Simultaneous identification and quantification of pentacyclic triterpenoids and phenolic compounds from the leaves of Boswellia serrata using LC-MS/MS tandem mass spectrometry.

Boswellia serrata (B. serrata) is an important medicinal plant widely used as dietary supplements to provide a support for osteoarthritic and inflammatory diseases. The occurrence of triterpenes in leaves of B. serrata is very little or none. Therefore, the qualitative and quantitative determination of phytoconstituents (triterpenes and phenolics) present in the leaves of B. serrata is very much needed. The aim of this study was to develop an easy, rapid, efficient and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method for the identification and quantification of the compounds present in the leaves extract of B. serrata. The purification of ethyl acetate extracts of B. serrata was performed by solid phase extraction method, followed by HPLC-ESI-MS/MS analysis. Chromatographic parameters of the analytical method included negative electrospray ionization (ESI-) with a flow of 0.5 mL/min in gradient mode consisting of acetonitrile (A) and water (B) containing 0.1% formic acid, at 20 °C. Total 19 compounds (13 triterpenes and 6 phenolic compounds) were separated, and simultaneously quantified using a validated LC-MS/MS method with high accuracy and sensitivity. Good linearity was obtained with r2 > 0.973 in the calibration range. The overall recoveries were in a range between 95.78 and 100.2% with relative standard deviations (RSD) below 5% for the entire procedure of matrix spiking experiments. Overall, there was no ion suppression from the matrix. The quantification data showed that the total amount of triterpenes and phenolic compounds in the leaves of B. serrata ethyl acetate extract samples ranged from 14.54 to 102.14 mg/g and 2.14 to 93.12 mg/g of dry extract, respectively. This work provides, for the first time, a chromatographic fingerprinting analysis on the leaves of B. serrata. A rapid, efficient, and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and used for the both identification and quantification of triterpenes and phenolic compounds in the leaves extracts of B. serrata. The method established in this work can be used as quality-control method for other market formulations or dietary supplements containing leaf extract of B. serrata.

Molecular dynamics simulation and in vitro evaluation of herb-drug interactions involving dietary polyphenols and CDK inhibitors in breast cancer chemotherapy.

Dietary polyphenols such as quercetin and curcumin have been extensively administered to patients with cancer in the form of herbal supplements. They may have a synergistic anticancer effect; however, a risk of pharmacokinetic interactions with selective CDK-4/6 inhibitors that are metabolized by the CYP3A4 enzyme exists. Considering these pharmacokinetic aspects, the current study examined the effects of curcumin and quercetin on human CYP3A4 to ascertain CYP3A4-mediated herb-drug interactions with CDK inhibitors. In this study, using in silico methods and CYP3A4 inhibition kinetics in human liver microsomes and recombinant CYP3A4 enzymes, the effects of concentration-dependent inhibition of CYP3A4 by quercetin and curcumin on CDK inhibitors metabolism were examined. Based on our in-silico docking findings, curcumin and quercetin were considerably bound to CYP3A4 protein and displace CDK inhibitors from the CYP3A4 substrate binding domain. The IC50 values of curcumin and quercetin were 16.10 and 0.05 µM, respectively, for CYP3A4-mediated 1'-hydroxylation of midazolam. The dietary polyphenols prolonged the in vitro half-life of palbociclib and ribociclib by 6.4-fold and decreased their intrinsic microsomal clearance by approximately 4.6 times. Our findings indicate that curcumin and quercetin effectively cause herb-drug interactions and should be cautiously used to avoid therapeutic failure.

Quantitative estimation of total tannin, alkaloid, phenolic, and flavonoid content of the root, leaf, and whole plant of Byttneria herbacea Roxb.

Background: Byttneria herbacea Roxb., family Sterculiaceae, commonly called as 'Samarakhai' in local Odia language, is one of the reputed folklore medicinal herb. Its roots, leaves, and whole plant parts are reported for traditional use in the management of dysentery, sores, cuts, wounds, cholera, leucorrhoea, fractures, and sprains. Aim: The aim of the present work was to assess the total tannin content, total alkaloid content (TAC), total phenolic content (TPC), and total flavonoid content (TFC) in root, leaf, and whole plant of B. herbacea. Materials and methods: The amount of total tannins was analyzed using titrimetric method and total alkaloids by gravimetrical method. TPC was measured using Folin-Ciocalteu's method and calculated as gallic acid equivalents and the amount of total flavonoids by aluminum chloride colorimetric method and calculated as chrysin equivalents. Results: Tannin content was found maximum in the leaf (8.148% w/w) followed by whole plant (3.886% w/w) and root (1.553% w/w); similarly, TAC in the leaf (2.306% w/w) was more than those in root (0.814% w/w) and whole plant (1.319% w/w). The TPC of the methanolic extract of root (372.33 ± 14.29 mg/g) was more than whole plant (267.33 ± 7.63 mg/g); The TFC of the methanolic extract of leaf (620 ± 50 mg/g) was found maximum followed by root (553.33 ± 28.86 mg/g) and whole plant (536.66 ± 28.86 mg/g). Conclusion: The result of study emphasized presence of tannin, alkaloid, phenol, and flavonoid contents in the root, leaf, and whole plant of B. herbacea where the leaf was found to be richest source.

Investigation of Molecular Properties that Influence the Permeability and Oral Bioavailability of Major ß-Boswellic Acids.

BACKGROUND AND OBJECTIVES: Boswellic acids (BAs) include ß-boswellic acid (BA), 3-acetyl-ß-boswellic acid, 11-keto-ß-boswellic acid, 3-acetyl-11-keto-ß-boswellic acid, ß-boswellic alcohol, and 3-acetyl-11-hydroxy-ß-BA from Boswellia species, and are the main active ingredients of Boswellia serrata extracts (BSE). BSE have been used for the treatment of different inflammatory diseases; however, their pharmaceutical development has been severely limited by their poor oral bioavailability. The aims of this study were to investigate the molecular properties of six BAs, and to determine their experimental aqueous solubility, partition coefficient (Log P), gastrointestinal stability, adsorption-desorption kinetics, and permeability studies. METHODS: The physicochemical properties of six BAs were obtained from SMILES representations using ChemDraw, and MarvinSketch. The molecular properties were also determined experimentally. The permeability studies were performed using parallel artificial membrane permeability assay (PAMPA), and Caco-2 cells. RESULTS: The experimental Log P values of BAs correlated well (R2 = 0.94) with the calculated Log P values. Metabolic stability data confirmed that BAs were found to be unstable in simulated gastrointestinal fluids and intestinal S9 fractions. The apparent permeability (Papp) range of BAs in both PAMPA and Caco-2 for the apical (AP) to basolateral (BL) was in the range of 0.52 ± 0.05 × 10-6 to 2.84 ± 0.14 × 10-6cm/s. The efflux ratio of Papp (BL → AP) to Papp (AP → BL) for all BAs was < 2 in Caco-2 cells, suggesting greater permeability in the absorptive direction. Caco-2 cell adsorption studies confirmed the accumulation of BAs (35-55%) inside the enterocytes. These compounds exhibited a strong correlation between PAMPA and Caco-2 cell monolayer permeation data. CONCLUSIONS: The results of the present study have shown an empirical relationship between the molecular properties and intestinal absorption of BAs for the first time.

Evaluation of nutritional value and antioxidant activity of root and leaf of Samarakhai (Byttneria herbacea Roxb.): An extra pharmacopoeial herb.

BACKGROUND: Samarakhai (Byttneria herbacea Roxb.), family Sterculiaceae, is one of the reputed folklore medicinal herbs, found in many parts of India. Although consumed as a vegetable since long time, its root and leaves are not yet reported for its nutritive value and antioxidant activities. AIM: The aim of this study was to evaluate nutritional value and antioxidant potential of root and leaf of B. herbacea Roxb. MATERIALS AND METHODS: Nutritional parameters such as carbohydrate, fat, protein, energy value, calcium, iron, zinc, manganese, phosphorus and vitamin C were evaluated. Antioxidant activity was evaluated through three test methods, i.e., 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power (FRAP) and phosphomolybdenum assay. RESULTS: B. herbacea roots and leaves showed the presence of total carbohydrate 46.39 g/100 g and 40.12 g/100 g, total fat 0.63 g/100 g and 1.20 g/100 g, true protein 11.46 g/100 g and 10.49 g/100 g, energy content 237.07 kcal/100 g and 213.24 kcal/100 g, iron 821.10 ppm and 889.64 ppm, zinc 9.2 ppm and 47.98 ppm, manganese 329.86 ppm and 474.59 ppm, phosphorus 0.40 ppm and 0.10 ppm and calcium 4856.84 ppm and 14964.49 ppm, respectively. The half-maximal inhibitory concentration values of the methanol extract of root, leaf and ascorbic acid were found to be 217.25 µg/ml, 131.42 µg/ml and 178.88 µg/ml, respectively. In FRAP assay, antioxidant activity of methanol extract of leaf (129.15 µM) was found to be more than root (73.13 µM). CONCLUSION: B. herbacea root contains high amount of true protein, carbohydrate and energy value, while micronutrients such as iron, zinc, manganese and calcium are more in its leaf. Both roots and leaves exhibited potent antioxidant activity where the leaves possess more values than the roots.

Microperimetry biofeedback training in a patient with bilateral myopic macular degeneration with central scotoma.

Microperimetry-1 (MP-1) evaluation and MP-1 biofeedback training were done in a case of bilateral myopic macular degeneration with a central scotoma. Fixation behavior, location and stability of preferred retinal locus, eye movement speed, and mean sensitivity were assessed. The mean retinal sensitivities before, after and at 1-year after training in the right eye were 2.9 dB, 2.9 dB and 3.7 dB and in the left eye were 3.5 dB, 3.7 dB and 1.8 dB. The fixation point in the 2° gravitation circle, improved from 40% to 50% in the right eye and from 43% to 67% in the left eye. The average eye speed before, after and at 1-year after training in right eye were 0.19°/s, 0.26°/s and 0.25°/s and in left eye were 0.36°/s, 0.25°/s and 0.27°/s. Thus, biofeedback training using MP-1 can improve the visual function in patients with macular diseases and central scotoma.

Local anesthetic agents for vitreoretinal surgery: no advantage to mixing solutions.

PURPOSE: To compare the efficacy of lidocaine, bupivacaine, and a mixture of both in patients undergoing peribulbar anesthesia for vitreoretinal surgery. DESIGN: Cross-sectional study. PARTICIPANTS: Ninety patients. METHODS: Patients who underwent vitreoretinal surgery were randomized into 3 groups based on the peribulbar injection they received: lidocaine, bupivacaine, or a combination of lidocaine and bupivacaine. MAIN OUTCOME MEASURES: Time of onset of analgesia, akinesia, and intraoperative pain, if any, was noted. The efficacy of the block was graded from 0 to 5 depending on the adequacy of anesthesia and akinesia and the need for local supplementation. RESULTS: Mean times of onset (± standard deviation) of sensory blockade for the lidocaine, bupivacaine, and combination groups were 2.14±0.18, 2.19±0.13, and 2.17±0.11 minutes, respectively (P = 0.103). Mean times of onset (± standard deviation) of motor blockade for the lidocaine, bupivacaine, and combination groups were 3.04±1.81, 4.04±2.68, and 3.38±2.48 minutes, respectively (P = 0.255). Mean time of onset of intraoperative pain for the bupivacaine group, 149.33±46.33 minutes, was prolonged significantly compared with that of the combination group, 115.83±34.49 minutes, and that of the lidocaine group, 94.17±49.86 minutes (P < 0.001). Adequate anesthesia and akinesia (grade 5) were achieved in 56.7% of the patients in the bupivacaine group compared with 23.3% in the lidocaine group and 30% in the combination group (P = 0.049). CONCLUSIONS: In peribulbar anesthesia, 0.5% bupivacaine solution provides better quality of anesthesia than does combination 2% lidocaine and 0.5% bupivacaine in patients undergoing vitreoretinal surgery.

Transpupillary thermotherapy for circumscribed choroidal hemangioma: clinical profile and treatment outcome.

BACKGROUND AND OBJECTIVE: To report the outcome of treating circumscribed choroidal hemangioma (CCH) using transpupillary thermotherapy (TTT). PATIENTS AND METHODS: An infrared diode laser was used at 810 nm with a median spot size of 2.5 mm, power of 550 mW, and duration of 6.5 minutes; the median number of sittings was two. TTT spared the center of the macula when CCH involved the fovea. RESULTS: The mean age was 36 years (range: 13 to 63 years). CCH was subfoveal in 37.5% of the eyes and exudative retinal detachment was evident in all eyes. The tumor dimensions included thickness (median: 4.05 mm; range: 2.2 to 9.1 mm), horizontal diameter (median: 11 mm; range: 6 to 17.7 mm), and vertical diameter (median: 9.4 mm; range: 5.9 to 15.8 mm). After TTT, visual acuity improved in 44%, was maintained in 37%, and worsened in 19% of eyes. Likewise, tumor regression was achieved in thickness, vertical, and horizontal diameter in proportion of 20%, 15%, and 11%, respectively. The exudative retinal detachment resolved in 94% of eyes at the last follow-up. The median follow-up was 9.5 months. CONCLUSION: Macula-sparing TTT was effective in causing regression of CCH and resorption of associated exudative retinal detachment, and helped in preserving or improving vision in 81% of treated eyes.

Vitamin E supplementation may ameliorate oxidative stress in type 1 diabetes mellitus patients.

BACKGROUND: Increasing evidence in both experimental and clinical studies suggests that free radical mediated oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus. Proteins and lipids are among the prime targets for oxidative stress. In this study we evaluated oxidative stress in Type 1 Diabetes Mellitus (insulin dependent diabetes mellitus, IDDM) patients by estimating lipid peroxidation and the effect of vitamin E on oxidative stress and metabolic parameters. METHODS: A total of 40 children (20 Type 1 Diabetes Mellitus patients + 20 healthy controls) were examined in the study. Oxidative stress parameters malondialdehyde (MDA), antioxidants, reduced glutathione (GSH), vitamin E and metabolic parameters were studied. All the type 1 Diabetes Mellitus patients were supplemented with 600 mg/daily vitamin E for three months. After three months of supplementation all the parameters mentioned above were studied again. RESULTS: Reduced glutathione and vitamin E levels were lower and malondialdehyde levels were higher in Type 1 Diabetes Mellitus patients compared to healthy controls (p < 0.05). After supplementation with vitamin E in diabetic patients a significant decrease (p < 0.05) in MDA levels and significant increase in GSH (p < 0.05) and vitamin E (p < 0.05) levels were found. A negative correlation between MDA and vitamin E, between MDA and GSH and a positive correlation between vitamin E and GSH was found. Significant changes were not observed in metabolic parameters in Type 1 Diabetes Mellitus patients after vitamin E supplementation (p > 0.05). CONCLUSIONS: Vitamin E ameliorates oxidative stress in Type 1 Diabetes Mellitus patients and improves antioxidant defense system. However, vitamin E does not have any advantage for metabolic parameters.

Hydroxycut(®) (herbal weight loss supplement) induced hepatotoxicity: a case report and review of literature.

Use of supplement and alternative drugs continues to thrive and is becoming an increasing cause of concern since many of these substances may have unexpected or unexplained medical consequences. We present below the first reported case of hepatotoxicity from Hydroxycut(®) in Hawaii.

Transpupillary Thermotherapy for Juxtafoveal Leak in Central Serous Chorioretinopathy.

The authors describe the use of transpupillary thermotherapy (TTT) for juxtafoveal central serous chorioretinopathy (CSC) leak. A 45-year-old man with juxtafoveal leak developed fibrinous exudate in the macular area, demonstrated on optical coherence tomography. After one session of TTT, fibrinous exudate totally disappeared with improvement in vision to 20/40 from counting finger. Subretinal fluid got absorbed with obliteration of leak on fluorescein angiogram. TTT may be useful as a treatment option in juxtafoveal leak of CSC.

Blood salvage use in gynecologic oncology.

BACKGROUND: Blood salvage allows for collection and processing of surgical blood loss with the eventual reinfusion of washed red blood cells (RBCs) back to the patient. The use of blood salvage in patients undergoing surgery for malignancy is off-label. Controversy exists as to the risk of potential cancer dissemination resulting from the reinfusion of the processed blood, but no data are available to confirm this risk. Recent studies have demonstrated that filtering the salvaged blood using a leukoreduction filter (LRF) significantly decreases the number of cancer cells in the recovered RBC aliquot in a variety of cancer types. STUDY DESIGN AND METHODS: Patients on the gynecologic oncology service as part of the bloodless surgery program at Englewood Hospital and Medical Center from April 1998 to April 2007 were identified. Three patients that had reinfusion of cell salvage blood (all reinfusions were performed after filtration with a LRF) were studied further with emphasis placed on long-term outcomes. RESULTS: Two of the three patients did not show any evidence of metastases after surgery. The only patient that developed evidence of hematogenous progression had known liver metastases at the time of her initial diagnosis and therefore had hematogenous dissemination before her index surgery. CONCLUSION: In this series of patients undergoing surgery for malignancies on the gynecologic oncology service, blood salvage with LRF was not definitively associated with hematogenous dissemination. Further large controlled studies are needed to demonstrate the clinical safety of the use of blood salvage in this setting.

Indocyanine green dye-enhanced transpupillary thermotherapy of classic subfoveal choroidal neovascularization.

BACKGROUND AND OBJECTIVE: To evaluate the efficacy of indocyanine green dye-enhanced transpupillary thermotherapy for the treatment of classic subfoveal choroidal neovascular membrane (CNVM) with differing etiologies. PATIENTS AND METHODS: Nine eyes of 9 patients with classic subfoveal CNVM with different etiologies that was treated with indocyanine green dye-enhanced transpupillary thermotherapy were prospectively studied. All patients underwent pretreatment fluorescein angiography. Three minutes after dye injection (25 mg of indocyanine green), transpupillary thermotherapy was delivered using a modified diode laser at 810 nm, with a variable spot size of 0.8 to 2 mm depending on the size of the CNVM. A power range between 100 and 800 mW was used and treatment was initiated in one spot for 60 seconds; the end point was a barely detectable light gray appearance of the lesion. The outcome measures were assessed by Snellen visual acuity chart and obliteration of the CNVM by fluorescein angiography. Mean follow-up was 16 months. RESULTS: Obliteration of the CNVM was seen in all patients during the last visit. Visual acuity improved in one eye and remained stable (+/-2 line change on Snellen chart) in three eyes. Three eyes revealed a moderate visual loss of 3 to 4 lines and two eyes had a severe decline (> 5 lines) in vision. None of the eyes exhibited recurrence of the CNVM during follow-up. CONCLUSIONS: Indocyanine green dye-enhanced transpupillary thermotherapy was highly effective in closing classic CNVM in all cases without any recurrence. Visual acuity was either stabilized or improved in 4 of 9 eyes (44.5%).

Transpupillary thermotherapy for choroidal neovascular membrane in age related macular degeneration.

PURPOSE: To evaluate the efficacy of transpupillary thermotherapy (TTT) in choroidal neovasularisation (CNVM) secondary to age related macular degeneration (AMD). MATERIAL AND METHODS: Retrospective, non-randomized study of 28 eyes of 28 patients with subfoveal CNVM (classic, occult or mixed) secondary to AMD. RESULTS: Fifteen patients (53.57%) maintained their pre-treatment vision, 2 (7.14%) patients showed improvement of more than 2 lines and 11 (39.28%) patients showed deterioration of vision by >2 lines. Angiographic and clinical regression of CNVM was noted in 19 patients (67.8%) on an average follow up of 15.32 +/- 3.31 months. CONCLUSION: TTT leads to stabilisation of vision in 60% of treated eyes with CNVM due to AMD.

N-hydroxyethyl-piperidine and -pyrrolidine homoazasugars: preparation and evaluation of glycosidase inhibitory activity.

An efficient and practical strategy for the synthesis of N-hydroxyethyl-1-deoxy-homonojirimycins 4 and 5 and N-hydroxyethyl-pyrrolidine homoazasugars 6 and 7 with full stereocontrol is being reported. The key step involved is the intermolecular Michael addition of benzylamine to D-glucose derived alpha,beta-unsaturated ester 8 followed by N-alkylation with ethyl bromoacetate. Reduction with LAH, acetylation, hydrogenation and protection with -Cbz group afforded compounds 14a and 14b. Removal of 1,2-acetonide functionality, hydrogenation and deacetylation afforded N-hydroxyethyl-D-gluco-1-deoxyhomonojirimycin (4) and N-hydroxyethyl-L-ido-1-deoxyhomonojirimycin (5), respectively. Compounds 14a and 14b on acetylation followed by removal of 1,2-acetonide functionality, sodium metaperiodate oxidation, hydrogenation and deacetylation gave 1,4,5-trideoxy-1,4-imino-N-hydroxyethyl-D-arabino-hexitol (6) and 1,4,5-trideoxy-1,4-imino-N-hydroxyethyl-L-xylo-hexitol (7), respectively. The glycosidase inhibition activity of compounds 4, 5, 6, 7, 16a and 16b was evaluated using sweet almond seed as a rich source of different glycosidases.

Comparison of pH-adjusted bupivacaine with a mixture of non-pH-adjusted bupivacaine and lignocaine in primary vitreoretinal surgery.

PURPOSE: To compare the efficacy of alkalinized bupivacaine with that of a mixture of nonalkalinized bupivacaine and lignocaine for local anesthesia in primary vitreoretinal surgery. METHODS: Through a prospective, double-blind, randomized, controlled clinical trial, 540 consecutive patients undergoing primary vitreoretinal surgery received either alkalinized 0.5% bupivacaine (group B) or a mixture of nonalkalinized 0.5% bupivacaine and 2% lignocaine (group BL), both of which were administered with hyaluronidase, for periocular anesthetic block. The periocular block involved two injections in the extraconal space-one in the lower temporal quadrant and the other in the medial periconal space. The efficacy of the block was graded from 0 to 5 depending on the adequacy of anesthesia and akinesia and the need for local supplementation. RESULTS: Adequate anesthesia and akinesia (grade 5) were achieved in 72.2% of the patients in group B compared with 57.4% in group BL (P = 0.0003). Intraoperative supplementation was needed in 9.6% and 20.7% of the patients in groups B and BL, respectively (P = 0.0003). Postoperative analgesics were required in 7.4% of the patients in group B and in 15.2% of those in group BL (P = 0.004). CONCLUSION: Alkalinized 0.5% bupivacaine provides better quality of anesthesia than does the mixture of nonalkalinized 0.5% bupivacaine and 2% lignocaine for patients undergoing primary vitreoretinal surgery.