Targeted delivery of RNAi to cancer cells using RNA-ligand displaying exosome

Exosome is an excellent vesicle for in vivo delivery of therapeutics, including RNAi and chemical drugs. The extremely high efficiency in cancer regression can partly be attributed to its fusion mechanism in delivering therapeutics to cytosol without endosome trapping. However, being composed of a lipid-bilayer membrane without specific recognition capacity for aimed-cells, the entry into nonspecific cells can lead to potential side-effects and toxicity. Applying engineering approaches for targeting-capacity to deliver therapeutics to specific cells is desirable. Techniques with chemical modification in vitro and genetic engineering in cells have been reported to decorate exosomes with targeting ligands. RNA nanoparticles have been used to harbor tumor-specific ligands displayed on exosome surface. The negative charge reduces nonspecific binding to vital cells with negatively charged lipid-membrane due to the electrostatic repulsion, thus lowering the side-effect and toxicity. In this review, we focus on the uniqueness of RNA nanoparticles for exosome surface display of chemical ligands, small peptides or RNA aptamers, for specific cancer targeting to deliver anticancer therapeutics, highlighting recent advances in targeted delivery of siRNA and miRNA that overcomes the previous RNAi delivery roadblocks. Proper understanding of exosome engineering with RNA nanotechnology promises efficient therapies for a wide range of cancer subtypes.

Clonal Propagation of Strawberry (Fragaria x ananassa Duch.) through In vitro Runner Tip Culture through Incorporation of Growth Hormones

Runner tips explants of strawberry give rise to multiple shoots when cultured on MS medium supplemented with different concentrations and combinations of BAP with KIN or NAA or GA3.The highest response of shoot multiplication was obtained on MS containing 2.5 mgl-1 BAP + 0.5 mgl-1 Kin + 0.5 mgl-1 GA3. The maximum frequency of rooting (83%) and highest number of roots (3.49) was produced in medium containing 1.0 mgl-1 IBA. The well grown rooted plantlets were acclimatized and successfully established in autoclaved vermiculate soil and as well as natural condition. Using our established protocol, it is also possible to provide large numbers of micropropagated plantlets of this cultivars to produce high quality strawberry fruit for commercial cultivation practices.

Lymphangioma circumscriptum: clinicopathological spectrum of 29 cases

To describe the clinicopathological spectrum of Lymphangioma Circumscriptum [LC]. Observational case series. Department of Pathology and Microbiology, AKUH, Karachi, from 2002 to 2012. All reported cases of LC were retrieved from medical record. Clinical and pathological features were noted. Frequency percentages were determined. There were 29 cases of LC predominantly males [62%]. The mean age was 27.17 +/- 15.5 years. The commonest sites was anal/perianal region [24%] followed by extremities [17%] and tongue, [14%]. Vulval LC was seen in 3 patients. Two cases were described on scrotum. The lesions were most commonly suspected as viral warts, mole or polyp [in anal Region]. Vesicles with erosions and bleeding and localized growth were the usual clinical presentations. Four of the patients presented with swelling since birth. All were treated with surgical excision. Microscopic examination revealed acanthotic squamous epithelium with papillomatosis. The subepithelial region had collections of lymphatic channels composed of ectatic dilated vessels with serum and inflammatory cells in their lumina. The lymphatic channels were seen in deeper layers along with lymphocytic aggregates. Lymphangioma circumscriptum is a malformation of abnormal lymphatic channels with feeding cisterns in subcutaneous tissue. It is a benign lesion usually occurring in anal/perianal region and confused with warts. Surgical excision is preferred mode of treatment

Genetic polymorphisms of GSTM1, GSTP1 and GSTT1 genes and lung cancer susceptibility in the Bangladeshi population

Objective: To verify possible associations between polymorphisms of glutathione S-transferase Mu (GSTM1), glutathione S-transferase θ (GSTT1) and glutathione S-transferase Pi (GSTP1) genes and susceptibility to lung cancer. Methods: A total of 106 lung cancer patients and 116 controls were enrolled in a case-control study. The GSTM1 and GSTT1 were analyzed using PCR while GSTP1 was analyzed using PCR-restriction fragment length polymorphism. Risk of lung cancer was estimated as odds ratio at 95% confidence interval using unconditional logistic regression models adjusting for age, sex, and tobacco use. Results: GSTM1 null and GSTT1 null genotypes did not show a significant risk for developing lung cancer. A significantly elevated lung cancer risk was associated with GSTP1 heterozygous, mutant and combined heterozygous+mutant variants of rs1695. When classified by tobacco consumption status, no association with risk of lung cancer was found in case of tobacco smokers and nonsmokers carrying null and present genotypes of GSTM1 and GSTT1. There is a three-fold (approximately) increase in the risk of lung cancer in case of both heterozygous (AG) and heterozygous+mutant homozygous (AG+GG) genotypes whereas there is an eightfold increase in risk of lung cancer in cases of GG with respect to AA genotype in smokers. Conclusions: Carrying the GSTM1 and GSTT1 null genotype is not a risk factor for lung cancer and GSTP1Ile105Val is associated with elevated risk of lung cancer.

Effect of Ethanol Extract of Coccinia grandis Lin leaf on Glucose and Cholesterol Lowering Activity.

Aims: To investigate the effect of ethanol extract of Coccinia grandis Lin (Cucurbitaceae) leaf in glucose and cholesterol lowering activity in animal model. Study Design: Extraction, glucose and cholesterol lowering activity evaluation. Place and Duration of Study: Department of Pharmacy, North South University, Dhaka between June 2012 and December 2012. Methodology: Glucose and cholesterol lowering effect of the ethanol extract of C. grandis leaf was evaluated using the alloxan-induced diabetic rat and compared the activity with diabetic control and antidiabetic drug (Glibenclamide). Ethanol extract (25mg/kg) of C. grandis and Glibenclamide were administered to normal and experimental diabetic rats for the duration of 10 days. Results: Phytochemical screening showed the presence of alkaloids, flavonoids, saponins, cardenolides and polyprenols in significant amounts. In the alloxan-induced diabetic rat model, C. grandis (25 mg/kg) significantly (p<0.05) lowered fasting blood glucose levels. C. grandis extract (25 mg/kg) also produced significant (p<0.05) total cholesterol lowering and HDL increasing (p<0.05) effects. Surprisingly, body weight was increased significantly (p<0.05) in the C. grandis treated diabetic group. Conclusion: These results suggest that the ethanol extract of C. grandis leaf possesses significant glucose and cholesterol lowering activity in animal model, thus supporting the usage of the plant in traditional medicine as an anti-diabetic medication.

Acute Postpartum Pulmonary Edema in a 32-Year-Old Woman Five Days after Cesarean Delivery.

Acute dyspnea after pregnancy is a rare presentation, and a number of important conditions may accompany it. Pulmonary embolism, amniotic fluid embolism, pneumonia, aspiration and pulmonary edema are some of the potential causes that must be considered. The percentage of pregnancies that are complicated by acute pulmonary edema has been estimated 0.08%. The most common contributing factors include the administration of tocolytic agents, underlying cardiac disease, iatrogenic fluid overload and preeclampsia. Here we report a case of 32- year-old woman of 5th postpartum day following lower uterine cesarean section with acute dyspnea from her first pregnancy who was admitted in coronary care unit with history of one episode of raised blood pressure 160/90 mm Hg and cough on 1st postoperative day. Clinical examination and relevant investigations explored that it was a case of bilateral pulmonary edema. Patient was kept in ventilator and was treated with nitroglycerine (GTN), frusemide and ACE inhibitor. After diuresis, considerable improvement was observed in her respiratory status. From the 4th day, the patient became hemodynamically stable and was weaned off the ventilator. After five days, all the biochemical parameters became normal and she had no dyspnea.

Phytochemical screening and invitro determination of antioxidant potential of methanolic extract of streospermum chelonoides.

Plant derived phytochemicals consisting of phenols and flavonoids possess antioxidant properties, eventually renders a lucrative tool to scavenge reactive oxygen species (ROS). Current study evaluates the preliminary phytochemical screening and antioxidant activity of methanolic extract of bark and leaf of Stereospermum chelonoides. Thus, various in vitro assay strategies were implemented to evaluate antioxidant potential of Stereospermum chelonoides, using DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay, ferric reducing antioxidant power (FRAP), total antioxidant capacity, determination of total phenol and flavonoid contents. Preliminary phytochemical study revealed the presence of alkaloid, carbohydrate, glycoside, flavonoid, steroid and tannin. In DPPH radical scavenging assay, the fraction showed significant antioxidant activities in the assay compared to the reference ascorbic acid in a dose dependent manner. The IC50 value of the crude methanol extract of bark and leaf was 53.99±3.25 μg/mL and 84.73±4.02 μg/mL, respectively, whereas IC50 value for the reference ascorbic acid was 14.56±0.24 μg/mL. Moreover, profound total antioxidant activity was observed for bark (309.88±1.03 mg/g equivalent to ascorbic acid) as well as leaf (147.09±1.79 mg/g equivalent to ascorbic acid) at 200 μg/mL extract concentration. Furthermore, extract showed good reducing power capability in both bark and leaf fraction. Total phenol content for the bark was 574.82 mg/g equivalent to gallic acid and for leaf was 189.86 mg/g. For bark, the total flavonoid content was found 55.82 mg/g equivalent to quercetin and for leaf it was 49.44 mg/g.

Phytochemical screenings, thrombolytic activity and antimicrobial properties of the bark extracts of Averrhoa bilimbi.

In this present study, the bark extracts of Averrhoa bilimbi were subjected to the thrombolytic activities were assessed by using human erythrocyte and the results were compared with standard streptokinase (SK). On the other hand, bark extracts of A. bilimbi revealed moderate antibacterial activity against some microorganisms used in the screening. Preliminary phytochemical investigation suggested the presence of flavonoids, saponins and alkaloids.

Effects of Antipsychotics on the Inflammatory Response System of Patients with Schizophrenia in Peripheral Blood Mononuclear Cell Cultures

OBJECTIVE: We investigated the effects of antipsychotics on immune-challenged peripheral blood mononuclear cell (PBMC) cultures. METHODS: Blood samples were collected from twelve patients with first-episode schizophrenia. The PBMCs were separated and cultures were prepared and stimulated with lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (poly[I:C]), and then separately treated with a typical antipsychotic (haloperidol) or atypical antipsychotic (clozapine, quetiapine, or risperidone). Pro-inflammatory (interferon gamma [IFN-gamma]) and anti-inflammatory (interleukin [IL]-4 and IL-10) cytokine levels were measured in the LPS- or poly(I:C)-stimulated PBMC cultures treated with antipsychotics. RESULTS: Haloperidol and quetiapine significantly increased the IL-4 levels (p<0.05) in LPS-stimulated PBMC cultures, while clozapine and quetiapine significantly enhanced the IL-4 levels (p<0.05) in poly(I:C)-stimulated PBMC cultures. Only treatment with haloperidol resulted in a significant increase in IL-10 production (p<0.05) in LPS-stimulated PBMC cultures, whereas clozapine, quetiapine, and risperidone treatment significantly increased IL-10 production (p<0.05) in poly(I:C)-stimulated PBMC cultures. All of the antipsychotics reduced the IFN-gamma level significantly (p<0.05) in LPS- and poly(I:C)-stimulated PBMC cultures. CONCLUSION: Antipsychotic treatment altered immune function by raising the levels of anti-inflammatory cytokines (IL-4 and IL-10) and suppressing the levels of pro-inflammatory cytokines (IFN-gamma).

Strategies to evaluate antioxidant and cytotoxic activity of pet ether extract of Artocarpus chama Buch. seed.

Plant derived phytochemicals consisting of phenols and flavonoids possess antioxidant properties, eventually renders a lucrative tool to scavenge reactive oxygen species (ROS). In the current study various in vitro assay strategies were implemented to evaluate antioxidant and cytotoxic potential of pet ether extract of seeds of Artocarpus chama Buch., using DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay, cupric reducing antioxidant capacity, reducing power antioxidant capacity, total antioxidant capacity, determination of total phenol and flavonoid contents and cytotoxic activity test using brine shrimp lethality bioassay. Preliminary phytochemical study revealed the presence of flavonoid and alkaloid in the extract. In DPPH radical scavenging assay, the fraction showed significant antioxidant activities in the assay compared to the reference ascorbic acid in a dose dependent manner.The IC50 value of the crude pet ether extract was 36.87 μg/mL, whereas IC50 value for the reference ascorbic acid was 14.56 μg/mL. Moreover, profound total antioxidant activity (3676.4 mg/g equivalent to ascorbic acid) was observed at 200 μg/mL extract concentration. Furthermore, extract showed good cupric reducing power and reducing power capability. In addition, significant amount of phenols and flavonoids content were obtained from the extract. The extract also displayed strong cytotoxic potential with LC50 value of 7.19 μg/mL in brine shrimp lethality bioassay. Based on these findings, it can be concluded that significant antioxidant potential as well as cytotoxic potential of pet ether extract, might be due to the attributes of high amount of phenols and flavonoids present in the extract.

Study on Polypharmacy in Patients with Cardiovascular Diseases.

We analyzed the prevalence of polypharmacy among cardiac patients in the Natioal Institute of Cardiovascular Diseases of Dhaka, Bangladesh. Polypharmacy was defined as consumption of six or more drugs at the same time. We entered the drugs that were prescribed into the Drug Interaction Checker provided by MedScape online edition. Almost 85% of cardiac patients met criteria for three types of polypharmacy (minor, moderate and serious). However, serious and moderate types of polypharmacy were not influnenced by the increase in number of disorders (polymorbidity) as well as by the total number of drugs taken. The most frequent cause and threat that is associated with polypharmacy comes primarily from the quality of drug-drug interactions and not the total number of drugs prescribed. Most of the dangerous consecuences of polypharmacy came from the interaction of Clopidogrel with either Aspirin or PPIs. Our study emphasizes the need of informing doctors more about the problem of polypharmacy. Careful and thoughtful drug prescription strategy seems to be able to eliminate most of the cases of polypharmacy even in patients who are suffering from a multiplace disorders simultaneously. The results also provide support for development of new drugs that take into account compatibility with other medication, especially in elderly people.

In vitro evaluation of antioxidant potential of Artocarpus chama Buch. fruits.

Phytochemicals possessing phenols and flavonoids are potential sources of antioxidants, which are useful to scavenge reactive oxygen species (ROS). The present study was undertaken to evaluate and compare the antioxidant potential of pet ether and methanol extracts of fruits of Artocarpus chama Buch., using DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay, cupric reducing antioxidant capacity, reducing power antioxidant capacity, determination of total phenol and flavonoid contents. Preliminary phytochemical study revealed the presence of alkaloid and flavonoid in both extracts. The fraction showed significant antioxidant activities in the assay compared to the reference ascorbic acid in a dose dependent manner. In DPPH radical scavenging assay, the IC50 value of the crude pet ether and methanol extract was 27.64 μg/mL and 39.08 μg/mL, respectively, whereas IC50 value for the reference ascorbic acid was 12.70 μg/mL. Furthermore, both extracts showed similar cupric reducing power and reducing power capability. In addition, pet ether extract contains higher amount of phenols as compared with methanol extract, and possess similar flavonoid content expressed as Gallic acid and Quercetin, respectively. Based on these findings, it can be concluded that pet ether extract of the fruits of A. chama Buch possesses significant antioxidant potential comparing with methanol extract, which may be attributed to the high amount of phenols and flavonoids present in the extract.

Control of endogenous bacterial contamination and micropropagation of a traditional table banana (Musa spp. cv. Kanthali) of Bangladesh / 生物工程学报

Shoot tips of a traditional table banana (Musa spp. cv. Kanthali) of Bangladesh were evaluated for in vitro propagation. Initial surface sterilization (with 0.1% HgCl2 for 12 minutes) of shoot tips was successful but microbial contamination (mostly bacteria) at the rhizomatous base of the explants was observed within 6-15 days after inoculation which eventually killed 85% of inoculated explants. So, for contamination free culture establishment explants were soaked in two broad spectrum antibiotics namely ampicillin and gentamicin. Cent percent contamination free cultures were established by soaking the explants in 400 mg/L ampicillin or 200 mg/L gentamicin for 1h. Antibiotic treated explants were found to be full contamination free but failed to regenerate after 3 weeks of culture. But some of them absorbed media for up to 2nd subculture and showed swelling of explants and some color changes from pale white to light/deep green. Finally, a few days after 3rd subculture, no growth of explants was observed and all treated explants eventually started to die. Among the untreated alive explants the best medium for single shoot development was MS + 4.0 mg/L BA + 0.5 mg/L KT + 15% CW and average time required for shoot development was 18-21 days. But the regeneration percentage was very low (30%). The best medium for shoot multiplication was MS + 4.0 mg/L BA + 2.0 mg/L IAA + 15% CW and only average 3-4 shoots were formed per shoot. Finally, in vitro proliferated shoots produced roots with maximum frequency (90%) in half strength of MS medium fortified with 0.5 mg/L IBA.