Peruvian Maca (Lepidium peruvianum) - III: The Effects of Cultivation Altitude on Phytochemical and Genetic Differences in the Four Prime Maca Phenotypes.

In two trials, dietary and Glucosinolates' characteristics in four Maca phenotypes have been examined with an extension into the determination of DNA sequences. Hypocotyls of the four prime phenotypes of Peruvian Maca - Lepidium peruvianum Chacon, labelled as "Yellow", "Black", "Red" and "Purple" were separated from mixed Maca crops cultivated in four geographically-distant locations in the Peruvian Andes at altitudes between 2,800m and 4,300 m a.s.l. It was found that at higher altitudes where Red and Purple Maca phenotypes were grown, the significantly higher (P<0.05) Glucosinolates' concentrations, adopted as the marker of Maca physiological activity, were observed with the Purple phenotype showing the highest Glucosinolates' content at 4,300m a.s.l., followed by the Red-coloured hypocotyls. Black Maca showed a reversal, but also a significant (P<0.05) trend, while the Yellow phenotype showed no visible altitude-inflicted response (P>0.05) and has consistently the lowest Glucosinolates content. Thus, it is reasonable to assume that the altitude at which Red, Purple and Black phenotypes of L. peruvianum are grown, may be responsible for the variation in physiologic functionalities, leading to different than expected specific therapeutic and health benefits induced by Maca phenotypes grown at diverse altitudes. Although promising, insufficiently precise differences in DNA sequences failed to distinguish, without any reasonable doubt, four Maca phenotypes cultivated either in the same or geographically-distant locations, and harvested at different altitudes a.s.l. Further research on DNA sequences is needed, with more primers and larger number of Maca phenotypes, considering biosynthesis of secondary metabolites and adaptation pathways induced by harsh environment at altitudes where Maca is cultivated.

Oleanolic Acid A-lactams Inhibit the Growth of HeLa, KB, MCF-7 and Hep-G2 Cancer Cell Lines at Micromolar Concentrations.

Oleanolic acid ketones, oximes, lactams and nitriles were obtained. Complete spectral characterizations (IR, (1)H NMR, (13)C NMR, DEPT and MS) of the synthesized compounds are presented. The derivatives had oxo, hydroxyimino, lactam or nitrile functions at the C-3 position, an esterified or unmodified carboxyl group at the C- 17 location and, in some cases, an additional oxo function at the C-11 position. The new compounds were tested for cytotoxic activity on the HeLa, KB, MCF-7 and Hep-G2 cancer cell lines with the application of MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test. Among the tested compounds, some oximes and all lactams proved to be the most active cytotoxic agents. These triterpenes significantly inhibited the growth of the HeLa, KB, MCF-7 and Hep-G2 cancer cell lines at micromolar concentrations.

Improvement in Long-Term Memory following Chronic Administration of Eryngium planum Root Extract in Scopolamine Model: Behavioral and Molecular Study.

Eryngium planum L. (EP) is as a rare medicinal plant with a lot of potentials as pharmaceutical crops. The aim of our study was to assess the effect of subchronic (28-fold) administration of a 70% ethanol extract of EP roots (200 mg/kg, p.o.) on behavioral and cognitive responses in Wistar rats linked with acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and beta-secretase (BACE-1) mRNA levels and AChE and BuChE activities in the hippocampus and frontal cortex. On the last day of experiment, 30 min after the last dose of EP or Huperzine A (HU), scopolamine (SC) was given at a dose of 0.5 mg/kg b.w. intraperitoneally. The results of a passive avoidance test showed an improvement in long-term memory produced by the EP extract in both scopolamine-induced rats and control group. EP caused an insignificant inhibition of AChE and BuChE activities in the frontal cortex and the hippocampus. EP decreased mRNA AChE, BuChE, and BACE-1 levels, especially in the cortex. Our results suggest that the EP extract led to the improvement of the long-term memory in rats coupled with total saponin content. The mechanism of EP action is probably complicated, since HPLC-MS analysis showed 64 chemical compounds (phenolics, saponins) in the extract of EP roots.

Advancing the delivery of anticancer drugs: Conjugated polymer/triterpenoid composite.

Exemplifying the synergy of anticancer properties of triterpenoids and ion retention qualities of conjugated polymers, we propose a conducting matrix to be a reservoir of anticancer compounds. In this study, poly(3,4-ethylenedioxythiophene), PEDOT, based matrix for electrically triggered and local delivery of the ionic form of anticancer drug, oleanolic acid (HOL), has been investigated. An initial, one-step fabrication procedure has been proposed, providing layers exhibiting good drug release properties and biological activity. Investigation of obtained systems and implementation of modifications revealed another route of fabrication. This procedure was found to yield layers possessing a significantly greater storage capacity of OL(-), as evidenced by the 52% increase in the drug concentrations attainable through electro-assisted release. Examination of the biological activity of immobilised and released OL(-) molecules proved that electrochemical treatment has negligible impact on the anticancer properties of OL(-), particularly when employing the three-step procedure, in which the range of applied potentials is limited. PEDOT/OL(-) composite has been demonstrated to be a robust and cost-effective material for controlled drug delivery.

The influence of mianserin on TNF-α, IL-6 and IL-10 serum levels in rats under chronic mild stress.

BACKGROUND: Antidepressants are known to affect the immunological system through mechanisms which are not completely understood. The aim of the present study was to evaluate the effect of the atypical antidepressant mianserin on the levels of tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6) and interleukin-10 (IL-10) in the blood of rats in an experimental model of depression. METHODS: Male Wistar rats were subjected to chronic mild stress (CMS) according to Willner's method for 6 weeks. Following the development of anhedonia, the stressed and control rats (non-stressed animals) were treated with mianserin (10 mg/kg ip, twice daily) for three weeks. On the last day of the experiment, a lipopolysaccharide (LPS, 100 µg/kg ip) was injected to mianserin- or vehicle-treated rats. TNFα, IL-6 and IL-10 levels in the blood of the rats were assayed using ELISA methods. RESULTS: The results indicated a significantly increased TNFα level in stressed animals when compared with the non-stressed (control) group. The levels of IL-6 and IL-10 were also elevated, especially after LPS administration. Treatment with mianserin resulted in a significant lowering of TNFα and IL-6 levels both in LPS-treated and LPS-untreated animals. There was also a decrease in IL-10 concentration in LPS-treated stressed animals. CONCLUSIONS: The results confirm an increase in proinflammatory cytokines in the blood of rats with experimentally induced depression and show the protective role of the activity of mianserin on the cytokine levels, expressed in a lowering of TNFα and IL-6 levels in stressed animals, and of IL-10 levels after LPS administration.

Rosmarinus officinalis L. leaf extract improves memory impairment and affects acetylcholinesterase and butyrylcholinesterase activities in rat brain.

Rosmarinus officinalis L. leaf as part of a diet and medication can be a valuable proposal for the prevention and treatment of dementia. The aim of the study was to assess the effects of subchronic (28-fold) administration of a plant extract (RE) (200 mg/kg, p.o.) on behavioral and cognitive responses of rats linked with acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity and their mRNA expression level in the hippocampus and frontal cortex. The passive avoidance test results showed that RE improved long-term memory in scopolamine-induced rats. The extract inhibited the AChE activity and showed a stimulatory effect on BuChE in both parts of rat brain. Moreover, RE produced a lower mRNA BuChE expression in the cortex and simultaneously an increase in the hippocampus. The study suggests that RE led to improved long-term memory in rats, which can be partially explained by its inhibition of AChE activity in rat brain.

Ethanol affects acylated and total ghrelin levels in peripheral blood of alcohol-dependent rats.

There is a hypothesis that ghrelin could take part in the central effects of alcohol as well as function as a peripheral indicator of the changes which occur during long-term alcohol consumption. The aim of this study was to determine a correlation between alcohol concentration and acylated and total form of ghrelin after a single administration of alcohol (intraperitoneal, i.p.) (experiment 1) and prolonged ethanol consumption (experiment 2). The study was performed using Wistar alcohol preferring (PR) and non-preferring (NP) rats and rats from inbred line (Warsaw High Preferring, WHP; Warsaw Low Preferring, WLP). It was found that ghrelin in ethanol-naive WHP animals showed a significantly lower level when compared with the ethanol-naive WLP or Wistar rats. After acute ethanol administration in doses of 1.0; 2.0 and 4.0 g/kg, i.p., the simple (WHP) or inverse (WLP and Wistar) relationship between alcohol concentration and both form of ghrelin levels in plasma were found. Chronic alcohol intake in all groups of rats led to decrease of acylated ghrelin concentration. PR and WHP rats, after chronic alcohol drinking, had lower levels of both form of ghrelin in comparison with NP and WLP rats, respectively, and the observed differences in ghrelin levels were in inverse relationship with their alcohol intake. In conclusion, it is suggested that there is a strong relationship between alcohol administration or intake, ethanol concentration in blood and both active and total ghrelin level in the experimental animals, and that ghrelin plasma concentration can be a marker of alcohol drinking predisposition.

Reactions of N(3) -substituted amidrazones with cis-1,2-cyclohexanedicarboxylic anhydride and biological activities of the products.

A series of novel compounds were synthesized in reactions of N(3) -substituted amidrazones with cis-1,2-cyclohexanedicarboxylic anhydride: linear, isoindole, and triazole derivatives. All new structures were confirmed by H(1) NMR and IR spectrometry as well as elemental analysis. Potential biological effects of new compounds were predicted with the Prediction of Activity Spectra for Substances (PASS) program. Antiviral, antibacterial, analgesic, and anti-inflammatory activities were experimentally verified.

The influence of a standardized soybean extract (Glycine max) on the expression level of cytochrome P450 genes in vivo.

OBJECTIVE: Soybean isoflavones are phytoestrogens that reduce menopausal symptoms and decrease the risk of certain chronic diseases, such as cancer and cardiovascular diseases. Despite the widespread use of soybean isoflavones as functional food and dietary supplements, data regarding the safety as well as herb-drug interactions, remain scarce. The aim of the study was to investigate the influence of soybean extract on the expression levels of cytochrome P450 (CYP) genes using real-time PCR (RT-PCR). MATERIALS AND METHODS: Male Wistar rats were fed a standardized soybean extract containing 37% isoflavones (100 mg/kg) for 3 and 10 days. cDNA was synthesized from total RNA isolated from the liver using reverse transcription. The level of CYP genes expression was analyzed using RT-PCR method. RESULTS: Soybean extract administration resulted in a significant increase of CYP1A1 expression level compared with the control group (1.5-fold; p < 0.05). An inductory effect was also observed for CYP2D1 by 32% (p < 0.01) after 10 days of treatment. No statistically significant differences were noted for CYPIA2, CYP2C6 and CYP3A2. In case of CYP3A1, the mRNA level of this gene was reduced by almost 35% (p < 0.05) both, after 3 and 10 days. CYP2D2 expression was also inhibited by the extract, but to a lesser degree when compared to CYP3A1. Moreover insignificant decrease of CYP2E1 expression level by 25% (p < 0.01) was observed after 3 days of treatment. CONCLUSIONS: These findings suggest that soybean extract may change the expression of CYP enzymes involved in biotransformation of xenobiotics (drugs, procarcinogens).

[Influence of standardized extract of Epilobium angustifolium on estrogen receptor alpha and beta expression in in vivo model]. / Wplyw standaryzowanego wyciqgu z Epilobium angustifolium na ekspresj mRNA receptorów estrogenowych alpha i beta w modelu in vivo.

OBJECTIVE: Evaluation of the influence of the standardized extract from the herb of Epilobium angustifolium on ERalpha and ERbeta mRNA expression in rat ventral prostate tissue and free serum estradiol level. MATERIALS AND METHODS: Rats were divided into 6 groups with 10 animals. ERalpha and ERbeta mRNA expression in rat ventral prostate tissue level was performed using real-time PCR method in Light Cycler system. Serum-free estradiol was evaluated using immunoenzymatic technique. RESULTS: In our experimental model there was an increase of ERa mRNA level by 9% and decrease by 36% of ERbeta mRNA level in ventral prostate tissue in rats administrated with testosterone and E. angustifolium extract, in comparison with testosterone alone administrated animals. CONCLUSIONS: E. angustifolium standardized extract influenced the expression of estrogen receptor alpha and beta mRNAs in differential manner which may suggest its potentially therapeutic properties or causing of adverse effects in pharmacotherapy of estrogen-related disorders. More complex studies should be undertaken to evaluate safety and to improve the efficacy of using this herbal extract.

[The influence of a standardized soybean extract (Glycine max) on the expression level of CYP3A enzymes and pregnane X receptor in in vivo model]. / Wplyw standaryzowanego ekstraktu sojoweg (Glycine max) na poziom ekspresji enzymów CYP3A i receptora pregnanu X w modelu in vivo.

OBJECTIVE: Soybean phytoestrogens, such as genistein and daidzein, have become a popular alternative for women undergoing the treatment of menopause symptoms. These isoflavones are also commonly used in traditional medicine in the prevention and treatment of osteoporosis, cardiovascular diseases and cancer Despite the widespread use of soybean preparations as functional foods and dietary supplements, data regarding the safety as well as interactions between herbal medicines and synthetic drugs, especially with antineoplastic agents, remain scarce. Therefore, the aim of the present study was to determine the influence of soybean extract on the expression levels of CYP3A and PXR genes using real-time PCR (RT-PCR). MATERIALS AND METHODS: Male Wistar rats were given a standardized soybean extract (100 mg/kg p.o.) for 3 and 10 days. Total RNA isolated from the liver tissue was transcribed into cDNA. The level of CYP3A 1/2 and PXR mRNAs expression was analyzed by real-time quantitative PCR using SYBR Green I dye. RESULTS: Our findings showed that soybean extract containing 37% isoflavones resulted in a significant decrease of CYP3A1 expression level by almost 35% (p<0.05), both after 3 and 10 days, when compared with the control group. No statistically significant differences were noted for CYP3A2 enzyme and the PXR nuclear factor. CONCLUSIONS: These results suggest that soybean extract can decrease the CYP3A1 (homolog to human CYP3A4) expression and may participate in clinically significant interactions with drugs metabolized by CYP3A4 enzyme. Moreover it is postulated that gene expression of CYP3A1 and CYP3A2 (homolog to human CYP3A5) can be regulated indirectly by the PXR transcription factor.

IL-12 serum levels in patients with type 2 diabetes treated with sulphonylureas.

Interleukin-12 (IL-12) has been identified as a pro-inflammatory cytokine which is thought to contribute to the development of atherosclerosis. However, to date, the various associations between factors related to the course of type 2 diabetes, like metabolic compensation, beta cell secretory dysfunction, insulin resistance and IL-12 serum levels, remain unclear. Our study involved 41 patients with type 2 diabetes, 19 patients with coronary artery disease (CAD), and 19 healthy controls. We measured serum levels of fasting glucose, HbA(1)c, 1,5-anhydro-d-glucitol, and lipids. In addition, serum levels of C-peptide, insulin, proinsulin and IL-12 were assayed. HOMA(IR) score was calculated. The serum concentrations of IL-12 were higher in diabetics than in either patients with CAD or healthy controls, and were correlated with BMI, C-peptide, insulin, HOMA(IR), proinsulin and HDL serum levels. Multiple regression analysis revealed that the IL-12 serum level in type 2 diabetics primarily is dependent upon fasting proinsulin concentration. Our results demonstrate that elevated IL-12 serum levels in type 2 diabetics treated with sulphonylureas are induced especially by peripheral insulin resistance and beta cells dysfunction, as expressed by fasting serum proinsulin levels. This finding gives us hope that treatment to decrease peripheral insulin resistance and to avoid excessive proinsulin secretion might be successful in the prevention of IL-12-induced atherosclerosis.

Pro-atherogenic alterations in T-lymphocyte subpopulations related to acute hyperglycaemia in type 2 diabetic patients.

BACKGROUND: T cells are among the earliest cells to infiltrate the arterial intima during the initial stages of atherosclerosis. Alterations in the peripheral blood lymphocyte distribution might be associated with intensive lymphocytes extravasation and stimulation of atherosclerotic plaque development. Epidemiological data reveal that short-term postprandial hyperglycemia is a significant risk factor for coronary heart disease. Using a parameter that indicates recently-past acute hyperglycemia, 1,5-anhydro-D-glucitol (1,5-AG), the aim of the present study was to elucidate which alterations in peripheral blood T-lymphocytes, if any, are associated with acute hyperglycemia in patients with type 2 diabetes mellitus (DM) and, thus, might be involved in the progression of atherosclerosis. METHODS AND RESULTS: Measurement of fasting glucose level, glycated hemoglobin A(1c), 1,5-AG, lipid profile and lymphocyte receptors expression (CD3+, CD4+, CD8+, CD8+28+, CD+28 -) was performed in 97 patients with type 2 DM, 23 patients with coronary heart disease, and 15 healthy controls. The mean CD3+, CD4+, CD8+28 - and CD8+28+ lymphocyte counts were significantly higher in the DM patients than in both control groups. Multiple regression analysis revealed that CD4+ and CD8+28- lymphocyte counts primarily were dependent on 1,5-anhydro-D-glucitol plasma levels. CONCLUSIONS: These results suggest that acute hyperglycemia results in the progression of atherosclerosis in type 2 DM, at least in part through changes in CD4+ and CD8+28- lymphocyte subsets.

Hypoglycaemic effect of quinolizidine alkaloids--lupanine and 2-thionosparteine on non-diabetic and streptozotocin-induced diabetic rats.

The hypoglycaemic effects of two quinolizidine alkaloids: lupanine and 2-thionosparteine were examined in non-diabetic and in streptozotocin-induced diabetic rats. The model of experimental diabetes can be considered to be related to diabetes mellitus type 2 with regards to the impairment of beta-cells' secretory function. A single intraperitoneal injection of 2-thionosparteine at a dose of 8.6 mg/kg lowered the blood glucose levels in diabetic rats at 90 and 120 min after administration and showed similar hypoglycaemic effects to glibenclamide and sparteine, which were used as reference substances. In contrast to glibenclamide, 2-thionosparteine did not result in a significant increase in plasma insulin levels in diabetic rats; an increase was only observed in the non-diabetic group. It was found that lupanine did not exert hypoglycaemic potency in diabetic and in non-diabetic animals and did not significantly increase plasma insulin concentration independent of the group examined. From this study we can state that 2-thionosparteine, but not lupanine, is confirmed to be a possible plasma glucose lowering agent. It is possible that 2-thionosparteine-dependent decrease in blood glucose level is not the only result of this drug's related insulin secretion.

[Folic acid: new indications for an old well-known drug]. / Kwas foliowy--nowe wskazania dla dawno znanego leku.

UNLABELLED: This review deals with the well known drug--folic acid, for which new indications have been discovered in recent years. The drug was used for 50 years in anemia megaloblastia, other anemias with bone marrow hypofunction, in leucopenia and granulocytopenia, in celiac disease and sprue and folic acid deficit during oral anticonceptive drug use. From recent investigations follows that there are strong indications for the use of folic acid in a periconceptual period and during pregnancy (at least 600 micrograms per day) and during lactation--to prevent neural tube defects. To prevent orofacial cleft defects a dose 5 mg a day mast be used. Another important new indication is hyperhomocysteinemia. Folic acid, B12 and B6 vitamins are involved in the metabolic removal of homocysteine, but folic acid deficit occurs the most often. The results of hyperhomocysteinemia are increased risk of arteriosclerosis coronary heart diseases and stroke. RESULTS: Folic acid is an important factor in preventing congenital defects (especially of the neural tube) and for lowering the risk of coronary heart disease and stroke due to hyperhomocysteinemia (together with vitamins B6 and B12).

[Pharmacological justification for the use of new antidepressant drugs]. / Farmakologiczne uzasadnienie skutecznosci stosowania nowych leków przeciwdepresyjnych.

This review compares a new generation of antidepressants with standard tricyclic antidepressants. It is now believed that an ideal antidepressant should be characterised by high therapeutic efficacy, good tolerance, safety of use related to low incidence of adverse effects, little potential for interactions and low cost of use. Mirtazapine, nefazodone and venlafaxine have therapeutic efficacy which is comparable with that of tricyclic antidepressants and their lack of (or insignificant) receptor effect as well as double mechanism of action makes them a safer therapeutic option.

Effect of multiple ifenprodil or spermidine treatment on social recognition in rats.

We investigated the effects of multiple (21 x) ifenprodil (1.0 mg/kg, i.p.)-[IF] and spermidine (5.0 mg/kg, i.p.)-[SP] administration on short-term memory using the social recognition test in rats. The influence of a single (lx) injection of IF and SP was also established. 1x IF or SP treatment showed a statistically insignificant tendency to impair social memory task. In contrast, 21 x SP treatment facilitated short-term memory when compared with 1x SP administration. 21x IF had no affect on the memory paradigm. The results of the present study indicate that the prolonged systemic treatment of IF or SP in relatively low doses causes no impairment of short-term memory in rats.

Role of neuropeptides in antidepressant and memory improving effects of venlafaxine.

The aim of this study has been to investigate the effects of vasopressin and oxytocin on antidepressive and memory improving effects of venlafaxine. Male Wistar rats weighing 180-200 g were used in the study. Venlafaxine (20 mg/kg) was administered po 30 min before the test once, and for 7 and 14 days in the chronic experiments. Oxytocin (1 microg/kg) ip and vasopressin (1 microg/kg) sc were administered only once on the test day, 60 min before the tests. The animals were subjected to Porsolt's test for testing antidepressant activity, and their memory functions (working and spatial memory) were evaluated in the maze test and Morris Water Maze test. Antidepressant effects of venlafaxine could be observed already after single drug administration and the effect was maintained during 7 days of drug administration. Oxytocin also exhibited antidepressant activity, and concurrent administration of venlafaxine and oxytocin helped to maintain antidepressant activity of venlafaxine. Vasopressin was devoid of antidepressant action, yet concurrent administration of vasopressin and venlafaxine did not suppress antidepressant activity of the latter. In the chronic experiment, there was no shortening of passive swimming time. Venlafaxine improved memory in the labyrinth test and in the spatial memory test, whereas oxytocin did not affect memory of the tested animals. Joint administration of venlafaxine and oxytocin did not produce memory improving effect observed after administration of venlafaxine only. Vasopressin improved memory and joint administration of venlafaxine and vasopressin maintained the memory improving effect induced by vasopressin. The regulatory role of neuropeptides and new antidepressant drugs, e.g. venlafaxine in mood status and memory functions may depend on the interactions between monoaminergic and neuropeptidergic systems.