Oral <Emphasis Type="Italic">L</Emphasis>-Carnitine Supplementation and Exercise Metabolism

Summary. Oral L-carnitine supplementation is frequently reported to have beneficial effects on exercise capacity in clinical populations and has been considered as a potential ergogenic aid for endurance athletes. However, this latter view is largely unsubstantiated possibly due to many experimental studies being poorly controlled or difficult to compare. The potential for oral L-carnitine supplementation to influence skeletal muscle carnitine content has been questioned and there are several key factors identified that may explain variations between study outcomes. Recent more well controlled research suggests some potential for L-carnitine to act as a key regulator of cellular stress, possibly through an impact on the integration of carbohydrate and lipid metabolism, and this work should be followed up in future by well controlled studies in both athlete and clinical subject groups.     

Effects of <Emphasis Type="Italic">L</Emphasis>-Carnitine Supplementation in Sows

Summary. In recent years L-carnitine has been used increasingly in animals. This review gives an overview of the effects of dietary L-carnitine supplementation during pregnancy and lactation on the reproductive performance of sows. In one investigation L-carnitine supplementation during pregnancy increased the number of piglets born to sows. Other studies showed heavier litters in sows supplemented with L-carnitine compared with control sows, and litters of L-carnitine supplemented sows gained more weight during the suckling period than litters of control sows. This effect might be due to more vigorous suckling by piglets of L-carnitine supplemented sows, causing the sows’ milk production to rise. At negative energy balance during lactation L-carnitine supplemented sows are able to mobilize more energy from adipose tissue, which can be used for the production of surplus milk. In conclusion, recent studies clearly show that dietary L-carnitine supplementation increases the reproductive performance of sows. This finding suggests that the amount of L-carnitine synthesized endogenously does not cover the requirement for maximum sow performance during pregnancy and lactation.     

Current Methods for Determination of <Emphasis Type="Italic">L</Emphasis>-Carnitine and Acylcarnitines

Summary. L-Carnitine as endogenous compound plays an important role within several metabolic pathways and a deficiency of L-carnitine can cause adverse effects in physiological and/or mental state of health and disease. The prevention of diseases related to carnitine deficiency requires, first of all, the exact determination of L-carnitine and its esters in biological material at pmol/cm³ level. A series of analytical procedures based on biochemical assays as well as on physical methods are available today. Determination of free and total carnitine is sometimes sufficient for a clinical diagnosis, but in most cases, such as in newborn screening for genetic disorders, detailed qualitative and quantitative L-carnitine/acylcarnitine profiling is needed. Technological progress has also revolutionized the determination of carnitines. Today, comprehensive and diagnostically relevant information can be obtained by mass spectrometry. An overview is given of the technical and methodological developments in carnitine analysis and some applications, such as in neonatal screening, diabetes mellitus, and cardiomyopathy.     

Large Scale Bioprocess for the Production of Optically Pure <Emphasis Type="Italic">L</Emphasis>-Carnitine

Summary. A competitive production method using the biotransformation of 4-butyrobetaine to enantiomerically pure L-carnitine was developed and scaled-up by Lonza. The process produces L-carnitine in 99.5% yield, and >99.9% enantiomeric excess (ee). Continuous and discontinuous processes were developed but the fed-batch process was found to be economically the most favourable process mode.     

Effects of <Emphasis Type="Italic">L</Emphasis>-Carnitine on Sucrose-Induced Hyperlipidaemia

Summary. L-Carnitine, L-(−)-β-hydroxy-γ-trimethylaminobutyrate, plays an important role as a factor necessary for the transport of long-chain fatty acids into the mitochondria. In order to investigate the influence of L-carnitine on hyperlipidaemias, the experimental model of the sucrose-induced hypertriglyceridaemia of the rat was used. In these experiments L-carnitine in the dose of 11 mg per day and 100 g body weight (over the period of 1 week) was able to antagonize the sucrose-induced hypertriglyceridaemia and the increase of serum free fatty acid level in female rats of the Wistar strain. Carnitine administration did not change the activities of lipogenic enzymes and fatty acid synthesis in the liver. However, L-carnitine increases the rate of hepatic fatty acid oxidation. Our results indicate a hypotriglyceridemic and free fatty acid lowering effect of L-carnitine, and suggest the use of this compound in the therapy of hyperlipidaemias.     

Protective Effects of <Emphasis Type="Italic">L</Emphasis>-Carnitine on Myocardium of Experimentally Diabetic Rats with Additional Ischaemia and Reperfusion

Summary. We investigated the protective effects of L-carnitine against damage to the heart caused by diabetes-induced alterations and additional ischaemia and reperfusion in diabetic BB/OK rats using histological techniques, morphometry, biochemical parameters of oxidative stress, and SOD expression. The results revealed that diabetes-induced morphological changes were partly improved or nearly prevented by substitution of L-carnitine, which also seemed to improve the reduced tolerance of diabetic myocardium towards ischaemia/reperfusion with respect to morphological parameters. Immunohistochemical and biochemical parameters of oxidative stress such as SOD protein expression as well as SOD and GPx activity indicate increased free oxygen radical level in the ischaemic/reperfused diabetic myocardium, which is clearly decreased by L-carnitine treatment. We suggest that L-carnitine may be an adequate “causal” agent in the protection of myocardial alterations in diabetes with additional ischaemia and reperfusion, as it stabilizes mitochondrial and cellular function and acts through its antioxidative or radical scavenging potential. Further investigations are necessary to determine an approach towards adjuvant treatment of diabetic myocardial complications using L-carnitine.     

Synthesis of 6-methyl-8<Emphasis Type="Italic">H</Emphasis>-dibenzo[<Emphasis Type="Italic">a</Emphasis>,<Emphasis Type="Italic">g</Emphasis>]quinolizin-8-imines <Emphasis Type="Italic">via Reissert</Emphasis> compounds

Alkylation of Reissert compounds derived from 3-methylisoquinolines with several 2-cyanobenzylbromides followed by hydrolytic cleavage provided the corresponding 1-benzyl-3-methylisoquinolines. Treatment of the latter with methylmagnesiumiodide caused cyclization to the title compounds rather than formation of 2-acetylbenzylisoquinolines.     

Rapid molecular typing of <Emphasis Type="Italic">Tuber melanosporum</Emphasis>, <Emphasis Type="Italic">T. brumale</Emphasis> and <Emphasis Type="Italic">T. indicum</Emphasis> from tree seedlings and canned truffles

We have developed new DNA extraction and purification procedures for investigation of mycorrhized seedlings and canned truffles. Use of these procedures on approximately 100 mg initial material enabled good sample representation. For mycorrhized seedlings, Taq polymerase inhibitors were discarded irrespective of tree species. In routine analysis we systematically used consensus primers ITS1/ITS4 to check the absence of Taq polymerase inhibitors and the presence of fungus DNA. Positive response with ITS validates other positive or negative PCR results. Absence of amplification with ITS prevents validation of other results. For canned truffles, DNA harvested from ascocarps sterilized for one and a half hours at 115°C was amplified with specific primers. We have developed consensus primers, named R12/F12, to check for the presence of amplifiable fungus DNA and the absence of Taq polymerase inhibitors. Here also, positive response with consensus R12/F12 validates other positive or negative PCR results. We have developed one primer pair specific for T. brumale and another specific for T. melanosporum. We can then characterize these two taxa, which enables the use of truffle or truffled French designations. We can also characterize T. indicum, the Asiatic black truffle that might fraudulently be sold as T. melanosporum and T. brumale. These three specific primer pairs were used independently of DNA extraction from tree seedlings or canned truffles. Our process is specific, sensitive, convenient, and quick.J.P. Douet and D. Mabru have contributed equally to this work     

Antioxidant flavonoids from <Emphasis Type="Italic">Tamus communis</Emphasis> ssp. <Emphasis Type="Italic">cretica</Emphasis>

A new flavonoid, kaempferol-3,4′-di-O-α-L-rhamnopyranoside (1), and three known flavonoids (2–4) were isolated from the aerial parts of T. communis L. The structure of the new compound was elucidated on the basis of spectroscopic data. Compounds 1 and 2 showed significant antioxidant activity (IC₅₀ 187.151 ± 0.821 μM, and 92.079±0.513 μM, respectively), whereas compounds 3 and 4 showed moderate activity in DPPH free radical scavenging assays. Published in Khimiya Prirodnykh Soedinenii, No. 3, pp. 295–297, May–June, 2009.     

Lipids from <Emphasis Type="Italic">Halostachys caspica</Emphasis> and <Emphasis Type="Italic">Halocharis hispida</Emphasis>

The composition of lipids from the aerial parts of two species of halophytes from the family Chenopodiaceae, Halostachys caspica C. A. Mey. and Halocharis hispida Bge. was determined. Neutral lipids (NL, 62.1 and 54.2%, respectively) dominated the total lipids (TL) of these plants. More than a third of the NL were esters of aliphatic alcohols and phytosterols (FAE). Fatty acids 16:0, 18:1, and 18:2 dominated the acids of FAE; 16:0, 18:1, and 18:3, the phospholipids. The principal fatty acids of glycolipids were unsaturated acids (68.3 and 75.1%) with linolenic acid dominating (44.9 and 43.5%). Presented at the 7th International Symposium on the Chemistry of Natural Compounds, Tashkent, October 16–18, 2007. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 276–278, May–June, 2009.     

Screening and Immobilization <Emphasis Type="Italic">Burkholderia</Emphasis> sp. GXU56 Lipase for Enantioselective Resolution of (<Emphasis Type="Italic">R</Emphasis>,<Emphasis Type="Italic">S</Emphasis>)-Methyl Mandelate

Microorganisms producing lipase were isolated from soil and sewage samples and screened for enantioselective resolution of (R,S)-methyl mandelate to (R)-mandelic acid. A strain designated as GXU56 was obtained and identified as Burkholderia sp. Preparing immobilized GXU56 lipase by simple adsorption on octyl sepharose CL-4B, the optimum temperature was shifted from 40 °C (free lipase) to 50 °C (immobilized lipase), and the optimum pH was shifted from 8.0 (free lipase) to 7.2 (immobilized lipase). The immobilized enzyme displayed excellent stability in the pH range of 5.0–8.0, at the temperatures below 50 °C and in organic solvents compared with free enzyme. Enantioselectivity ratio for (R)-mandelic acid (E) was dramatically improved from 29.2 to more than 300 by applying immobilized lipase in the resolution of (R,S)-methyl mandelate. After five cycles of use of immobilized lipase, conversion and enantiomeric excess of (R)-mandelic acid were 34.5% and 98.5%, respectively, with enantioselectivity ratio for (R)-mandelic acid (E) of 230. Thus, octyl-sepharose-immobilized GXU56 lipase can be used as a bio-resolution reagent for producing (R)-mandelic acid.     

Stress Response Kinetics of Two Nisin Producer Strains of <Emphasis Type="Italic">Lactococcus lactis</Emphasis> spp. <Emphasis Type="Italic">lactis</Emphasis>

The purpose of this study is to determine the survival and nisin production behaviors of two strains of Lactococcus lactis under different stress conditions that represent the food ecosystem. In this respect, the survival ratios of two nisin producers were determined under different pH, temperature, NaCl, and bile salt concentrations. Then, nisin production levels of the strains were determined at each stress conditions. Both strains had similar growth or inactivation patterns under the same stress conditions. NaCl and bile salt stresses on the survival ratio of the strains could be successfully described by the exponential decay function, whereas Gaussian function produced good fits for temperature and pH stresses. The nisin activity of two nisin producers (in their mid-exponential and/or early stationary phase) decreased dramatically under all stress conditions, except osmotic (NaCl) and low temperature applications. The results of this study showed that two nisin producers had similar adaptive responses under severe stress conditions, which could be described by appropriate mathematical equations. Moreover, the effect of harsh environment on the nisin activity of L. lactis strains depends on the stress factors applied.     

An efficient <Emphasis Type="Italic">Mannich</Emphasis>-type synthesis of <Emphasis Type="Italic">bis</Emphasis>(pyrido[2,1-<Emphasis Type="Italic">b</Emphasis>][1,3,5]thiadiazin-7-yl)-methanes

Morpholinium 3-cyano-4-methyl-6-oxo-1,6-dihydropyridine-2-thiolate upon treatment with primary amines and a formaldehyde excess under mild conditions produces bis(pyrido[2,1-b][1,3,5]thiadiazin-7-yl)methane derivatives in good yields (67–87%). Correspondence: Victor V. Dotsenko, State Enterprise “Luganskstandartmetrology”, 91021 Lugansk, Ukraine.     

Syntheses of (<Emphasis Type="Italic">E</Emphasis>)- and (<Emphasis Type="Italic">Z</Emphasis>)-3-styrylchromones

Several (E)- and (Z)-3-styrylchromones were prepared by two different methodologies, the Wittig reaction of chromone-3-carboxaldehyde with benzylic ylides and the Knoevenagel condensation of chromone-3-carboxaldehyde with phenylacetic acids in the presence of potassium tert-butoxide under microwave irradiation. The Knoevenagel reaction followed by a decarboxylation offered an efficient and diastereoselective method for preparing (E)-3-styrylchromones in a shorter reaction time. It was also demonstrated that phenylacetic acid can also be substituted with success by phenylmalonic acid. The stereochemistry of all products was assigned by NMR experiments. Correspondence: Artur M. S. Silva, Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal.     

New flavonoid-<Emphasis Type="Italic">C</Emphasis>-glycosides from <Emphasis Type="Italic">Triticum aestivum</Emphasis>

Two new flavonoid-C-glycosides named triticuside A (1a) and triticuside B (1b) were isolated from bran of Triticum aestivum L. The structures of the two new compounds were elucidated by spectral techniques including ¹H NMR, ¹³C NMR as well as HSQC, HMBC, and COSY. Published in Khimiya Prirodnykh Soedinenii, No. 2, pp. 135–137, March–April, 2008.     

Splitting of the <Emphasis Type="Italic">d</Emphasis><Superscript><Emphasis Type="Italic">N</Emphasis></Superscript> atomic states in icosahedral 3<Emphasis Type="Italic">d</Emphasis> metal endofullerenes M@C<Subscript>60</Subscript>

Group-theoretical and quantum-chemical investigations of the spectrum of low-lying excited states have been performed by the ROHF and FCI-RAS (Full CI in Restricted Active Space) methods for 3d metal endofullerenes (MEFs) M@C₆₀ (M =Mn, Cr, and Fe) in different charged states. The major purpose of this study is quantum-chemical verification of the anomalous (“non-Bethe’s”) character of splitting of the d ^N atomic states in an electrostatic field with icosahedral symmetry, predicted previously within the theory of integral invariants theory. The interrelation between the integral invariants theory and the quantumchemical methods applied in this work is considered in detail. Our calculations suggest that the d ^N atomic states in the icosahedral field generated by fullerene C60 (I _h ) on a metal atom (ion) remain non-split for different charged states of the metal and C₆₀. Reasons for this phenomenon and other possible approaches to verification of the prediction are discussed. It is demonstrated that the d ^N states of the encapsulated metal are split in icosahedral 3d MEFs only under very strong compression of these structures.     

A new stilbene glycoside from the <Emphasis Type="Italic">n</Emphasis>-butanol fraction of <Emphasis Type="Italic">Veratrum dahuricum</Emphasis>

A new stilbene glycoside, 5-methylresveratrol-3,4′-O-β-D-diglucopyranoside (1), was isolated from the n-butanol fraction of the rhizomes of Veratrum dahuricum, together with five known stilbenoids: resveratrol-3-O-β-D-glycoside (2), 4′-methylresveratrol-3-O-β-D-glycoside (3), oxyresveratrol-4′-O-β-D-glycoside (4), oxyresveratrol-3-O-β-D-glycoside (5), and oxyresveratrol-3,4′-O-β-D-diglycoside (6), and found for the first time in the investigated plant. The structures of six isolates were identified on the basis of 1D and 2D NMR data. Compounds 1–6 showed platelet aggregation inhibition, and compound 1 had an IC₅₀ value of 383.6 μM against platelet aggregation induced by AA. Published in Khimiya Prirodnykh Soedinenii, No. 3, pp. 279–282, May–June, 2009.     

Triterpene glycosides from <Emphasis Type="Italic">Lonicera</Emphasis>. Isolation and structural determination of seven glycosides from flower buds of <Emphasis Type="Italic">Lonicera macranthoides</Emphasis>

The structures of seven triterpene glycosides (1–7), of which the 23-O-acetyl, 28-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranosyl ester of hederagenin 3-O-β-D-glucopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→2)-O-α-L-arabinopyranoside (2) was new, from the flower buds of Lonicera macranthoides were established using chemical and NMR spectroscopic methods. Published in Khimiya Prirodnykh Soedinenii, No. 1, pp. 32–34, January–February, 2008.     

Expression Analysis and Characteristics of <Emphasis Type="Italic">Profilin</Emphasis> Gene from Silkworm, <Emphasis Type="Italic">Bombyx mori</Emphasis>

A recombinant Bombyx mori profilin protein (rBmPFN) was overexpressed in Escherichia coli BL21. Purified rBmPFN was used to generate anti-BmPFN polyclonal antibody, which were used to determine the subcellular localization of BmPFN. Immunostaining indicated that profilin can be found in both the nucleus and cytoplasm but is primarily located in the cytoplasm. Real-time RT-PCR and Western blot analyses indicated that, during the larvae stage, profilin expression levels are highest in the silk gland, followed by the gonad, and are lowest in the fatty body. Additionally, BmPFN expression begins during the egg stage, increases during the larvae stage, reaches a peak during the pupa stage, and decreases significantly in the moth. Therefore, we propose that BmPFN may play an important role during larva stage development, especially in the silk gland.     

Glucofructans from <Emphasis Type="Italic">Taraxacum officinale</Emphasis> roots

The structure of glucofructans from Taraxacum officinale roots growing in Buryatia was studied by chemical, chromatographic, and spectral methods. It was found that fructose,glucose, saccharose, 1-kestose, and nystose were present in the free state. The structures of the two dominant polymeric compounds, TGf-1 (5.7 kDa) and TGf-2 (2.6 kDa), which were linear inulin-type macromolecules consisting of fructofuranose units bonded through β-(2→1)-bonds, were studied. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 125–127, March–April, 2009.