Inorganic substances and their uses in Nikolaos Myrepsos' Dynameron. Recent applications in modern therapy.

Inorganic compounds have been known and used since antiquity. Dynameron is the largest Byzantine medical manuscript divided into 24 sections, in accordance with the letters of the Hellenic alphabet, which contains 2667 recipes. The majority of them contain ingredients of plant origin, followed by animal origin, while fewer inorganic substances are quoted. In the present study, the latter ones are listed. Moreover, the information on the uses of inorganic ingredients in the treatment of many diseases in the late Byzantine era is presented and their evaluation in light of the modern Pharmacology and Toxicology.

Remedies of animal origin and their indications in Nikolaos Myrepsos׳ Dynameron.

ETHNOPHARMACOLOGICAL RELEVANCE: Dynameron is a Byzantine medical compendium, divided into 24 sections, in accordance with the letters of the Greek alphabet. Being the largest medical and pharmaceutical book ever written in Byzantium, Dynameron contains 2667 recipes intended to treat many pathological conditions. A lot of information convey to us through prescriptions. In addition to plants, Nikolaos Myrepsos proposes the use of many animals, animal parts and animal by-products, for the treatment of various diseases. This article presents for the first time a full account of the animal products included in Dynameron. AIM OF THE STUDY: In continuation to our previous studies, this paper focuses on the use of animal products in composite medicines described in Dynameron. An effort was made to trace down the use of similar or identical animal products in texts of earlier medical writers. Recording recipes with animals or animal products intended for use in everyday medical practice highlights the timeless belief in their healing properties. MATERIALS AND METHODS: Our main source of material is the recent digital edition of Nikolaos Myrepsos' Dynameron. This huge treatise was written in the 13th century and reflects in many ways the long medical tradition of the Greek, the Hellenistic and the Roman eras, having also received influences from the materia medica of Arabic medicine. In addition, information from dictionaries and databases were cross-checked to confirm and classify the animals and their products and to identify them. For the various pathological conditions these products are meant for, we have used the current medical terminology. RESULTS: In the present study, we could identify the therapeutic use of 93 animals. In several instances, Myrepsos suggests the use of specific organs of an animal, and for that reason he includes in his treatise 16 anatomical parts of different animals. Moreover, Dynameron comprises also 34 animal by-products, such as milk and honey. Medicines of animal origin are used in recipes concerning diseases of the respiratory, the digestive, the cardiovascular and the urinary system, as well as gynecological diseases, and ailments of the eyes, the ears and the skin. CONCLUSIONS: Of the 2667 recipes of Dynameron, 344 recipes contain medicines of animal origin, which can be detected in totally 769 citations. In addition, 626 citations for animal by-products are found in 268 recipes. Honey and milk are quoted in 2136 recipes, mostly as excipients. Dietary instructions are present on many occasions, reflecting the attitude for a healthy everyday life, similar to the modern beliefs pertaining to food as an essential factor for a good health.

Saffron (Crocus sativus) intake provides nutritional preconditioning against myocardial ischemia-reperfusion injury in Wild Type and ApoE(-/-) mice: Involvement of Nrf2 activation.

BACKGROUND AND AIMS: Saffron is an antioxidant herbal derivative; however, its efficacy as a nutritional cardioprotective agent has not been fully elucidated. We investigated the cardioprotective properties of a standardized saffron aqueous extract (SFE) against ischemia/reperfusion (I/R) injury in Wild-Type (WT) and ApoE(-/-) mice and the underlying molecular mechanisms. METHODS AND RESULTS: WT and ApoE(-/-) mice were subjected to 30 min I and 2 h R, with the following per os interventions for 4 weeks: 1) WT Control Group, receiving Water for Injection (WFI); 2) WT Crocus Group, receiving SFE at a dose of 60 mg/kg/day; 3) WT Crocus + Wort group, receiving SFE as described above and wortmannin at a dose of 60 µg/kg bolus 15 min before R; 4) ApoE(-/-) Control Group, receiving WFI; 5) ApoE(-/-) Crocus Group, receiving SFE at a dose of 60 mg/kg/day and 6) ApoE(-/-) Crocus + Wort: receiving SFE as described above and wortmannin at a dose of 60 µg/kg bolus, 15 min before R. Ischemic area/area at risk (I/R%) ratio was measured. Blood samples and ischemic myocardial tissue were collected at the 10th min of reperfusion for assessment of troponin I, malondialdehyde (MDA), nitrotyrosine (NT), p-eNOS, eNOS, p-Akt, Akt, p-p42/p-p44, p-GSK3ß, GSK3ß, IL-6, Nrf2, HO-1 and MnSOD expression. The effect of SFE on Nrf2 expression was also evaluated in vitro. SFE reduced infarct size in WT (16.15 ± 3.7% vs 41.57 ± 2.48%, ***p < 0.001) and in ApoE(-/-) mice (16.14 ± 1.47% vs 45.57 ± 1.73%, ***p < 0.001). The administration of wortmannin resulted in partial inhibition of the infarct size limitation efficacy of SFE (in both WT and Apo-E(-/-) mice). Mice receiving SFE showed increased levels of eNOS, p-Akt, p-ERK1/2, p-44/p-42 and p-GSK3ß-Ser9 and reduced expression of IL-6 and iNOS; furthermore, SFE reduced the levels of MDA and NT. SFE induced Nrf2 expression and its downstream targets, HO-1 and MnSOD in the myocardium of the treated animals, and induced Nrf2 expression in vitro in a dose-dependent manner. CONCLUSIONS: SFE limits myocardial infarction in Wild-Type and ApoE(-/-) mice in a multifaceted manner including activation of Akt/eNOS/ERK1/2/GSK3-ß and through Nrf2 pathway, bestowing antioxidant protection against I/R.

Ethnopharmacological approach to the herbal medicines of the "Elements Alpha to Delta" in Nikolaos Myrepsos׳ Dynameron. Part II.

ETHNOPHARMACOLOGICAL RELEVANCE: Dynameron is a Byzantine medical compendium, divided into 24 sections, the "Elements", containing 2667 recipes, most of which inherited by previous physicians of the classic ancient Greek and Hellenistic, and imperial Roman periods. AIM OF THE STUDY: In continuation to our previous study concerning the first and largest chapter of the "Element Alpha" of Nikolaos Myrepsos׳ Dynameron (Valiakos et al., 2015), this paper focuses on the plants quoted in the recipes of the eight following chapters entitled "About Salts", "About Honeypacks" and "About Spreads", all belonging to the same "Element Alpha"; "About Antitussives" and "About Suppositories" belonging to the "Element Beta"; "About women's Cathartics" belonging to the "Element Gamma"; "About Drossaton" and "About Diachrisma", both belonging to the "Element Delta". MATERIALS AND METHODS: Our main primary source material was the codex kept in the National Library of France (in Paris) under the number grec. 2243, which is the older and larger codex of Dynameron (Valiakos et al., 2015). RESULTS: The present study led us to the interpretation of 277 plants under different names, among which we recognized 57 medicinal plants listed by the European Medicines Agency, one of them with negative monograph (i.e. Chelidonium majus). In addition, there are identified taxa related to those quoted by EMA as herbal medicines. The plants appearing in the examined Elements belong to various families of which the most frequent are: Apiaceae 10.11%; Lamiaceae 7.22%; Asteraceae 6.86%; Rosaceae 6.5% and Fabaceae 6.14%. CONCLUSIONS: A total of 277 species have been catalogued, most of which are referred in our previous publication (Valiakos et al., 2015). Among them, 56 plants still play a very important role in medical practice, as they are used as traditional herbal medicines (www.ema.eu). This evidence is a proof that the use of medicinal plants remains valuable from the ancient times until today. The recipes, in contrast to older medical compendia, contain precise measurements of ingredients and dosages for every drug, which seem to reflect empirical logic.

Ethnopharmacological approach to the herbal medicines of the "Antidotes" in Nikolaos Myrepsos׳ Dynameron.

ETHNOPHARMACOLOGICAL RELEVANCE: This paper focuses on the plants quoted in the recipes of the first chapter entitled "About the Antidotes" belonging to the first and largest section "Element Alpha" of Nikolaos Myrepsos׳ Dynameron, a medieval medical manuscript. Nikolaos Myrepsos was a Byzantine physician at the court of John III Doukas Vatatzes at Nicaea (13th century). He wrote in Greek a rich collection of 2667 recipes, the richest number known in late Byzantine era, conventionally known as Dynameron and divided into 24 sections, the "Elements". The only existing translation of this work is in Latin, released in 1549 in Basel by Leonhart Fuchs. Since no other translation has ever been made in any language, this work still remains poorly known. MATERIALS AND METHODS: Our primary source material was the codex written in 1339 and kept in the National Library of France (in Paris) under the number grec. 2243. For comparison, all the other codices, which contain the entire manuscript, have also been studied, namely the codices EBE 1478 (National Library of Greece, Athens), grec. 2237 and grec. 2238 (both in Paris), Lavra Ε 192 (Mont Athos, Monastery of Megisti Lavra), Barocci 171 (Oxford) and Revilla 83 (Escorial). RESULTS: The exhaustive study of the "About the Antidotes" led us to the interpretation of 293 plant names among which we recognized 39 medicinal plants listed by the European Medicines Agency, (Herbal Medicines, www.ema.eu); the therapeutic indications of some of them provided by Myrepsos were similar or related to their current ones, as given in their monographs. The plants belong to various families of which the most frequent are: Apiaceae 10.6%; Lamiaceae 9.2%; Asteraceae 8.9%; Fabaceae 6.8% and Rosaceae 5.1%. The most frequently mentioned plants even under several different names are the following: Apium graveolens L., Crocus sativus L., Nardostachys jatamansi (D. Don) DC., Zingiber officinale Roscoe, Rosa centifolia L., Syzygium aromaticum (L.) Merr. & L.M. Perry, Papaver somniferum L., Costus sp., Petroselinum crispum (Mill.) Fuss, Anethum graveolens L., Foeniculum vulgare Mill., Daucus carota L. CONCLUSIONS: This research led us to the conclusion that the content of "About the Antidotes" is a valuable source for the study of recipes based mainly on medicinal plants, most of them inherited from classic ancient Greek and Hellenistic periods.

Antimicrobial activity of essential oils of Xylopia aethiopica.

Xylopia aethiopica is a medicinal plant of great repute in West Africa which produces a variety of complex chemical compounds. The fresh and dried fruits, leaf, stem bark and root bark essential oils showed various degrees of activity against the gram positive bacteria, Bacillus subtilis and Staphylococcus aureus, the gram negative bacteria Pseudomonas aeruginosa and the yeast-like fungus Candida albicans, using the cup plate method. However, none of the oils showed activity against Escherichia coli.

Analysis of the essential oil of Origanum dubium growing wild in Cyprus. Investigation of its antioxidant capacity and antimicrobial activity.

The volatile composition of Origanum dubium in two different maturation stages has been studied. The essential oils were obtained by hydrodistillation in a modified Clevenger-type apparatus, and their analyses were performed by GC and GC-MS. Identification of the components was made by comparison of mass spectra and retention indices with literature records and by co-chromatography with authentic compounds. Carvacrol was shown to be the main constituent. The essential oils were evaluated for antimicrobial activity and proved to be active against all tested microorganisms. Furthermore, their potential antioxidant activity was investigated and found to be significant in scavenging O2-. The samples were further evaluated for inhibition of soybean lipoxygenase LOX and showed high inhibitory activity.

Biocatalytic preparation of acylated derivatives of flavonoid glycosides enhances their antioxidant and antimicrobial activity.

Enzymatic synthesis of acylated derivatives of a monosaccharidic flavonoid chrysoeriol-7-O-beta-D-(3''-E-p-coumaroyl)-glucopyranoside as well as of a disaccharidic flavonoid chrysoeriol-7-[6'''-O-acetyl-beta-D-allosyl-(1-->2)-beta-D-glucopyranoside], isolated from Greek endemic plants, was performed using an immobilized Candida antarctica lipase in non-toxic organic solvents. The influence of the reaction parameters such as the molar ratio of acyl donor to flavonoid, as well as the nature of the acyl donor, on the performance of the biocatalytic process was pointed out using the acylation of naringin as a model reaction. With vinyl laurate as acyl donor, the highest conversion was observed at relatively high molar ratio (>or=10), using acetone as solvent. Lipase exhibits specificity towards primary alcohol of the glucose moiety of both flavonoid glycosides. The introduction of an acyl group into glucosylated flavonoids significantly improved their antioxidant activity towards both LDL and serum model in vitro. Furthermore, the acylated derivative of disaccharidic flavonoid increased its antimicrobial activity against two Gram-positive bacteria.

Effect of nitrogen concentration of the nutrient solution on the volatile constituents of leaves of Salvia fruticosa Mill. in solution culture.

Essential oils from hydroponically cultivated Salvia fruticosa were analyzed by GC-MS techniques. Three different levels of nitrogen (100, 150, and 200 mg/L) were used in the nutrient solution for the cultivation, using the nutrient film technique. A total of 79 compounds were identified, and qualitative and quantitative differences have been observed between the samples collected at full bloom (flowering stage) and at the end of the seed formation stage. 1,8-Cineole, beta-caryophyllene, and viridiflorol were the predominant constituents in most cases. 13-epi-Manool was identified by using GC parameters and mass spectrum fragmentation pattern, whereas labd-7,13-dien-15-ol, a labdane type diterpene, was identified for the first time in the genus Salvia, using GC parameters and an authentic sample. The results obtained from GC-MS analyses were submitted to chemometric analysis.

Antimicrobial activity of acidic xylo-oligosaccharides produced by family 10 and 11 endoxylanases.

Acidic oligosaccharides were obtained from birchwood xylan by treatment with a Thermoascus aurantiacus family 10 and a Sporotrichum thermophile family 11 endoxylanases. The main difference between the products liberated by xylanases of family 10 and 11 concerned the length of the products containing 4-O-methyl-D-glucuronic acid. The xylanase from T. aurantiacus liberate from glucuronoxylan an aldotetrauronic acid as the shortest acidic fragment in contrast with the enzyme from S. thermophile, which liberated an aldopentauronic acid. Acidic xylooligosaccharides were separated from the hydrolysate by anion-exchange and size-exclusion chromatography (SEC) and the primary structure was determined by 13C NMR spectroscopy. The acidic xylo-oligosaccharides were tested against three Gram-positive and three Gram-negative aerobically grown bacteria, as well as against Helicobacter pylori. Aldopentauronic acid was proved more active against the Gram-positive bacteria and against H. pylori.

A chemotaxonomic investigation of volatile constituents in Stachys subsect. Swainsonianeae (Labiatae).

The composition of the previously unknown essential oils of Stachys subsect. Swainsonianeae (Labiatae) are investigated by GC-MS analyses. The group includes the taxa Stachys ionica, St. spruneri, St. swainsonii ssp. swainsonii, St. swainsonii ssp. argolica, St. swainsonii ssp. melangavica and St. swainsonii ssp. scyronica, all endemic to Greece. Eighteen populations were analyzed, and more than two hundred components were identified. Statistical analyses of oil components separated St. ionica, due to its high amount of (E)-nerolidol, high amount of alpha-cadinol and low amount of (+)-(E)-caryophyllene. The rest of the taxa forms a coherent group where (+)-(E)-caryophyllene, delta-cadinene and partly beta-elemene, (+)-caryophyllene oxide and (+)-limonene predominate but each of them never exceeds 20% of the total. Essential oil composition reflects current taxonomy and phytogeography of the group with a somewhat isolated position for St. ionica. Similarities in morphology and essential oil composition of the St. swainsonii group and St. spruneri may be the result of extensive gene flow in the past. Evidence from volatile oil compounds that may attribute to St. swainsonii ssp. melangavica a hybrid origin from crosses between St. spruneri and St. swainsonii ssp. swainsonii/scyronica group is scarce.

Composition and antimicrobial activity of the essential oil of Scutellaria albida ssp. albida from Greece.

Steam distilled essential oil from aerial parts of Scutellaria albida ssp. albida was analyzed by GC and GC/MS. Fifteen compounds were identified of which linalool (52.63%) and trans-nerolidol (9.03%) were the major constituents. Furthermore, the oil was tested against four bacteria and two yeasts and was found to be moderately active against all microorganisms tested.

Inhibition of prostaglandin E2 and leukotriene C4 in mouse peritoneal macrophages and thromboxane B2 production in human platelets by flavonoids from Stachys chrysantha and Stachys candida.

Seven flavonoids of Stachys chrysantha and Stachys candida have been isolated. The structures of the compounds were elucidated by spectroscopic methods, particularly highfield NMR spectroscopy. The effects of the methanol extracts of these two endemic Greek Stachys sp. and their main flavonoids were examined on arachidonic acid (AA) metabolism in the cellular system (mouse peritoneal macrophages and human platelets). Their cytotoxicity on cells was also investigated. Most samples assayed did not exhibit any significant effect on prostaglandin E2 (PGE2)-release from calcium ionophore-stimulated mouse peritoneal macrophages. Only chrysoeriol-7-O-beta-(3''-E-p-coumaroyl)-glucopyranoside, at the highest non-cytotoxic dose (50 microM), inhibited the release of PGE2, but this effect is not statistically significant. The release of leukotriene C4 (LTC4) by mouse peritoneal macrophages stimulated with calcium ionophore was inhibited by a crude extract of S. chrysantha, with an IC50 value of 34.3 microg/ml. Xanthomicrol (IC50 = 29.2 microM) and chrysoeriol-7-O-beta-D-(3''-E-p-coumaroyl)-glucopyranoside (IC50 = 11.1 microM) also inhibited the release of LTC4, although it showed less potency than the reference compound nordihydroguaiaretic acid (NDGA) (IC50 = 2 microM). However, most samples assayed showed a significant effect on thromboxane B2 (TXB2)-release from calcium ionophore-stimulated human platelets, with inhibition percentages slightly lower than the reference drug ibuprofen (IC50 = 7 microM). The IC50 values are: crude extract of S. candida 23.3 microg/ml; crude extract of S. chrysantha 23.1 microg/ml; xanthomicrol 28.8 microM; calcycopterin 2.66 microM and chrysoeriol-7-O-beta-D-(3''-E-p-coumaroyl)-glucopyranoside 8.8 microM. Our results indicate that the selective inhibition of TX-synthase enzyme may be the primary target of action of most of these samples, and one of the mechanisms through which thus exert their antiinflammatory effects.

Sesquiterpene lactones from Centaurea thessala and Centaurea attica. Antifungal activity.

The aerial parts of Centaurea thessala ssp. drakiensis and C. attica ssp. attica afforded, in addition to several known sesquiterpene lactones, two new eudesmanolides, 4-epi-sonchucarpolide and its 8-(3-hydroxy-4-acetoxy-2-methylene-butanoyloxy) derivative and one new eudesmane derivative, named atticin. The in vitro antifungal activity of most compounds was tested against nine fungal species, using the micro-dilution method. All the compounds tested showed great antifungal activity.

Composition and antibacterial activity of the essential oils of Stachys candida and S. chrysantha from southern Greece.

The chemical composition of the essential oils obtained from the aerial parts of Stachys candida and S. chrysantha were analysed with GC and GC/MS. Forty-two constituents were identified from the two oils and alpha-cadinol, manoyl oxide, caryophyllene oxide, epi-alpha-muurolol and (E)-caryophyllene were found to be the major components. Furthermore, the oils were tested against six Gram (+/-) bacteria and it was found that one of them exhibited significant antibacterial activity.

Antiphage properties of some Greek algae extracts.

The ethanol extracts of 35 algae collected in several Greek shores, were screened for antiphage properties against 6 bacteriophages: coliphages T 1, T 2, T 4, T 7, ΦX 174 and MS 2. Five samples were found to induce a mild antiphage activity.

[The Aconit described by Nicander and today]. / L'aconit chez Nicandre et de nos jour.

The herbal drug Aconite described by Nicander of Colophon (IInd century BC) is studied as to its symptoms and uses. The botanical identification presents some difficulties. There is a close correlation between the symptoms from Nicander's work and those cited by homeopaths authors.