Biotransformation of Monoterpenoids by Suspension Cultures of Lavandula angustifolia

Abstract Callus and suspension cultures of Lavandula angustifolia Mill. (Lamiaceae) were established and the effect of different culture media on growth rate was investigated. Terpenoids added to suspension culture to investigate their biotransformation. All samples were analyzed by gas chromatography (GC) and GC-mass spectroscopy (MS). Octan-1-ol, citronellol, linalool, borneol and geraniol were biotransformed products of octanal, citronellal, linalyl acetate, bornyl acetate and geranyl acetate, respectively. Citronellol, linalool, borneol, and menthol didn't change by L. angustifolia suspension cultures. Blue pigment production by cultures of L. angustifolia was also studied. Ester hydrolysis and oxidation were the main reactions which occurred in biotransformation process, which may be attributed to the presence of related or bifunctional enzymes. This technique is a possible way of the production of expensive or rare compounds from cheap and plentiful substrates

Root Cultures of Linum Species Section Syllinum as Rich Sources of 6-Methoxypodophyllotoxin

professor of Pharmacognosy, Tehran Faculty of Pharmacy, Tehran on the occasion of his birthday Linum spp. from section Syllinum are promising for the production of aryltetralin lignans like podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (MPTOX). MPTOX is a PTOX congener that has cytotoxic activity comparable with PTOX. In this study root cultures of Linum Bungei from section Dasyllinum, L. strictum from section Linastrum, L. album, L. mucronatum ssp. mucronatum and L. nodiflorum from section Syllinum were established and their MPTOX levels were investigated in 1000 ml flasks. Root cultures of L. mucronatum ssp. mucronatum and L. nodiflorum were used to examine cell growth and production of MPTOX during a culture period of 36 days in 250 ml flasks. Considerable amounts of MPTOX in root cultures (1000 ml flasks) of L. album (6 mg/100 g DW), L. mucronatum ssp. mucronatum (770 mg/100 g DW) and L. nodiflorum (91 mg/100 g DW) were detected while it wasn't detected in root cultures of L. Bungei and L. strictum. In time course experiments, the maximum amount of MPTOX in L. nodiflorum root culture was at day 16 with 480 mg/ 100 g DW and the maximum amount of MPTOX in L. mucronatum ssp. mucronatum root culture was at day 12 with 130 mg/100 g DW. The results showed that root cultures of Linum species from section Syllinum are rich sources of MPTOX and since this lignan has remarkable cytotoxic activity, it can be used as a precursor for the production of antitumor agents

ATR-IR fingerprinting as a powerful method for identification of traditional medicine samples: a report of 20 herbal patterns

Abstract Background and objectives: Attenuated total reflectance-inferared (ATR-IR) spectra can be used as a non-invasive fingerprinting approach in quality control of herbal samples. Methods: Twenty versatile herbal samples were subjected to attenuated total reflectance-inferared (ATR-IR) spectroscopy followed by different clustering methods in order to determine by which method more reasonable classifications would be obtained. Results: All classification methods (K-means, HCA, PCA and SOM) were able to discriminate the two medicinal seeds, Hyocyamus niger and Peganum harmala from other herbal samples. Similarly, the starch samples were clustered in a reasonable method. In HCA, one cluster included three types of starch samples: Zea mays, Oryza sativa and Triticum aestivum. All the four classification methods were able to separate Solanum tuberosum starch from other starch samples. HCA and SOM, were able to classify leaf samples Origanum vulgare and Melissa officinalis belonging to Lamiaceae family, in one category. Crocus sativus and its adulterant Carthamus tinctorius flowers were identified by PCA, HCA and SOM as different categories. Conclusion: The result of this study can be utilized for identification and quality control of traditionally used medicinal plant samples in an unknown sample powder. Such data could be the basis for preparing a data bank on Iranian medicinal samples which in turn is used as a simple, fast and reliable method for characterization of herbal powders in Pharmacopoeias

ATR-IR fingerprinting as a powerful method for identification of traditional medicine samples: a report of 20 herbal patterns

Background and objectives: Attenuated total reflectance-inferared (ATR-IR) spectra can be used as a non-invasive fingerprinting approach in quality control of herbal samples. Methods: Twenty versatile herbal samples were subjected to attenuated total reflectance-inferared (ATR-IR) spectroscopy followed by different clustering methods in order to determine by which method more reasonable classifications would be obtained. Results: All classification methods (K-means, HCA, PCA and SOM) were able to discriminate the two medicinal seeds, Hyocyamus niger and Peganum harmala from other herbal samples. Similarly, the starch samples were clustered in a reasonable method. In HCA, one cluster included three types of starch samples: Zea mays, Oryza sativa and Triticum aestivum. All the four classification methods were able to separate Solanum tuberosum starch from other starch samples. HCA and SOM, were able to classify leaf samples Origanum vulgare and Melissa officinalis belonging to Lamiaceae family, in one category. Crocus sativus and its adulterant Carthamus tinctorius flowers were identified by PCA, HCA and SOM as different categories. Conclusion: The result of this study can be utilized for identification and quality control of traditionally used medicinal plant samples in an unknown sample powder. Such data could be the basis for preparing a data bank on Iranian medicinal samples which in turn is used as a simple, fast and reliable method for characterization of herbal powders in Pharmacopoeias

The Use of Frozen Autogenous Bone Flap for Cranioplasty

Background: The artificial methods of cranioplasty such as using metals or alloplastic materials have some disadvantages, comparning with autogenic bone flaps. We tried to show that the autogenic flaps have less complications when used in cranioplasty.
 Methods: With good sealing of bone flap after extraction and preserving in -70 to -800 C, in Immounology Department, the autogenic bone was fixed in the previous site.
 Results: From 10 patients, one of them developed infection and osteomyelitic bone was extracted. No bone resorption was detected.
 Background: Comparing with other studies of autogenous bone flap cranioplasty, we have similar rate of complication. In other studies, the rate of infection was almost equal to our results. So using autogenous bone in our center is advisable.
 Key words: cranioplasty, autogenous bone, frozen bone, infectio

Quantification of Aryltetralin Lignans in Linum album Organs and In Vitro Cultures: Lignans from Linum album

Aprocedure was developed for rapid in vitro germination of Linum album seeds by scarification and exposing to gibberellic acid (GA3) and kinetin (Kn). Concentrations of podophyllotoxin (PTOX) and related aryltetralin lignans α-peltatin, β-peltatin, 5’-demethoxy-6-methoxypodophyllotoxin and 6-methoxypodophyllotoxin (MPTOX) in L. album fresh plant organs were determined by HPLC. A degree of variation was observed in lignan content in different plant parts and growth stages. It was found that PTOX is predominant in productive organs and leaves. The highest amount of PTOX (0.651 mg/100g dry wt.) and MPTOX (0.670 mg/100g dry wt.) and the total quantified lignans were found in the capsules. In vitro cultures were studied for lignans followed by high productive cell line selection. Calli cultures were more productive for MPTOX than PTOX, while suspension cells accumulate comparable amounts of PTOX and MPTOX. The highest amount of PTOX (0.301% mg/g dry wt.) was found in suspension originated immobilized cultures

Analysis of volatiles and 18S rRNA gene of Haplophyllum canaliculatum in in vitro cultures

Background and objectives:  Haplophyllum canaliculatum isan endemicand endangeredIranian plant from Rutaceae family. The object of this work was to study the volatile production in established shoot and callus cultures of Haplophyllum canaliculatum as well as isolation, identification and sequencing of 18S rRNA gene from callus culture. Methods: Shoot and callus cultures of H. canaliculatum were established from seedlings and shoot cultures, respectively. Both cultures were transferred to MS medium supplemented with α-naphthalene acetic acid (α-NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (Kn). Volatiles from fresh callus and shoot cultures were extracted and analyzed by GC/MS. For 18S rRNA gene study, DNA content was extracted using PCR procedure. The study of sequence similarities was performed using NCBI database and GeneDoc software. Results: GC/MS analysis of H. canaliculatum showed that shoot cultures mainly contained piperitone (10.92%), and β-caryophyllene (12.67%) in addition to three alkaloids, while calli cultures of H. canaliculatum mainly contained methylated salicylate (31.55%), alkane structures like tetradecane (24.31%) and hexadecane (12.95%). Gene analysis showed 98% homology with certain species of Rutaceae, Meliaceae, Simaroubaceae, Burseraceae and Cneoraceae. Conclusions: Our results showed that the hydrocarbon in addition to methyl salicylate biosynthetic pathway in calli cultures and terpene as well as alkaloid biosynthetic pathway were active in H. canaliculatum shoot cultures. Moreover, the obtained sequences could be used as a “DNA barcoding” tool through the concept of one sequence one species for the practical identification of this species

Evaluation of thermal-stress on the accumulation of podophyllotoxin in shoot in vitro cultures of Linum persicum

Linum persicum is an endangered plant native to Iran, from Linaceae family. Phytochemical analysis has shown that L. persicum in vitro cultures contain podophyllotoxin (PTOX), the pharmacological precursor for anticancer drugs such as etoposide, etopophos, and teniposide. Shoot culture of L. persicum was established from callus cultures in MS medium containing GA3. A study was designed to investigate the effects of thermal-stress treatments (TST) on the shoot growth pattern of L. persicum along with the accumulation of PTOX. Shoots were heat-stressed at 37, 40, 45, 50, and 55 °C for 3 h, and then transferred back to the growth room. In each thermal group after 2, 3, and 4 weeks, 4 flasks were extracted for PTOX analysis. HPLC analysis (