Light sheet imaging and interactive analysis of the cardiac structure in neonatal mice.

Light-sheet microscopy (LSM) enables us to strengthen the understanding of cardiac development, injury, and regeneration in mammalian models. This emerging technique decouples laser illumination and fluorescence detection to investigate cardiac micro-structure and function with a high spatial resolution while minimizing photodamage and maximizing penetration depth. To unravel the potential of volumetric imaging in cardiac development and repair, we sought to integrate our in-house LSM, Adipo-Clear, and virtual reality (VR) with neonatal mouse hearts. We demonstrate the use of Adipo-Clear to render mouse hearts transparent, the development of our in-house LSM to capture the myocardial architecture within the intact heart, and the integration of VR to explore, measure, and assess regions of interests in an interactive manner. Collectively, we have established an innovative and holistic strategy for image acquisition and interpretation, providing an entry point to assess myocardial micro-architecture throughout the entire mammalian heart for the understanding of cardiac morphogenesis.

Scanning electron microscopy (SEM) and atomic absorption spectroscopic evaluation of Raphanus sativus L. seeds grown in Pakistan.

Raphanus sativus L. (Brassicacae) possesses numerous health benefits due to presence of a host of secondary metabolites in its various parts. The present study investigated the nutritive value of Raphanus sativus (RS) seeds and seed oil. Proximate and physico chemical analysis were carried out by official AOAC (Association of Official Analytical Chemists) and AOCS (American oil chemist society) methods. Scanning electron microscopy (SEM) together with energy dispersive X-ray spectroscopy (EDS) described the surface morphology along with atomic elemental composition of the sample. Mineral contents were evaluated by Atomic absorption spectroscopy. Moisture content was 8.67±0.08% whereas protein, crude fiber, crude fat, carbohydrates, total ash values were reported as 20.13±0.15%, 7.86±0.15%, 32.27±0.25%, 27.32±0.85%, 3.75±0.02% respectively. EDS determined carbon, oxygen, magnesium, sulfur and potassium in seeds. All physico-chemical properties varied insignificantly for the two extraction methods, except for acid value and unsaponifiable matter, which were higher for Soxhlet's extracted oil than cold pressed oil. The mineral composition revealed potassium in the highest concentrations in seeds and seed oil i.e.1660.65±69.26 ppm and 47.80 ± 7.02 ppm respectively. The study suggested that the seed and seed oil could be a potential source of naturally originated raw material for the nutritive and pharmaceutical aid.

Electron ionization gas chromatography-mass spectrometry (ei-gc-ms) analysis of extracted oil from Tribulus terrestris seeds.

The fine powdered form T. terrestris seeds, was extracted with n-hexane by soxhlet apparatus. The aim of the study was to analyze the T. terrestris seed oil (sample-A) by electron ionization Gas Chromatography-Mass Spectrometry (EI-GC-MS) using full scan method within mass range from 40-700 charge to mass ratio (m/z). Out of 102 compounds (1A-102A) 11 compounds (30A, 32A, 37A, 45A, 47A, 48A, 49A, 64A, 83A, 101A and 102A) could not be identified and 91 were identified by classical interpretation of the mass spectrum and by using NIST14 library with match factor > 95 of mass spectrums. While among the 91 identified compounds 18 were found common therefore finally 73 compounds were identified in the present EI-GC-MS analysis of sample-A.

Chemical analysis of Dalbergia sissoo (Roxb.) pod oil by (GC-MS) / GC-FID and evaluation of antioxidant potential.

Dalbergia sissoo (Roxb.) is one of the important plant species having extensive commercial and medicinal uses. The current study aims to assess the chemical constituents in pod oil of Dalbergia sissoo (Roxb.) by using two spectroscopic techniques i.e. GC-FID (Gas Chromatography Flame Ionization Detection) and GC-MS (Gas Chromatography Mass Spectroscopy). In GC-FID technique, nine fatty acids were identified with their respective composition, capric acid (1) (1.496%) lauric acid (2) (5.695%), myristic acid (3) (4.925%), palmitic acid (4) (10.130%), palmitoleic acid (5) (2.166%), stearic acid (6) (2.862%), oleic acid (7) (10.232%), linoleic acid (8) (22.350%) and behenic acid (9) (9.283%). In second technique, i.e. GC-MS, a series of hydrocarbons (10-37) along with two triterpenoids (38-39) were found in pod oil of the plant used. Important structure indices such as Iodine value and Saponification values were also determined. These findings can be helpful to understand the important medicinal and commercial aspects of seeds oil of the plant, like fuel value, degree of unsaturation and oxidative stability. Antioxidant testing (DPPH-Radical Scavenging Assay) was also performed on pods oil but no any significant activity was found.

Development and evaluation of immediate release diclofenac sodium suppositories.

The objective of present study was to develop and evaluate polyethylene glycol (PEG) based diclofenac sodium suppositories. This study used water soluble PEG bases (1000, 4000 and 6000) in different combinations to formulate suppositories, which were further subjected for their physicochemical properties evaluation such as weight variation, average melting point, content uniformity and disintegration. Dissolution test was used to perform the in vitro release rate studies of the suppositories. The suppository (P3) containing PEG-6000 (50%) and PEG-4000 (50%) exhibited rapid in vitro release rate of diclofenac sodium. Moreover, homogeneous distribution of diclofenac sodium is found in all six formulations. The in vitro release patterns of diclofenac sodium from the marketed Voltral suppository (100mg) and formulated suppositories were also compared and found in standard limits.

Vitex negundo induces an anticonvulsant effect by inhibiting voltage gated sodium channels in murine Neuro 2A cell line.

Vitex negundo (Vn) extract is famous for the treatment of neurological diseases such as migraine and epilepsy. These neurological diseases have been associated with abnormally increased influx of sodium ions into the neurons. Drugs that inhibit voltage gated sodium channels can be used as potent anti-epileptics. Till now, the effects of Vn on sodium channels have not been investigated. Therefore, we have investigated the effects of methalonic fraction of Vn extract in Murine Neuro 2A cell line. Cells were cultured in a defined medium with or without the Vn extract (100 µg/ml). Sodium currents were recorded using whole-cell patch clamp method. The data show that methanolic extract of Vn inhibited sodium currents in a dose dependent manner (IC50 =161µg/ml). Vn (100 µg/ml) shifted the steady-state inactivation curve to the left or towards the hyper polarization state. However, Vn did not show any effects on outward rectifying potassium currents. Moreover, Vn (100 µg/ml) significantly reduced the sustained repetitive (48±4.8%, P<0.01) firing from neonatal hippocampal neurons at 12 DIV. Hence, our data suggested that inhibition of sodium channels by Vn may exert pharmacological effects in reducing pain and convulsions.

Cardamom extract induces cell proliferation by increasing potassium currents in NIH3T3 cell line.

Amommum subulatum (Roxb.) or Cardamom extract is known to have anti-inflammatory and neuroprotective effects towards many gastrointestinal related problems. However, uptill now different fractions of cardamom extract on fibroblasts with respect to potassium channel activity have not been investigated. Therefore, present study investigated the effects of different fractions of cardamom extract on potassium channels in non-tumor NIH3T3 cell line. Phytochemical analysis of hydroalcoholic, n-hexane, butane and ethyl acetate fractions of cardamom extracts were purified and isolated by thin layer chromatography (TLC). 3T3 cells were cultured and incubated with hydroalcohol (1-2 µ/ml), n-hexane (1 µ/ml), butane (2 µ/ml) and ethyl acetate (1-2 µ/ml) for 5 hrs at 37°C. Modulation in potassium currents were recorded by whole-cell patch clamp method. The data showed two constituents Cineol (C10H18O) and Terpinyl acetate (C10H17OOCCH3) by TLC method. The present study shows that the constituents in n-hexane, hydro alcohol (1 µ/ml) and ethyl acetate (2 µ/ml) significantly increased (p<0.01) the potassium outward rectifying currents from NIH3T3 cells when compared to untreated controls cells. Whereas, butanol fraction (2 µ/ml) significantly decreased (p<0.01) the inward rectifying currents when compared to controls. Moreover hydroalcoholic and n-hexane fractions have increased the proliferation in 3T3 cell line. On the other hand butanol and ethyl acetate did not induce proliferation in 3T3 cells. Taken together, our data suggested that cardamom extract contains constituents that increased K+ currents, cell migration and proliferation and are involved in wound healing.

In vitro biological activity of decoction of Joshanda.

Joshanda is a polyherbal product, commonly practicing in inflammation of upper respiratory tract as tea. The present study was conducted to find out its antimicrobial, phytotoxic, leishmanicidal and cytotoxic activities. The decoction of the product showed profound activity against Gram positive tested pathogens especially S. aureus 36.5 mm zone of inhibition at 8.0 µg/ml. However, it was inactive against C. albicans. Closed correlation was observed between two methods in terms of results. It had potent phytotoxic activity (75%). However, it was devoid of any activity leishmanicidal and cytotoxic activity. Phytochemical studies of Joshanda showed the presence of various pharmacologically active groups.

Report: Comprehensive nutrients analysis of rhizomes of Polygonatum verticillatum.

The current study was undertaken to estimate the concentration of micronutrients (Zn, Fe, Pb, Cu, Ni, Cd, Cr, Co, Sb and Mn), macronutrients (Na, Ca and K) and essential life nutrients (proteins, fats, carbohydrates and ascorbic acid) along with ash, fiber and moisture contents. Atomic absorption spectrophotometer was employed for the analysis of micronutrients while flame photometry for macronutrients. For proximate analysis (proteins, fats, carbohydrates, ash, fibers and moisture), Association of Official Analytical Chemists methods (AOAC) were used and titration method for ascorbic acid determination. It is evident from the results that the crude extract and its fractions accumulate significant concentrations of both micro and macro nutrients. The significant quantities of essential life nutrients like proteins, fats, carbohydrates and ascorbic acid along with ash, fiber and moisture contents were also found in extracts. It is concluded that the extracts of rhizomes accumulated significant quantities of life indispensible nutrients and validated the ethnobotanical uses of the plant as tonic and energizer.

Evaluation of Viola betonicifolia for its nutrition value.

In the present study, Viola betonicifolia (whole plant) and solvent fractions obtained thereof were evaluated for various micronutrients such as lead, copper, chromium, iron, nickel, zinc, cadmium, cobalt and macronutrients such as sodium, potassium and calcium using atomic absorption spectrophotometer and flame photometry respectively. It was observed that cobalt and cadmium were not detectible in tested samples while remaining nutrients were present in variable concentrations. The concentrations of all metals were compared with the recommended limits for plants and the daily consumption of all the nutrients were calculated on the basis of dose (15 g/70 kg/day or 214 mg/Kg body weight) prescribed by hakims in their practice. The concentration of chromium was far beyond the recommended limits in almost all the samples. Proximate analysis was carried out in the powdered form of the plant. Viola betonicifolia appeared as a good source of life essential nutrients like fats (18.70%), proteins (15.70%), carbohydrates (21.42%), fiber (39.01%) and vitamin C (150 mg/100 g). In conclusion, Viola betonicifolia is an excellent source of various micro and macronutrients for human being and can be used safely as a nutritional supplement.

Antimicrobial activities of rhizomes of Polygonatum verticillatum: attributed to its total flavonoidal and phenolic contents.

The current study was undertaken to evaluate the rhizomes of Polygonatum verticillatum against various pathogenic bacteria and fungi. Broad spectrum antibacterial activity was demonstrated by the crude extract of the plant and its subsequent solvent fractions; predominantly against Gram-negative bacteria. MICs of the extracts against Escherchia coli, Salmonella typhi and Shigella flexeneri were in the range of 1.5-40 µg/ml, 03-06 µg/ml and 03-40 µg/ml, respectively. The only sensitive Gram-positive bacterium was Staphylococcus aureus with MICs in the range of 75-80 µg/ml. The fungicidal activity was limited to Microspoum canis and Fusarium solani and the MICs were in the range of 350-360 µg/ml and 190-290 µg/ml respectively. The various fractions of rhizomes contained significant concentration of total flavonoidal and total phenolic contents that could be responsible for the current findings.

Rate-dependent effect of verapamil on atrial refractoriness.

OBJECTIVES: The purpose of this study was to determine whether verapamil has rate-dependent effects on the atrial effective refractory period (AERP). BACKGROUND: Block of calcium current (I(Ca)) and rapid component of the delayed rectifier potassium current (I(Kr)) by verapamil is frequency-dependent. This may result in variable effects of verapamil on the AERP, depending on the rate. METHODS: The subjects of this study were 30 adults with a mean age of 45 +/- 13 years who did not have structural heart disease. In 20 subjects, the AERP was measured at basic drive cycle lengths (BDCLs) of 650 to 250 ms, in 50 ms decrements, before and after infusion of 0.1 mg/kg verapamil. The effective refractory periods (ERPs) were measured in the setting of autonomic blockade in 10 subjects and without autonomic blockade in 10 subjects. Ten subjects served as a control group and received a saline infusion instead of verapamil. RESULTS: Verapamil significantly prolonged the AERP at BDCLs of 650 to 500 ms (p < 0.01 or p < 0.05) and significantly shortened the ERP at BDCLs of 300 and 250 ms (p < 0.01). In the control group, there were no significant differences between the baseline and post-saline measurements of ERP. CONCLUSIONS: Verapamil prolongs AERP at slow rates and shortens AERP at rapid rates. These findings are consistent with a predominant effect on I(Ca) at rapid rates and a predominant effect on I(Kr) at slow rates.

Comparison of amiodarone versus ibutilide for the prevention of immediate recurrences of atrial fibrillation during pulmonary vein isolation.

During segmental ostial ablation for pulmonary vein isolation, pulmonary vein potentials are easily identified during sinus rhythm or left atrial pacing. Therefore, maintenance of atrial fibrillation (AF) during the procedure is desirable. However, cardioversion is occasionally followed by an immediate recurrence of AF. This study compared the efficacy of ibutilide and amiodarone in preventing immediate recurrences of AF in patients who underwent pulmonary vein isolation. The subjects of this study were 25 patients (mean age 56 +/- 10 years) who underwent pulmonary vein isolation for AF who had an immediate recurrence of AF within 60 seconds after 2 transthoracic cardioversions. The patients were randomized to receive an infusion of either 300 mg of amiodarone over 10 minutes or 1 mg of ibutilide over 5 minutes. Cardioversion was repeated 15 minutes after the drug infusion. If immediate recurrences of AF occurred 2 more times, the alternative study drug was administered, and cardioversion was repeated. Immediate recurrences of AF were suppressed by amiodarone in 8 of 10 patients (80%), and by ibutilide in 9 of 15 patients (60%, p = 0.4). After crossover, immediate recurrence of AF was suppressed in 2 of 6 patients (33%) by amiodarone, and in 1 of 2 patients (50%) by ibutilide (p = 0.6). Ibutilide and amiodarone, when used alone or in combination, prevented immediate recurrences of AF in 20 of 25 patients (80%). There were no adverse drug effects. Ibutilide and amiodarone were equally effective in suppressing immediate recurrences of AF. Overall, immediate recurrences of AF can be prevented by amiodarone and/or ibutilide in 80% of patients.

Pulmonary vein isolation for paroxysmal and persistent atrial fibrillation.

BACKGROUND: The pulmonary veins (PVs) have been demonstrated to often play an important role in generating atrial fibrillation (AF). The purpose of this study was to determine the safety and efficacy of segmental PV isolation in patients with paroxysmal or persistent AF. METHODS AND RESULTS: In 70 consecutive patients (mean age, 53 +/- 11 years) with paroxysmal (58) or persistent (12) AF, segmental PV isolation guided by ostial PV potentials was performed. The left superior, left inferior, and right superior PVs were targeted for isolation in all patients, and the right inferior PV was isolated in 20 patients. Among the 230 targeted PVs, 217 (94%) were completely isolated, with a mean of 6.5 +/- 4.2 minutes of radiofrequency energy applied at a maximum power setting of 35 W. A second PV isolation procedure was performed in 6 patients (9%). At 5 months of follow-up, 70% of patients with paroxysmal and 22% of patients with persistent AF were free from recurrent AF (P<0.001), and 83% of patients with paroxysmal AF were either free of symptomatic AF or had significant improvement. Among various clinical characteristics, only paroxysmal AF was an independent predictor of freedom from recurrence of AF (P<0.05). One patient developed unilateral quadrantopsia after the procedure. There were no other complications. CONCLUSIONS: With a segmental isolation approach that targets at least 3 PVs, a clinically satisfactory result can be achieved in >80% of patients with paroxysmal AF. The clinical efficacy of pulmonary vein isolation is much lower when AF is persistent than when it is paroxysmal.