measured DLIS (by means of a sensitive RIA method), creatinine, sodium, and potassium concentrations in the urine samples. The mean urinary excretion rate for DLIS in the complete 24-h collection was 84.8 (SD 31.3) pg/min. The mean DLIS urinary excretion rate calculated for overnight collections was significantly lower than those of afternoon collections (P less than 0.01) and the 24-h collection (P less than 0.05). Significant positive correlations were found between urinary DLIS and excretion rates for creatinine (r = 0.347, P = 0.0016), Na+ (r = 0.232, P = 0.038), and K+ (r = 0.323, P = 0.003), respectively. Our data suggest that urinary excretion of DLIS is higher during "active" hours of the day, especially in the afternoon, than at rest, during the night.

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New immunoassay systems based on recombinant human lactoferrin

Doklady of the National Academy of Sciences of Belarus ◽

10.29235/1561-8323-2021-65-3-290-302 ◽

2021 ◽

Vol 65 (3) ◽

pp. 290-302

Author(s):

D. A. Semenov ◽

I. I. Vashkevich ◽

O. V. Sviridov

Keyword(s):

Solid Phase ◽

Food Additives ◽

Rabbit Antiserum ◽

Pharmaceutical Preparations ◽

Goat Milk ◽

Biologically Active ◽

Synthetic Methods ◽

Human Lactoferrin ◽

Time Resolved ◽

Recombinant Human Lactoferrin

In this work, soluble and solid phase immunoreagents, including recombinant human lactoferrin (rhLF), a complex of rhLF with europium ions, rabbit antiserum to rhLF, anti-rhLF immunoglobulin purified by antigen-affinity chromatography and the conjugates of this immunoglobulin with an Eu3+ chelate or horseradish peroxidase have been obtained by a combination of biochemical and synthetic methods using rhLF as an initial compound. Biospecific interactions of the reagents in four immunochemical systems were assessed by measuring the enzyme activity or time-resolved fluorescence. The study resulted in the development of fast and precise immunoassays for biologically active rhLF in transgenic goat milk and in protein fractions obtained in the course of pure rhLF manufacture, as well as in pharmaceutical preparations and food additives.

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Basic Studies of Various 99mTc-Labelled Renal Agents and Clinical Application of 99mTc-Malate

Nuklearmedizin ◽

10.1055/s-0037-1620604 ◽

1977 ◽

Vol 16 (01) ◽

pp. 36-41 ◽

Cited By ~ 1

Author(s):

T. Machida ◽

M. Miki ◽

M. Ueda ◽

A. Tanaka ◽

I. Ikeda

Keyword(s):

Excretion Rate ◽

Animal Experiments ◽

Imaging Agents ◽

Half Time ◽

Clinical Experiences ◽

Labelling Efficiency ◽

Urinary Excretion Rate ◽

Renal Imaging ◽

The Past ◽

Basic Studies

SummaryVarious renal imaging agents that were reported in the past and a new agent, 99mTc-malate as well as 99mTc-cystein acetazolamide complex were prepared using electrolysis and electrochemical methods. These were studied for their labelling efficiency. After animal experiments with selected 99mTc-com- pounds, 99mTc-rnalate proved to be sufficient for renal imaging with adequate concentration. 99mTcmalate differs from other renal imaging agents in the utilization of endogeneous metabolic product.The first half time of 99mTc-malate in humans is 17 minutes, on the average, and the urinary excretion rate of 99mTc-malate is 36±6.05% in 1 hour after intravenous administration, 44 ± 3.41% in 2 hours and 50 + 5.62% in 3 hours.In our 40 clinical experiences of 99m-Tc-rnalate, most cases demonstrated quite clear renal images in the serial scintiphotos except cases whose serum creatinines were over 4.5 mg/dl.

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Processing and Characterization of Recombinant von Willebrand Factor Expressed in Different Cell Types Using a Vaccinia Virus Vector

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648398 ◽

1992 ◽

Vol 67 (01) ◽

pp. 154-160 ◽

Cited By ~ 14

Author(s):

P Meulien ◽

M Nishino ◽

C Mazurier ◽

K Dott ◽

G Piétu ◽

...

Keyword(s):

Vaccinia Virus ◽

Von Willebrand Factor ◽

Human Fibroblasts ◽

Active Form ◽

Cell Types ◽

Biologically Active ◽

L Cells ◽

Von Willebrand ◽

Different Cell Types ◽

Willebrand Factor

SummaryThe cloning of the cDNA encoding von Willebrand factor (vWF) has revealed that it is synthesized as a large precursor (pre-pro-vWF) molecule and it is now clear that the prosequence or vWAgll is responsible for the intracellular multimerization of vWF. We have cloned the complete vWF cDNA and expressed it using a recombinant vaccinia virus as vector. We have characterized the structure and function of the recombinant vWF (rvWF) secreted from five different cell types: baby hamster kidney (BHK), Chinese hamster ovary (CHO), human fibroblasts (143B), mouse fibroblasts (L) and primary embryonic chicken cells. Forty-eight hours after infection, the quantity of vWF antigen found in the cell supernatant varied from 3 to 12 U/dl depending on the cell type. By SDS-agarose gel electrophoresis, the percentage of high molecular weight forms of vWF varied from 39 to 49% relative to normal plasma for BHK, CHO, 143B and chicken cells but was less than 10% for L cells. In all cell types, the two anodic subbands of each multimer were missing. The two cathodic subbands were easily detected only in BHK and L cells. By SDS-PAGE of reduced samples, pro-vWF was present in similar quantity to the fully processed vWF subunit in L cells, present in moderate amounts in BHK and CHO and in very low amounts in 143B and chicken cells. rvWF from all cells bound to collagen and to platelets in the presence of ristocetin, the latter showing a high correlation between binding efficiency and degree of multimerization. rvWF from all cells was also shown to bind to purified FVIII and in this case binding appeared to be independent of the degree of multimerization. We conclude that whereas vWF is naturally synthesized only by endothelial cells and megakaryocytes, it can be expressed in a biologically active form from various other cell types.

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Innovative approaches to the correction of chronotrope status of pregnant and lactating women

HEALTH OF WOMAN ◽

10.15574/hw.2016.114.37 ◽

2016 ◽

pp. 37-40

Author(s):

S.I. Zhuk ◽

◽

K.K. Bondarenko ◽

Keyword(s):

Folic Acid ◽

Neural Tube ◽

Active Form ◽

Biologically Active ◽

Lactating Women ◽

Pathological Conditions ◽

Prevention And Treatment ◽

The Impact ◽

High Bioavailability ◽

Key Words Mthfr

Most recent studies show the impact of violations in the metabolism of folate and metin period in the pathogenesis of neural tube defects (NTD) of the fetus. Metafolin has a number of advantages, which primarily includes direct intake of substances in biologically active form and the optimum effect, even in the case when the patient homozygote and/or heterozygote genotype 677С T polymorphism in MTHFR. With the aim of prevention and treatment of various pathological conditions related to folate deficiency during pregnancy, it is advisable to apply vitamin-mineral complexes, containing metafolin - active form of folate with high bioavailability. Key words: MTHFR, metafolin, folic acid, pregnancy.

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Synthetic Routes for 1,4-disubstituted 1,2,3-triazoles: A Review

Current Organic Chemistry ◽

10.2174/1385272823666190514074146 ◽

2019 ◽

Vol 23 (8) ◽

pp. 860-900 ◽

Cited By ~ 8

Author(s):

Chander P. Kaushik ◽

Jyoti Sangwan ◽

Raj Luxmi ◽

Krishan Kumar ◽

Ashima Pahwa

Keyword(s):

Solid Phase ◽

Click Reaction ◽

Biological Activities ◽

Triazole Ring ◽

Copper Catalysts ◽

Biologically Active ◽

Pharmaceutical Chemistry ◽

Biological Targets ◽

Synthetic Routes ◽

High Yields

N-Heterocyclic compounds like 1,2,3-triazoles serve as a key scaffolds among organic compounds having diverse applications in the field of drug discovery, bioconjugation, material science, liquid crystals, pharmaceutical chemistry and solid phase organic synthesis. Various drugs containing 1,2,3-triazole ring which are commonly available in market includes Rufinamide, Cefatrizine, Tazobactam etc., Stability to acidic/basic hydrolysis along with significant dipole moment support triazole moiety for appreciable participation in hydrogen bonding and dipole-dipole interactions with biological targets. Huisgen 1,3-dipolar azide-alkyne cycloaddition culminate into a mixture of 1,4 and 1,5- disubstituted 1,2,3-triazoles. In 2001, Sharpless and Meldal came across with a copper(I) catalyzed regioselective synthesis of 1,4-disubstituted 1,2,3-triazoles by cycloaddition between azides and terminal alkynes. This azide-alkyne cycloaddition has been labelled as a one of the important key click reaction. Click synthesis describes chemical reactions that are simple to perform, gives high selectivity, wide in scope, fast reaction rate and high yields. Click reactions are not single specific reaction, but serve as a pathway for construction of simple to complex molecules from a variety of starting materials. In the last few decades, 1,2,3-triazoles attracted attention of researchers all over the world because of their broad spectrum of biological activities. Keeping in view the biological importance of 1,2,3-triazole, in this review we focus on the various synthetic routes for the syntheisis of 1,4-disubstituted 1,2,3-triazoles. This review involves various synthetic protocols which involves copper and non-copper catalysts, different solvents as well as substrates. It will boost synthetic chemists to explore new pathway for the development of newer biologically active 1,2,3-triazoles.

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Folate in Red Rhapsody Strawberry—Content and Storage Stability.

Proceedings ◽

10.3390/foods_2020-07670 ◽

2020 ◽

Vol 70 (1) ◽

pp. 47

Author(s):

Julius Rami ◽

Caroline Dumler ◽

Nadine Weber ◽

Michael Rychlik ◽

Gabriele Netzel ◽

...

Keyword(s):

Storage Stability ◽

Active Form ◽

Food Composition ◽

Biologically Active ◽

Stable Isotope Dilution ◽

Isotope Dilution Assay ◽

C Storage ◽

Vitamin B9 ◽

Adults And Children ◽

And Storage

Folate (Vitamin B9) is critical for a range of biological functions in adults and children, including DNA, protein and neurotransmitter synthesis. It is also essential for the healthy development of the fetus in early pregnancy and for the prevention of neural tube defects, such as spina bifida. Strawberries are considered a tasty and healthy fruit consumed all over the world and may potentially be an important dietary source of natural folates. However, the relative importance of strawberry as a dietary source will depend on the total folate concentration, vitamer profile, storage stability and bioavailability to humans. Red Rhapsody, an important commercial strawberry cultivar in Australia, was screened for its folate content and storage stability by stable isotope dilution assay (SIDA). Total folate content ranged from 90–118 μg/100 g fresh weight (fw), which was well above the value in the Australian Food Composition Database (39 μg/100 g fw). 5-Methyltetrahydrofolate, the biologically active form in humans, was the principal vitamer present. Furthermore, folate remained relatively stable during refrigerated (4 °C) storage (loss of only 28% after 14 days of storage). This information is relevant for consumers since the inherent perishability of strawberry fruit makes refrigerated storage common practice in Australian households.

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