Determination of the conversion factor for silymarin - a solution for the problems associated with new assay methods in the pharmacopoeia.

In pharmacopoeial monographs for herbal drugs and herbal preparations, conventional assay methods such as colorimetry or spectophotometric assays are often replaced by modern, more specific and reliable methods, e.g. liquid chromatography (LC). However, existing dosage recommendations in the monographs on efficacy and safety of herbal medicinal products which are an important basis for licensing procedures do not refer to the mandatory new methods but to the existing photometric methods. The laboratory comparison of the determination of silymarin of Milk Thistle extract shows that a conversion factor can be calculated which allows a correlation between the new and the existing method. It is suggested that this factor should be included in the Ph. Eur. monograph on Milk Thistle extract, allowing reference to dosages given in official monographs (e.g. ESCOP, HMPC). The solution to use a conversion factor should also be applied to other herbal drugs and herbal preparations, especially for standardised extracts.

Anthocyanin- and proanthocyanidin-rich extracts of berries in food supplements--analysis with problems.

The fundamental nutritional benefit of fruit and vegetables in the prevention of degenerative diseases--especially in the light of the current "anti-aging wave"--has directed the attention of scientists and consumers to a variety of berry fruits and their constituents. Many of these fruits, e.g. blueberries, elderberries or cranberries, have a long tradition in European and North American folk medicine. Based on these experiences and due to the growing interest the number of food supplements on the market containing fruit powders, juice concentrates or extracts of these fruits has increased considerably. Advertising for these products mainly focusses on the phenolic compounds, especially the anthocyanins and proanthocyanidins and their preventive effects. Most of the preparations are combinations, e.g. of extracts of different fruits with vitamins and trace elements, etc. which are labelled in a way which does not allow a comparison of the products. Typically, information on the extraction solvent, the drug: extract ratio and the content of anthocyanins and proanthocyanidins is missing. Besides that, the analysis of these polyphenols causes additional problems. Whereas the quality control of herbal medicinal products is regulated in detail, no uniform requirements for food supplements are existing. A broad spectrum of methods is used for the assay of the constituents, leading to differing, incomparable results. In addition to that, the methods are quite interference-prone and consequently lead to over- or underestimation of the contents. This publication provides an overview of some selected berries (lingonberry, cranberry, black elderberry, black chokeberry, black currant, blueberry), their constituents and use. The analytical methods currently used for the identification and quantification of the polyphenols in these berries are described, including an evaluation of their advantages and disadvantages.

Self-monitoring of blood glucose in type 2 diabetes and long-term outcome: an epidemiological cohort study.

AIMS/HYPOTHESIS: The aim of this study was to obtain epidemiological data on self-monitoring of blood glucose (SMBG) in type 2 diabetes and to investigate the relationship of SMBG with disease-related morbidity and mortality. METHODS: The German multicentre Retrolective Study 'Self-monitoring of Blood Glucose and Outcome in Patients with Type 2 Diabetes' (ROSSO) followed 3,268 patients from diagnosis of type 2 diabetes between 1995 and 1999 until the end of 2003. Endpoints were diabetes-related morbidity (non-fatal myocardial infarction, stroke, foot amputation, blindness or haemodialysis) and all-cause mortality. SMBG was defined as self-measurement of blood glucose for at least 1 year. RESULTS: During a mean follow-up period of 6.5 years, 1,479 patients (45.3%) began SMBG prior to an endpoint and an additional 64 patients started SMBG after a non-fatal endpoint. Interestingly, many patients used SMBG while being treated with diet or oral hypoglycaemic drugs (808 of 2,515, 32%). At baseline, the SMBG cohort had higher mean fasting blood glucose levels than the non-SMBG cohort (p<0.001), suggesting that insufficient metabolic control was one reason for initiating SMBG. This was associated with a higher rate of microvascular endpoints. However, the total rate of non-fatal events, micro- and macrovascular, was lower in the SMBG group than in the non-SMBG group (7.2 vs 10.4%, p=0.002). A similar difference was found for the rate of fatal events (2.7 vs 4.6%, p=0.004). Cox regression analysis identified SMBG as an independent predictor of morbidity and mortality, with adjusted hazard ratios of 0.68 (95% CI 0.51-0.91, p=0.009) and 0.49 (95% CI 0.31-0.78, p=0.003), respectively. A better outcome for both endpoints was also observed in the SMBG cohort when only those patients who were not receiving insulin were analysed. CONCLUSIONS/INTERPRETATION: SMBG was associated with decreased diabetes-related morbidity and all-cause mortality in type 2 diabetes, and this association remained in a subgroup of patients who were not receiving insulin therapy. SMBG may be associated with a healthier lifestyle and/or better disease management.

Vitamin D inadequacy among post-menopausal women: a systematic review.

BACKGROUND: Vitamin D inadequacy has been studied extensively, due to concerns about ageing populations, associations with osteoporosis and other disorders (including non-musculoskeletal), and high prevalence. AIM: To review recent reports on the prevalence of vitamin D inadequacy among post-menopausal women with and without osteoporosis and/or other musculoskeletal diseases. DESIGN: Systematic review. METHODS: We reviewed publications in the past 10 years reporting prevalence estimates for vitamin D inadequacy, reported as serum 25(OH)D values below various levels. Thirty published studies in the English language were identified, from January 1994 through April 2004. RESULTS: In osteoporotic populations, the prevalence of 25(OH) vitamin D concentration <12 ng/ml ranged from 12.5% to 76%, while prevalence rates reached 50% to 70% of patients with a history of fracture(s) using a cut-off of 15 ng/ml. In post-menopausal women, the prevalence of 25(OH) vitamin D concentrations

Psychometric validation of an overactive bladder symptom and health-related quality of life questionnaire: the OAB-q.

OBJECTIVE: Overactive bladder (OAB) is characterized by symptoms of urinary frequency and urgency, with and without incontinence, and has been shown to have significant impact on health-related quality of life (HRQL). Currently, no OAB-specific questionnaires exist to evaluate all symptoms of OAB; thus we sought to evaluate the psychometric properties of a newly developed OAB HRQL questionnaire. METHODS: The 33-item, self-administered OAB-q contains a symptom bother and HRQL scale. Both the OAB-q and SF-36 were completed by participants from two sources: (1) a community sample who screened positive for OAB in a random-digit dial telephone survey and participated in a clinical validation study (n = 254); and (2) a clinical study of patients' seeking treatment for OAB symptoms (baseline assessment) (n = 736). Item and exploratory factor analysis were performed to assess the subscale structure of the questionnaire. Psychometric evaluation was conducted to assess reliability and validity. RESULTS: Seventy-four percent of the sample were women with mean age of 58.5. Participants with continent and incontinent symptoms reported significantly greater symptom bother and HRQL impact than normal participants. Significant differences were present among all patient groups in all OAB-q subscales (p < 0.0001) except sleep where the impact of continent and incontinent OAB was similar, but significantly worse than normal participants (p < 0.0001). Internal consistency was high with the subscale Cronbach alpha-values ranging from 0.86 to 0.94. CONCLUSION: The OAB-q is a reliable and valid instrument that discriminates between normal and clinically diagnosed continent and incontinent OAB participants. The OAB-q demonstrates that both continent and incontinent OAB symptoms cause significant symptom bother and have a negative impact on HRQL.

Clinical trial on the efficacy and safety of different diclofenac formulations: multiple-unit formulations compared to enteric coated tablets in patients with activated osteoarthritis.

This double-blind, randomised, multicentre study investigated the efficacy and safety of two different dosages of a diclofenac sodium dual release capsule (150 mg or 75 mg once daily) in comparison to a standard treatment with enteric coated tablets (50 mg t.i.d.) and placebo in patients with activated osteoarthritis. Pain relief as the main efficacy variable was measured through 24 hours by means of a Visual Analogue Scale at baseline and on five assessment days during the 12 weeks of treatment. Efficacy was observed in all treatment groups with a statistically significant difference between the verum groups and placebo. The overall safety and tolerability of the active treatments was good. For the 75 mg group, a lower incidence of liver and biliary system-related side effects was reported. Considering efficacy, safety, and compliance aspects, the once daily administration of diclofenac sodium 75 mg dual release capsule is the appropriate dosage regimen for mid- and long-term treatment of osteoarthritis.

Phytochemical characterization of various St. John's Wort extracts.

Hypericum perforatum is used for its beneficial effects on the nervous system, especially as an antidepressant. Owing to the fast-growing St. John's Wort market in the United States, more and more products (herb and extract) are sold at different quality levels. The amount of well-dried H. perforatum plant material is limited and other species with differing composition and no proof of efficacy are sold. The generally used method for standardization--spectrophotometric determination of naphthodianthrones--is not as selective as a high-performance liquid chromatographic (HPLC) method and can be manipulated (eg, by adding colorants). The different analytical methods used for characterization of St. John's Wort are discussed and a combined spectrophotometric and HPLC method is proposed. Qualitative and quantitative ranges for pharmaceutically relevant compounds of St. John's Wort extracts are cited and the composition of European, US, and Far East extracts is detailed. The influence of the production process, which involves extraction solvents, on phytochemical composition is also described.

Selective HPLC Analysis of n-Alkyl Hydroperoxides up to C(18)H(38)O(2).

A series of n-alkyl hydroperoxides are separated by HPLC and detected by their postcolumn reaction with horseradish peroxidase and p-hydroxyphenylacetic acid (HPAA) to yield a fluorescent product; several secondary and tertiary hydroperoxides, some 1-hydroxyalkyl hydroperoxides, and a few branched hydroperoxides are also examined. n-Alkyl hydroperoxides up to at least C-18 react with the enzyme with only minimally reduced efficiency at greater alkyl chain length. The effects of the column, the eluent, and the pH of the sample reaching the detector are described. The detection limit with gradient elution ranges from 0.4 µmol L(-)(1) for n-hexyl hydroperoxide to 1 µmol L(-)(1) for n-octadecyl hydroperoxide.

Structural analysis and localization of the carbohydrate moieties of a soluble human interferon gamma receptor produced in baculovirus-infected insect cells.

A soluble form of the human interferon gamma receptor that is required for the identification of interferon gamma antagonists was expressed in baculovirus-infected insect cells. The protein carried N-linked carbohydrate and showed a heterogeneity on denaturing polyacrylamide gels. We investigated the utilization of the potential sites for N-linked glycosylation and the structure of the carbohydrate moieties of this soluble receptor. Amino acid sequence analysis and ion spray mass spectrometry revealed that of the five potential sites for N-linked glycosylation, Asn17 and Asn69 were always utilized, whereas Asn62 and Asn162 were utilized in approximately one-third of the protein population. Asn223 was never found to be glycosylated. The soluble receptor was treated with N-glycosidase F and the oligosaccharides released were analyzed by matrix-assisted laser desorption mass spectrometry, which showed that the protein carried six types of short carbohydrate chains. The predominant species was a hexasaccharide of molecular mass 1,039, containing a fucose subunit linked to the proximal N-acetylglucosamine residue: [formula: see text]

Structural fluctuations between two conformational states of a transmembrane helical peptide are related to its channel-forming properties in planar lipid membranes.

Putative transmembrane helices of membrane proteins in general and channel proteins in particular often contain proline residues which may induce a bend into an otherwise regular helical structure. Here we show by fluorescence-energy-transfer measurements and molecular-dynamics calculations that, in the case of synthetic bilayer-spanning helical polypeptides, a proline-induced bend in a helix acts as a flexible element mediating rigid body motions of the helical segments. Most important, such structural fluctuations in the transmembrane helices seem to play a functional role in the formation of ionic channels in planar lipid bilayers and biological membranes.

The Structures of some Triterpene Saponines from Anagallis arvensis.

5 new triterpene saponines, named anagalloside A, B, C, and desglucoanagalloside A and B, were isolated from the herb of ANAGALLIS ARVENSIS L. (Primulaceae). The structures of the saponines were elucidated by (1)H-, (13)C-NMR spectroscopy, FAB-MS, and the methylating and combined methylating-ethylating analysis methods.

[Clinical and toxicologic study of axillary plexus block with prilocaine or mepivacaine]. / Klinische und toxikologische Untersuchung zur axillären Plexusblockade mit Prilocain oder Mepivacain.

A prospective controlled double-blind study was designed to compare the efficacy and the toxicity of mepivacaine and prilocaine for the axillary blockade of the brachial plexus. Twenty patients in each group received 40 ml of either 1.5% mepivacaine or 1.5% prilocaine. The sensory and the motor blockade achieved in both groups were comparable. The mean plasma levels attained following mepivacaine were significantly higher than those attained after prilocaine (peak-plasma level (mean +/- SD): 2.02 +/- 0.28 micrograms/ml vs 5.37 +/- 1.83 micrograms/ml). In four patients of the mepivacaine group, the plasma levels entered the toxic range. While no methemoglobinemia could be detected following mepivacaine, four of the patients receiving prilocaine developed methemoglobinemia with a maximum level of 10%. No complications due to methemoglobinemia were observed in any of the patients. Because of its significantly lower toxic potential, prilocaine seems to be the better local anaesthetic for axillary blockade of the brachial plexus.

Efficient HPLC separation of N-p-nitrophenylglycosylamines derived from complex oligosaccharide mixtures. Human orosomucoid as a model.

With human orosomucoid as model compound, a new method was developed to separate neutral oligosaccharides as N-p-nitro-phenylglycosylamines. Asialo orosomucoid was prepared by treatment with neuraminidase and purified by size exclusion HPLC on TSK 2000 SW. Oligosaccharides were isolated by reversed phase HPLC on Hamilton PRP-1 after hydrazinolysis and re-N-acetylation. Glycosylamination was performed with p-nitroaniline in DMSO-formic acid-water, where the whole mixture of oligosaccharide derivatives was isolated by reversed phase HPLC on Hamilton PRP-1 and separated into single glycosylamines on Shandon Hypersil ODS. The purified glycosylamines could be methylated by a new rapid method with sodium hydroxide and methyl iodide in DMSO, isolation and purification were carried out on Hamilton PRP-1 and Spherisorb ODS 2, respectively, as described for the glycosylamines. Preparative scale HPLC separations were performed on analytical columns using repetitive collection mode and automatic sample loading by means of a peristaltic pump operated by the HPLC controller. The purified glycosylamines can be used for sugar analyses or, after permethylation, for methylation analyses or related procedures.