Summary table of research proposals



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1) Cytotoxic Effects of Bangladeshi Medicinal Plant Extracts.


Authors: Uddin SJ, Grice ID, Tiralongo E.
“Seven methanolic extracts from L. indica, Clerodendron inerme, Cynometra ramiflora, Xylocarpus moluccensis, Argemone mexicana, Ammannia baccifera and Acrostichum aureum and four aqueous extracts from Hygrophila auriculata, Bruguiera gymnorrhiza, X. moluccensis and Aegiceras corniculatum showed low toxicity (IC(50) > 2.5 mg mL(-1)) against mouse fibroblasts but selective cytotoxicity (IC(50) 0.2-2.3 mg mL(-1)) against different cancer cell lines.”

Suggested Topic: Application and Identification of flavonoids, such as jaceosidin and eupatilin, extracted from Damong maria (Artemisia vulgaris)
1) The cytotoxicity of Damong Maria (Artemisia vulgaris) leaves against cancer cells
Authors: Bustalinio C., Ranieses R., Tetangco, E.

“From the experiment, it was concluded that damong-maria is cytotoxic on leukemic cells, but does not damage as much normal cells as chemotheraphy drugs do. It is strongly recommended that the experiment be repeated on a wider range of concentrations (around 0.5-1.0 g leaves/ mL ethanol) to derive the concentration with the same effect to leukemic cells as chemotheraphy drugs, and the use of a polar and non-toxic solvent aside from ethanol.”


2) The antiviral activities of artemisinin and artesunate.


Authors: Efferth T, Romero MR, Wolf DG, Stamminger T, Marin JJ, Marschall M.

“The bioactivity of artemisinin and its semisynthetic derivative artesunate is even broader and includes the inhibition of certain viruses, such as human cytomegalovirus and other members of the Herpesviridae family (e.g., herpes simplex virus type 1 and Epstein-Barr virus), hepatitis B virus, hepatitis C virus, and bovine viral diarrhea virus.”



3) Liposomal incorporation of Artemisia arborescens L. essential oil and in vitro antiviral activity.


Authors: Sinico C, De Logu A, Lai F, Valenti D, Manconi M, Loy G, Bonsignore L, Fadda AM.

“The antiviral activity was studied against Herpes simplex virus type 1 (HSV-1) by a quantitative tetrazolium-based colorimetric method

Antiviral assays demonstrated that the liposomal incorporation of A. arborescens essential oil enhanced its in vitro antiherpetic activity especially when vesicles were made with P90H.”


Suggested Topic: The proapoptotic effects of flovanoids Jaceosidin and Eupatilin (from Artemisia vulgaris) on HPV-positive cervical cancer cell lines.


  • Inhibitory effect of jaceosidin isolated from Artemisiaargyi on the function of E6 and E7 oncoproteins of HPV 16 (Lee HG, Yu KA, Oh WK, Baeg TW, Oh HC, Ahn JS, Jang WC, Kim JW, Lim JS, Choe YK, Yoon DY)

    • Jaceosidin inhibits binding between

      • oncoprotein E6 of HPV and p53 tumor suppression protein

      • oncoprotein E7 of HPV and Rb tumor suppression protein

    • Also inhibits HPV-16

    • Sorry, friends, I tried to look for the full-text article to check the protocol they used to investigate it but several sites needed a fee. I found another full-text article though:

  • Jaceosidin induces apoptosis in human ovary cancer cells through mitochondrial pathway (Lv W, Sheng X, Chen T, Xu Q, Xie X)

    • Tests used:

      • MTT Assay

      • Annexin V-FITC Apoptosis Detection kit + Flow Cytometry

      • JC-1 dye assay for determination of reduction in mitochondrial membrane potential during apoptosis

      • Gel Electrophoresis and Western Blot Analysis for protein visualization & peroxidase activity

    • I quote: “Overall, our study has proved an antitumour effect of jaceosidin, a flavone from Artemisia vestita Wall. Its mechanism underlying the effect mainly involves the induction of apoptosis through activating caspase-3 via mitochondrial pathway.”

  • As suggested, knowing its possible MOA may help us come up with a framework/protocol for the anti-HPV study. However, given the number of tests, this might turn out very expensive.

  • Sorry, cross-searches for eupatilin yielded 0 results.


Suggested Topic: Recombinant human erythropoietin for improved wound healing
RESEARCH QUESTION:

Can recombinant human erythropoietin improve healing of ischemic skin wounds in Sprague Dawley rats?


P: Sprague Dawley rats

I: treatment with recombinant human erythropoeitin

C: placebo (saline solution; albumin, sodium citrate, NaCl, citric acid, H20)

O: improved healing of ischemic skin wounds

M: case control series, histopathologic tests, collagen content test, growth factors, etc.

Literature Search:


RELEVANT ARTICLES FOUND: followed through the last 4 searches (#17, 18, 19, 20)


  • Phrase in RED is the sense of the article in a nutshell

Brines M, Cerami A. Erythropoietin-mediated tissue protection: reducing collateral damage from the primary injury response. J Intern Med. 2008 Nov;264(5):405-32.



  • Quote: “Acutely, EPO prevents programmed cell death and reduces the development of secondary, pro-inflammatory cytokine-induced injury. Within a longer time frame, EPO provides trophic support to enable regeneration and healing. As the region immediately surrounding damage is typically relatively deficient in endogenous EPO, administration of recombinant EPO can provide increased tissue protection.”

  • Quote: “EPO also mediates other effects directed towards optimizing oxygen delivery to tissues, e.g. modulating regional blood flow and reducing blood loss by promoting thrombosis within damaged vessels.”

Arcasoy MO. The non-haematopoietic biological effects of erythropoietin. Br J Haematol. 2008 Apr;141(1):14-31.



  • Quote: “A series of recent studies suggest that endogenous Epo-EpoR signalling contributes to wound healing responses, physiological and pathological angiogenesis, and the body's innate response to injury in the brain and heart.”

Sayan H, Ozacmak VH, Guven A, Aktas RG, Ozacmak ID. Erythropoietin stimulates wound healing and angiogenesis in mice. J Invest Surg. 2006 May-Jun;19(3):163-73.



  • States that the treatment increased significantly the wound breaking strength of the skin in 7 days, and also an increase in vascular endothelial growth factor.

Galeano M, Altavilla D, Bitto A, Minutoli L, Calò M, Lo Cascio P, Polito F, Giugliano G, Squadrito G, Mioni C, Giuliani D, Venuti FS, Squadrito F. Recombinant human erythropoietin improves angiogenesis and wound healing in experimental burn wounds. Crit Care Med. 2006 Apr;34(4):1139-46.


Buemi M, Galeano M, Sturiale A, Ientile R, Crisafulli C, Parisi A, Catania M, Calapai G, Impalà P, Aloisi C, Squadrito F, Altavilla D, Bitto A, Tuccari G, Frisina N. Recombinant human erythropoietin stimulates angiogenesis and healing of ischemic skin wounds. Shock. 2004 Aug;22(2):169-73.



  • Treatment with erythropoietin increased vascular endothelial growth factor levels and microvessel density in ischemic wounds, and also increased collagen content in both incisional and ischemic wounds.

Buemi M, Vaccaro M, Sturiale A, Galeano MR, Sansotta C, Cavallari V, Floccari F, D'Amico D, Torre V, Calapai G, Frisina N, Guarneri F, Vermiglio G. Recombinant human erythropoietin influences revascularization and healing in a rat model of random ischaemic flaps. Acta Derm Venereol. 2002;82(6):411-7.



  • Quote: “Findings suggest that recombinant human erythropoietin administration can improve the wound healing process, in both early and late stages of injury, by reducing inflammatory response, increasing the density of capillaries in ischaemic flaps and allowing earlier repair of a damaged area.”



TOPIC ASSESSMENT:
Topic Itself: not original topic, may have to add something to make it more innovative

Medical Significance: significant enough because of 1) search for substances to speed up wound healing and 2) will greatly help any tertiary hospital like PGH

Feasibility: moderate timeframe (about 1-2 weeks treatment, 1 week measurement and analysis), must also have skills required in inducing wounds and administering treatment

Data Analysis: histopathology analysis of collagen, measured variables are straightforward and objective measurements

Disadvantages: getting EPO may be expensive, as well as assays for growth factors; skills training in personnel as mentioned above

Suggested topic: Anti-fungal and anti-microbial properties of Lankauas

Anti-fungal property of Lankauas (Languas pyramidata)
Introduction

“Lankauas is found in Rizal, Laguna, Camarines, and Sorsogon Provinces in Luzon; in Panay; in Leyte; in Lanao and Agusan Provinces in Mindanao; and in Palawan, generally in old clearings, etc. It is sometimes planted. It also occurs in the Himalayan region through Malaya to the Moluccas.

The rhizomes are strongly aromatic. The stems are 1 to 2 meters high, and about 2.5 centimeters in diameter. The leaves are pale-green with short hairs beneath. The pyramid-like inflorescence is terminal, erect, and about 30 centimeters long. The rachis and pedicles are yellowish green, and the succulent flowers are cream-colored.
Commercial use

In Java, according to Ochse, this plant is frequently cultivated. The young rhizomes and the tender, undeveloped shoots are eaten as are also the flower buds and flowers. The rhizome is used as a condiment. Its flavor is similar to ginger, but much less pungent. It is also cooked with the sap of sugar cane or with honey and water to produce an intoxicating beverage.” (Bureau of Plant Industry http://www.bpi.da.gov.ph/Publications/mp/html/l/lankauas.htm)

Lankauas root oil is also used as perfume and resinoid (Traubaud, 1964*), and as pest repellant (OGPRP, 2008).
Medical use

Lankauas root oil, prepared from fresh and dried rhizomes of the plant (De Pooter, 1985*), has high biologic activity (Lam and Zheng, 1991*).

The possible chemoprotective property of Lankauas was studied by Zheng, Kenny, and Lam* in 1993. Two compounds fractionated from lankauas root oil, namely ethyl trans-cinnamate and ethyl 4-methoxy-trans-cinnamate, were able to induce an increase in glutathione S-transferase (GST) activity in rat liver and intestine, an important mechanism in detoxifying carcinogens (Zheng, Kenny, Lam, 1993).

Crude extracts from lankauas rhizome failed to show anti-microbial property (Cereno, Monsalud, Hilbero, 2001).

“In the Philippines the juice of the rhizome is painted on an-an, or pano blanco, a kind of skin disease. According to Guerrero [FIND THIS STUDY] the rhizomes are carminative and stimulative, and a decoction of the leaves is used for anti-rheumatic and stimulant baths.” (Bureau of Plant Industry http://www.bpi.da.gov.ph/Publications/mp/html/l/lankauas.htm)

Though lankauas is used in treating an-an (tinea corporis), there are no studies regarding the potential anti-fungal property of lankauas yet.


Possible dilemmas

Another member of the Zingiberaceae family is more popular because of its medicinal value, including anti-fungal properties. The anti-microbial and anti-fungal properties of Curcuma zedoaria (BPI link) or “luya-luyahan” was studies here. Other studies include luya-luyahan’s analgesic property, inhibitory effect on NO synthesis of activated macrophages [A Curcuminoid and Two Sesquiterpenoids from Curcuma zedoaria as Inhibitors of Nitric Oxide Synthesis in Activated Macrophages. Arch Pharm Res Vol 27, No 12, 1220-1225, 2004 (4)], and anti-tumor, genotoxicity, and anticlastogenic activities.


We must be sure that our specimen is Languas and not Curcuma.
Suggested Topic: Anti-inflammatory property of Moringa oleifera in rats with acute inflammation.


  • Protective Effect of Ethanolic Extract of Seeds of Moringa oleifera Lam. Against Inflammation Associated with Development of Arthritis in Rats (Mahajan SG, Mali RG, Mehta AA)

    • Protocol: (It covers things we learned in class!  But it’s toxic.)

      • Serum level estimation of Rheumatoid Factor (RF) value and levels of selected cytokines (TNFalpha, IL-1, and IL-6)  PERFECT! Aren’t these the cytokines Dr. Penserga likes? 

      • Also checked ESR via whole blood

      • Liver homogenate for assessment of oxidative stress

      • Histopathology to measure degree of inflammation in synovial joint

    • Study concluded that Moringa oleifera possesses promising antiarthritic property (We’ll have to refine the topic if we want to pursue this. They covered a lot of ground here, I think. Maybe we can use a different plant part or focus on a specific aspect of inflammation from those listed above?)

  • Rare dipeptide and urea derivatives from roots of Moringa oleifera as potential anti-inflammatory and antinociceptive agents. (Sashidhara KV, Rosaiah JN, Tyagi E, Shukla R, Raghubir R, Rajendran SM)

    • Inhibits TNF-a and IL-2

    • Shows significant analgesic properties (possible research expansion again?)

    • Study says that M. oleifera was/is used to control activated mast cells so further studies would be more for elucidating MOA

  • Pharmacologic properties of Moringa oleifera. 2: Screening for antispasmodic, antiinflammatory and diuretic activity (Cáceres A, Saravia A, Rizzo S, Zabala L, De Leon E, Nave F)

    • Used hot water infusions of flowers, leaves, roots, seeds and stalks or bark

      • seed infusion - significant inhibition of acetylcholine-induced contraction with an ED50 of 65.6 mg/ml bath concentration, inhibition of carrageenan-induced edema at 1000 mg/kg and diuretic activity at 1000 mg/kg

      • some activity in the roots

    • Included this just to show that if we pursue the topic, it might be best to use the seeds. Roots secondarily, though the previous study by Sasdihara et al. used roots.)


Suggested Topic: Topical Epicatechin as Alternative Treatment to Ischemic Wound Healing


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