Relationship of advanced oxidative protein products in human saliva and plasma: age- and gender-related changes and stability during storage

Abstract The blood levels of advanced oxidative protein products (AOPP) elevate in aging and age-related diseases. However, saliva AOPP in healthy humans have been unexplored. Thus, we investigated 143 Chinese healthy adults to assay age- and gender-related changes in saliva and plasma AOPP levels and the stability of saliva AOPP stored both at -?20°C and -?80°C. We found the mean AOPP levels in saliva and plasma of 119 subjects were 7.51?±?3.20 and 28.31?±?5.53 μmol/L (μM). An age-dependent increase was observed in both saliva and plasma AOPP levels. This increase was particularly significant in the elderly subjects compared with that in the young and middle-aged ones. A significant positive correlation among age, saliva and plasma AOPP levels was observed. No gender-dependent difference was observed in either saliva or plasma AOPP levels during the aging process. Furthermore, AOPP levels in the 24 saliva samples showed no significant change at every successive determination during 4 weeks at -?80°C, whereas those levels significantly increased after 7 days of storage at -?20°C. These results indicate the feasibility to screen aging biochemical indicators using saliva AOPP as an alternative to blood AOPP. Saliva AOPP samples are suitable to be stored at -?80°C.     

Freeze-dried plasma proteins are stable at room temperature for at least 1 year

Thirty human EDTA plasma samples from male and female subjects ranging in age from 24 to 74 years were collected on ice, processed ice cold and stored frozen at ?80 °C, in liquid nitrogen (LN2), or freeze dried and stored at room temperature in a desiccator (FDRT) or freeze dried and stored at ?20 °C for 1 year (FD-20). In a separate experiment, EDTA plasma samples were collected onto ice, processed ice cold and maintained on ice ± protease inhibitors versus incubated at room temperature for up to 96 h. Random and independent sampling by liquid chromatography and tandem mass spectrometry (LC–ESI–MS/MS), as correlated by the MASCOT, OMSSA, X!TANDEM and SEQUEST algorithms, showed that tryptic peptides from complement component 4B (C4B) were rapidly released in plasma at room temperature. Random sampling by LC–ESI–MS/MS showed that peptides from C4B were undetectable on ice, but peptides were cleaved from the mature C4B protein including NGFKSHALQLNNR within as little as 1 h at room temperature. The frequency and intensity of precursors within ± 3 m/z of the C4B peptide NGFKSHALQLNNR was confirmed by automated targeted analysis where the precursors from MS/MS spectra that correlated to the target sequence were analyzed in SQL/R. The C4B preproprotein was processed at the N terminus to release the mature chain that was cleaved on the carboxyl side of the isoprene C2 domain within a polar C terminal sequence of the mature C4B protein, to reveal the thioester reaction site, consistent with LC–ESI–MS/MS and Western blot. Random sampling showed that proteolytic peptides from complement component C4B were rarely observed with long term storage at ? 80 °C in a freezer or in liquid nitrogen (LN2), freeze drying with storage at ? 20 °C (FD-20 °C) or freeze drying and storage at room temperature (FDRT). Plasma samples maintained at room temperature (RT) showed at least 10-fold to 100-fold greater frequency of peptide correlation to C4B and measured peptide intensity compared to samples on ice for up to 72 h or stored at ? 80 °C, LN2, FDRT or FD-20 °C for up to a year.     

Proteomic Studies of Saliva: A Proposal for a Standardized Handling of Clinical Samples

Background

In recent years, differential analysis of proteins from human saliva, i.e., proteomic analysis, has received much attention mainly due to its unstressful sampling and its great potential for biomarker research. It is widely considered that saliva is a highly stable medium for proteins thanks to a large amount of antiprotease agents, even at ambient and physiological temperatures.

Objective

To find the best protocol for the handling of samples, we have investigated the stability of saliva proteins stored at different temperatures (from ?80 to 20°C) by one- and two-dimensional electrophoresis.

Results

At 20°C, no major changes were observed on protein one-dimensional profiles following 1 day of storage; however, between 7 days and 30 days, the native alpha-amylase band decreased slightly to give several bands with molecular weight between 35 and 25 kDa. The same phenomenon appeared after 30 days of storage at 4°C. Two-dimensional analysis of salivary maps revealed degradation from day 7 of several protein groups for samples stored at 20°C.

Conclusion

All these findings have to be carefully considered when saliva is collected for clinical proteomic analysis. We can conclude that, to maintain the optimum stability of saliva proteins, saliva samples should be collected on ice followed by the addition of protease inhibitor cocktail, centrifuged to remove insoluble material, and stored at ?20 or ?80°C.     

Influence of Trace Elements on Stabilization of Aqueous Solutions of Ascorbic Acid

Together with vitamin C, zinc, selenium, manganese, and magnesium play a vital role in the preservation of organs scheduled for transplantation. In the present study, it is shown that addition of 1?mg/l of these elements influences the stability of 0.3?mM ascorbic acid solutions. The solution??s stability was estimated using an accelerated stability test. The concentration of vitamin C was measured using a validated spectrophotometric method, which uses the reduction of 2,6-dichlorophenoloindophenol by ascorbic acid. Elevated temperatures, the factor accelerating substances?? decomposition reaction rate, were used in the tests. The research was conducted at two temperatures at intervals of 10?°C: 80?±?0.1 and 90?±?0.1?°C. It was stated that the studied substances?? decomposition occurred in accordance with the equation for first-order reactions. The function of the logarithmic concentration (log%C) over time was revealed to be rectilinear. This dependence was used to determine the kinetics of decomposition reaction rate parameters. The stabilization of vitamin C solutions was measured as the time in which 10?% of the substance decomposed at 20 and 0?°C. Addition of Se(IV) or Mg(II) ions significantly increase the stability of ascorbic acid solution (??34 and ??16?%, respectively), but Zn(II) causes a significant decrease in stability by ??23?%. Addition of Mn(II) has no significant influence on vitamin C stability.     

Long-term stability of parameters of antioxidant status in human serum

Abstract

The antioxidant status of serum or plasma can be determined using several commercially available assays. Here, four different assays, total antioxidant status (TAS), its second-generation assay (TAS2), biological antioxidant potential (BAP), and enzymatic assay using horseradish peroxidase (EAOC), were applied on human serum samples to test the temperature stability of antioxidants, upon storage of serum for 12 months. The two or three most commonly used temperatures for storage, that is, ? 20, ? 70 (or ? 80), and ? 196°C, were selected. The general conclusion is that all assays were stable at the temperatures tested. In addition, there were almost no statistically significant differences between the samples stored at different temperatures. Only the rank order of the EAOC assay was not very good in samples stored at ? 20°C. Also three components contributing to the total antioxidant capacity, uric acid, creatinine and bilirubin, showed no statistically significant differences between the temperatures. Therefore, storage at ? 20°C is sufficient to maintain a proper assay outcome of most of the total antioxidant assays, although storage at ? 70/80°C is to be preferred for longer storage times.     

A comparison of native and non-urate Total Antioxidant Capacity of fasting plasma and saliva among middle-aged and older subjects

Objectives: As plasma and salivary total antioxidant capacity (TAC) is mainly contributed by uric acid (UA), the present study measures non-urate TAC (Nu-TAC). The aim of the study was to correlate plasma native TAC, Nu-TAC and UA with their salivary analogues, and compare the UA contribution in both body fluids using two different methods.

Methods: The study involved 55 middle-aged and older subjects (66.7?±?4.5 years). TAC was determined simultaneously with two methods (ferric reducing ability of plasma – FRAP, 2.2-diphenyl-1-picryl-hydrazyl – DPPH and countertypes for saliva – FRAS and DPPHS test), with and without UA (native TAC and Nu-TAC, respectively). Plasma UA and salivary UA (SUA) were assessed.

Results: Subjects with increased FRAP, DPPH and UA had higher FRAS, DPPHS and SUA, respectively (P?P?Discussion: Our findings suggest that saliva is a good predictor for native plasma TAC but not for Nu-TAC. UA level is comparably dominant in saliva and in plasma according to DPPH, but lower in plasma according to FRAP.     

Effects of storage conditions on the stability and distribution of clinical trace elements in whole blood and plasma: Application of ICP-MS

BackgroundKnowledge of trace element stability during sample handling and preservation is a prerequisite to produce reliable test results in clinical trace element analysis.MethodAn alkaline dissolution method has been developed using inductively coupled plasma mass spectrometry to quantify eighteen trace element concentrations: vanadium, chromium, manganese, cobalt, nickel, copper, zinc, arsenic, selenium, bromine, molybdenum, cadmium, antimony, iodine, mercury, thallium, lead, and bismuth in human blood, using a small sample volume of 0.1 mL. The study evaluated the comparative effects of storage conditions on the stability of nutritionally essential and non-essential elements in human blood and plasma samples stored at three different temperatures (4 °C, −20 °C and −80 °C) over a one-year period, and analysed at multiple time points. The distribution of these elements between whole blood and plasma and their distribution relationships are illustrated using blood samples from 66 adult donors in Queensland.ResultsThe refrigeration and freezing of blood and plasma specimens proved to be suitable storage conditions for many of the trace elements for periods up to six months, with essentially unchanged concentrations. Substantially consistent recoveries were obtained by preserving specimens at −20 °C for up to one year. Ultra-freezing of the specimens at −80 °C did not improve stability; but appeared to result in adsorption and/or precipitation of some elements, accompanied by a longer sample thawing time. A population sample study revealed significant differences between the blood and plasma concentrations of six essential elements and their relationships also varied significantly for different elements.ConclusionBlood and plasma specimens can be reliably stored at 4 °C for six months or kept frozen at −20 °C up to one year to obtain high quality test results of trace elements.     

Effect of storage time and temperature on steroid and protein hormone concentrations in porcine plasma and serum

The effect of storage time and temperature of porcine blood prior to quantitation of hormone concentrations by radioimmunoassay (RIA) was evaluated. Blood from each of four luteal phase gilts was used to determine cortisol (CS) and progesterone (P) concentrations, while blood from each of four ovariectomized gilts and each of four lactating sows was used to determine luteinizing hormore (LH) and prolactin (PRL) concentrations, respectively. Blood was collected via jugular puncture from each animal within a 30-sec time period and placed into 18 heparinized and 18 nonheparinized tubes. One sample with and without heparin was stored in ice water (4°C) or at 28°C for 0.25, 2, 4, 6, 8, 12, 24, 36 or 48 hours. After storage, blood was centrifuged at 4°C and plasma or serum was collected and stored at ?20°C until quantitated by RIA. There were no differences (P>0.05) between plasma and serum concentrations (X ± SE, ng/ml) of CS (26.9 ± 0.8 vs 28.5 ± 0.8), P (24.7 ± 0.7 vs 24.8 ± 0.8), LH (2.1 ± 0.1 vs 2.2 ± 0.1) or PRL (53.2 ± 2.3 vs 52.6 ± 2.1). Similarly, storage temperature (4 vs 28°C) did not affect the concentrations of P (25.7 ± 0.8 vs 23.9 ± 0.7), LH (2.2 ± 0.1 vs 2.2 ± 0.1) or PRL (53.7 ± 2.1 vs 53.2 ± 2.3). Howver, CS concentrations decreased (P<0.05) from 28.5 ± 0.5 (4°C) to 26.9 ± 0.8 ng/ml (28°C). There was an animla x time interaction for CS concentration when plasma and serum were stored at both 4°C (P<0.001) and 28°C (P<0.003). There was also and animal x time interaction (P<0.03) for LH concentrations. The P and PRL concentrations decreased linearly by 0.0615 ng/hr (P<0.001) and 0.0625 ng/hr (P<0.004), respectively, with increased storage time.     

Preservation of Phakopsora pachyrhizi Uredospores

This study compared different temperatures and dormancy‐reversion procedures for preservation of Phakopsora pachyrhizi uredospores. The storage temperatures tested were room temperature, 5°C, ?20°C and ?80°C. Dehydrated and non‐dehydrated uredospores were used, and evaluations for germination (%) and infectivity (no. of lesions/cm²) were made with fresh harvested spores and after 15, 29, 76, 154 and 231 days of storage. The dormancy‐reversion procedures evaluated were thermal shock (40°C/5 min) followed or not by hydration (moist chamber/24 h). Uredospores stored at room temperature were viable only up to a month of storage, regardless of their hydration condition. Survival of uredospores increased with storage at lower temperatures. Dehydration of uredospores prior to storage increased their viability, mainly for uredospores stored at 5°C, ?20°C and ?80°C. At 5°C and ?20°C, dehydrated uredospores showed increases in viability of at least 47 and 127 days, respectively, compared to non‐dehydrated spores. Uredospore germination and infectivity after storage for 231 days (7.7 months), could only be observed at ?80°C, for both hydration conditions. At this storage temperature, dehydrated and non‐dehydrated uredospores exhibited 56 and 28% of germination at the end of the experiment, respectively. Storage at ?80°C also maintained uredospore infectivity, based upon levels of infection frequency, for both hydration conditions. Among the dormancy‐reversion treatments applied to spores stored at ?80°C, those involving hydration allowed recoveries of 85 to 92% of the initial germination.     

Stability of Solubilized Benzodiazepine Receptors

ABSTRACT

According to the observations of other researchers, benzodiazepine receptors solubilized with sodium deoxycholate are unstable, but stability can be improved by exchanging deoxycholate for Triton X-100. In our experiments we conclude that the choice of detergent is not the restrictive factor for the stability of the solubilized receptors, but the storage conditions are. Solubilized receptors, either stored in sodium deoxycholate or in Triton X-100, were stable for at least 2 months when stored at ?80°C, but both preparations were strongly unstable when stored at ?20°C. Alternatively, sodium deoxycholate-solubilized receptors may be lyophilized and then stored at ?20°C.     

Long-term stability of oxidative stress biomarkers in human serum

The storage time and storage temperature might affect stability of oxidative stress biomarkers, therefore, they have to be analyzed after long-term storage of serum samples. The stability of three biomarkers reflecting oxidative stress: reactive oxygen metabolites (ROM) for hydroperoxides, total thiol levels (TTL) for the redox status and biological antioxidant potency (BAP) for the antioxidant status, was investigated at several time points during 60 months of storage at ?20 and ?80?°C. Biomarkers ROM and BAP showed a very good stability during storage for 60 months at both temperatures. In addition, the correlation of the data after 60 months of storage compared with the starting data was very good with correlation coefficients >0.9. The TTL assay showed good results in serum samples stored at ?80?°C, but not in samples stored at ?20?°C. Serum samples for analysis of the set of oxidative stress biomarkers ROM, BAP and TTL can be stored up to 60 months at ?80?°C. ROM and BAP can also be stored at ?20?°C during this period. The present results are very important for the biomarker-related epidemiological studies that make use of biobanks with samples stored for many years and for new project planning, including sample storage conditions.     

The stability of almond β-glucosidase during combined high pressure–thermal processing: a kinetic study

The thermal and the combined high pressure–thermal inactivation kinetics of almond β-glucosidase (β-d-glucoside glucohydrolase, EC 3.2.1.21) were investigated at pressures from 0.1 to 600 MPa and temperatures ranging from 30 to 80 °C. Thermal treatments at temperatures higher than 50 °C resulted in significant inactivation with complete inactivation after 2 min of treatment at 80 °C. Both the thermal and high pressure inactivation kinetics were described well by first-order model. Application of pressure increased the inactivation kinetics of the enzyme except at moderate temperatures (50 to 70 °C) and pressures between 0.1 and 100 MPa where slight pressure stabilisation of the enzyme against thermal denaturation was observed. The activation energy for the inactivation of the enzyme at atmospheric pressure was estimated to be 216.2?±?8.6 kJ/mol decreasing to 55.2?±?3.9 kJ/mol at 600 MPa. The activation volumes were negative at all temperature conditions excluding the temperature–pressure range where slight pressure stabilisation was observed. The values of the activation volumes were estimated to be ?29.6?±?0.6, ?29.8?±?1.7, ?20.6?±?3.2, ?41.2?±?4.8, ?36.5?±?1.8, ?39.6?±?4.3, ?31.0?±?4.5 and ?33.8?±?3.9 cm³/mol at 30, 35, 40, 45, 50, 60, 65 and 70 °C, respectively, with no clear trend with temperature. The pressure–temperature dependence of the inactivation rate constants was well described by an empirical third-order polynomial model.     

Variability of salivary markers of oxidative stress and antioxidant status in young healthy individuals

Objectives: Salivary advanced glycation end-products (AGEs), advanced oxidation protein products (AOPP), total antioxidant capacity (TAC), and ferric reducing ability of saliva (FRAS) are increased in various diseases. Little data exist for these markers in the healthy population. The aim of this study was to assess the inter-individual and intra-individual variability of AGEs, AOPP, TAC, and FRAS in the saliva of young healthy individuals.

Methods: Unstimulated saliva samples were collected from 16 females and 18 males daily over a period of 30 days. Markers were measured using spectrophotometric and spectrofluorometric microplate-based methods.

Results: All salivary markers measured were significantly higher in men than in women (P?<?0.05 for AGEs; P?<?0.001 for AOPP, TAC, and FRAS). The inter-individual variability was approximately 60% for AGEs and AOPP and 30–40% for TAC and FRAS in both genders. The inter-individual variability of FRAS was higher in men vs. women (P?<?0.01). Intra-individual variability ranged from 20% for TAC, to 30% for AGES and FRAS and 45% for AOPP.

Discussion: Intra-individual variability of salivary AGEs, AOPP, TAC, and FRAS indicates that their use is currently limited to large cohort studies. Identifying the underlying factors related to the high inter-individual and intra-individual variability is needed. Sex differences should be considered in future studies.     

Effect of freezing and thawing on the biomechanical characteristics of porcine ocular tissues

PurposeTo evaluate the effect of freezing and thawing on the biomechanical properties of ex-vivo porcine ocular tissue.MethodsThirty-six porcine eyes (18 pairs) were obtained fresh from a local abattoir and split into two groups of nine pairs to study the effect of storage at −20 °C and −80 °C. A randomly-selected eye from each pair (Control Group, CG) was tested fresh while the fellow eyes were frozen for 14 days, either at −20 °C and −80 °C (Frozen Group, FG) before thawing and testing. Seventy-two strips were extracted from the corneas and scleras of eye globes and subjected to uniaxial tension tests under loads up to 1.0 N. Following five preconditioning cycles, the load and elongation data obtained experimentally were analysed to derive the tissue’s stress-strain and tangent modulus-strain behaviour.ResultsCorneal tissue subjected to freezing at −20 °C exhibited significant increases in tangent modulus (mechanical stiffness) by 13 ± 17% (p = 0.003) at 1% strain and 14 ± 12% (p < 0.001) at 2% strain. In contrast, the increases in corneal stiffness at −80 °C were insignificant (6 ± 14%, p = 0.099 at 1% strain, 6 ± 15%, p = 0.091 at 2% strain). The corresponding increases in tangent modulus in the sclera were all insignificant (for −20 °C: 4 ± 14%, p = 0.265 at 1% strain, 3 ± 9%, p = 0.186 at 2% strain; for −80 °C: 3 ± 18%, p = 0.537 at 1% strain and 3 ± 18%, p = 0.491 at 2% strain).ConclusionsThe study provided evidence that freezing and thawing led to insignificant changes in ocular tissue stiffness except in corneal tissue that was frozen at −20 °C.     

Determination of the antihypertensive agent 1-(2-aminoethyl)-3-(2,6-Dichlorophenyl)thiourea in plasma and urine by high-performance liquid chromatography

A rapid, sensitive and specific normal-phase (adsorption) high-performance liquid chromatographic (HPLC) assay was developed for the determination of 1-(2-aminoethyl)-3-(2,6-dichlorophenyl)thiourea [I] in plasma and urine. The assay involves the extraction of the compound into methylene chloride from plasma or urine buffered to pH 10, and the HPLC analysis of the residue dissolved in methylene chloride—methanol—heptane (85:10:5). A 10-μm silica gel column was used with methylene chloride—methanol—heptane—ammonium hydroxide (85:10:5:0.1) as the eluting solvent. The effluent was monitored at 254 nm and quantitation was based on the peak height vs. concentration technique. The assay has a recovery of 64.5 ± 4.5% (S.D.) from plasma and 96.0 ± 6.3% (S.D.) from urine in the concentration range of 0.1–2 μg per ml and 2–40 μg per 0.1 ml of plasma and urine, respectively, with a limit of detection of 0.05–0.1 μg [I] per ml of plasma using a 1-ml specimen and 0.1 μg per ml urine using a 0.1-ml specimen, respectively. The assay was applied to the determination of plasma levels and urinary excretion of the compound [I] in dog following the oral administration of 28.8 mg of [I] · maleate per kg body weight.The HPLC assay was also used to determine the stability of [I] and for the measurement of a potential degradation product, clonidine [II] [2-(2,6-dichlorophenylamino)-2-imidazoline] in pooled human plasma stored at ?17°C, and pooled human urine stored at ?17°C and ?90°C, respectively.     

Attenuated thermoregulatory responses with increased plasma osmolality in obese subjects during two seasons

Obese subjects may be more vulnerable to injury from heat stress, and appear to be less efficient at thermoregulation. Sweat rate, tympanic temperature and osmolality in obese subjects were investigated in Japan during two seasons. The purpose of this study was to examine the relationship between obesity, thermoregulatory response and season. Five obese (BMI, 32.0?±?4.9 kg/m²) and five non-obese (BMI, 23.2?±?2.9 kg/m²) men participated in this experiment at latitude 35°10′ N and longitude 136°57.9′E. The average atmospheric temperature was 29.1?±?1.0 °C in summer and 3.3?±?1.4 °C in winter. Tympanic temperature and sweat rate were measured during leg water immersion at 42 °C for 30 min. Blood samples were analyzed for plasma osmolality. The relationship between tympanic temperature and sweat rate decreased significantly in obese compared to in non-obese subjects in both seasons, there being a lowered sweat rate for any core temperature in obese subjects. Plasma osmolality was significantly higher in obese than in non-obese subjects in both seasons. Thermal sensation increased significantly in non-obese than in obese in winter but not in summer. Our data show that thermoregulatory responses are attenuated in obese subjects compared with controls, suggesting that obese people are at increased risk of heat-related illnesses.     

Immobilization and characterization of lipase from an indigenous Bacillus aryabhattai SE3-PB isolated from lipid-rich wastewater

Abstract

Extracellular lipase from an indigenous Bacillus aryabhattai SE3-PB was immobilized in alginate beads by entrapment method. After optimization of immobilization conditions, maximum immobilization efficiencies of 77%?±?1.53% and 75.99%?±?3.49% were recorded at optimum concentrations of 2% (w/v) sodium alginate and 0.2?M calcium chloride, respectively, for the entrapped enzyme. Biochemical properties of both free and immobilized lipase revealed no change in the optimum temperature and pH of both enzyme preparations, with maximum activity attained at 60?°C and 9.5, respectively. In comparison to free lipase, the immobilized enzyme exhibited improved stability over the studied pH range (8.5–9.5) and temperature (55–65?°C) when incubated for 3?h. Furthermore, the immobilized lipase showed enhanced enzyme-substrate affinity and higher catalytic efficiency when compared to soluble enzyme. The entrapped enzyme was also found to be more stable, retaining 61.51% and 49.44% of its original activity after being stored for 30 days at 4?°C and 25?°C, respectively. In addition, the insolubilized enzyme exhibited good reusability with 18.46% relative activity after being repeatedly used for six times. These findings suggest the efficient and sustainable use of the developed immobilized lipase for various biotechnological applications.     

Human blood plasma advanced oxidation protein products (AOPP) correlates with fibrinogen levels

In 1996 a novel oxidative stress biomarker, referred to as advanced oxidation protein products (AOPP) was detected in the plasma of chronic uremic patients. The aim of the present studies was to find out that which plasma fraction(s) is responsible for AOPP reactivity. Thermal treatment of pooled samples of human citrate-plasma or EDTA-plasma at 50°C resulted in a rapid and parallel loss of fibrinogen concentration and AOPP reactivity. On the basis of time course and t_1/2 values following thermal treatment, AOPP was indistinguishable from fibrinogen. There was a statistically significant (p < 0.0001) correlation between levels of blood plasma fibrinogen and AOPP in patients (n = 61) with various peripheral vascular or cardiovascular diseases. There was also a significant (p < 0.0001) relationship between plasma levels of fibrinogen and molar AOPP/fibrinogen ratio indicating that higher fibrinogen concentrations were associated with more oxidatively transformed groups on the molecule. Results of the present studies suggest that post-translationally modified fibrinogen is a key molecule responsible for human plasma AOPP reactivity. It remains to be elucidated what is the pathophysiological significance of the post-translationally modified fibrinogen in the inflammation-associated events of atherosclerosis, in platelet aggregation, and as a cardiovascular risk biomarker.     

The Influences of Chromium Supplementation on Glycemic Control,Markers of Cardio-Metabolic Risk,and Oxidative Stress in Infertile Polycystic ovary Syndrome Women Candidate for In vitro Fertilization: a Randomized,Double-Blind,Placebo-Controlled Trial

This study was carried out to investigate the effects of chromium intake on glycemic control, markers of cardio-metabolic risk, and oxidative stress in infertile polycystic ovary syndrome (PCOS) women candidate for in vitro fertilization (IVF). This randomized double-blind, placebo-controlled trial was done among 40 subjects with infertile PCOS candidate for IVF, aged 18–40 years old. Individuals were randomly allocated into two groups to take either 200 μg/day of chromium (n?=?20) or placebo (n?=?20) for 8 weeks. Biochemical parameters were assessed at baseline and at end-of-trial. Compared with the placebo, taking chromium supplements led to significant reductions in fasting plasma glucose (??2.3?±?5.7 vs. +?0.9?±?3.1 mg/dL, P?=?0.03), insulin levels (??1.4?±?2.1 vs. +?0.4?±?1.7 μIU/mL, P?=?0.004), homeostatic model of assessment for insulin resistance (??0.3?±?0.5 vs. +?0.1?±?0.4, P?=?0.005), and a significant increase in quantitative insulin sensitivity check index (+?0.004?±?0.008 vs. ??0.001?±?0.008, P?=?0.03). In addition, chromium supplementation significantly decreased serum triglycerides (??19.2?±?33.8 vs. +?8.3?±?21.7 mg/dL, P?=?0.004), VLDL- (??3.8?±?6.8 vs. +?1.7?±?4.3 mg/dL, P?=?0.004) and total cholesterol concentrations (??15.3?±?26.2 vs. ??0.6?±?15.9 mg/dL, P?=?0.03) compared with the placebo. Additionally, taking chromium supplements was associated with a significant increase in plasma total antioxidant capacity (+?153.9?±?46.1 vs. ??7.8?±?43.9 mmol/L, P?<?0.001) and a significant reduction in malondialdehyde values (?0.3?±?0.3 vs. +?0.1?±?0.2 μmol/L, P?=?0.001) compared with the placebo. Overall, our study supported that chromium administration for 8 weeks to infertile PCOS women candidate for IVF had beneficial impacts on glycemic control, few variables of cardio-metabolic risk, and oxidative stress.