Introduction. Several studies have been conducted on the use of Immunoglobulin Y (IgY) technology in the fields of diagnostics and therapeutics. IgY is the avian counterpart of the mammalian immunoglobulin G (IgG) which is exclusively transferred from the hen to the yolk thus conferring passive immunization to the growing embryo. However, despite the advantages it offers over the use of mammalian immunoglobulin, IgY technology has remained underutilized. Objective. The objective of this study is to produce an IgY with activity against synthetic peptide analogs of known immunogenic epitopes of the Hepatitis B Surface Antigen (HBsAg) - a molecular marker of Hepatitis B infection. Methods. Chickens were immunized with synthetic peptide analogs of previously reported immunogenic epitopes of the S and the pre-S1 regions of the Hepatitis B surface antigen (HBsAg). IgY specific for the synthetic peptides was isolated by delipidation and salt precipitation and was further purified by affinity chromatography. Purity and molecular weights of the whole IgY molecule and its subunits were assessed and determined by SDS-PAGE. Anti-peptide activity and specificity were determined by indirect ELISA. The study was approved by the Ethical Review Board (ERB) and Technical Review Board of the Research Implementation and Development Office (RIDO), University of the Philippines Manila. Results and Conclusion. The IgY that was purified in this study had an approximate molecular weight of 165 kilodaltons. The heavy and light chains are 60 and 28 kilodaltons, respectively. The affinity purified IgY demonstrated anti-peptide antibody activity against synthetic peptide analogs of known immunogenic epitopes of the HBsAg. Specific binding against a battery of synthetic peptides also revealed that the affinity purified IgY specifically binds to the known immunogenic epitope of the HBsAg.
HEPATITIS B SURFACE ANTIGENS ; HEPATITIS B ANTIGENS ; HEPATITIS B VIRUS ; IGY

INTRODUCTION: Asthma is an IgE-mediated inflammatory response characterized by hyperresponsiveness, airway inflammation, and reversible airflow obstruction. Currently, asthma affects 12 - 22% of the population in the Philippines. Anecdotal reports showed that yerba buena (Mentha arvensis Linn.) and oregano (Coleus amboinicus Lour.) are utilized for treating asthma in the folk culture.
OBJECTIVE: The objective of this study was to determine the effect of combined Yerba Buena (Mentha arvensis Linn.) and Oregano (Coleus amboinicus Lour.) leaves extract in asthma-induced mice.
METHODS:This study investigated the anti-asthmatic activity of the aqueous and methanolic extracts of the combined herbs in asthma-induced mice using immunoglobulin E (IgE) as a parameter.
RESULTS: Aqueous- and methanol-treated mice has 50% and 60% reduction in the IgE level, respectively (p = 0.018). The extracts exhibited a significant (p = 0.001) anti-inflammatory activity in mice that further proved its effect on IgE. Moreover, lung histopathology also established the potential effect of the extract through the widening of the alveoli on treated mice.
CONCLUSION: Combined Yerba Buena and Oregano aqueous and methanol extracts may have a potential health benefit against asthma.

Background: Earlier studies reported the anti-inflammatory activity in several species of Piper, and Piper umbellatum Linn. leaves containing some phytochemicals that are potent anti-inflammatory agents. However, there was no thorough investigation on the anti-inflammatory activity of the locally grown P. umbellatum in the Philippines. Objective: The study aimed to determine the anti-inflammatory activity of Piper umbellatum leaves using in vitro and in vivo assays. Methodology: Crude extracts were obtained from P. umbellatum leaves using polar and non-polar solvents. The anti-inflammatory activities of all crude extracts were determined using the carrageenan-induced paw edema test in mice and phytochemical analysis. The crude extract with the highest activity was partially purified using column chromatography. The fractions with similar TLC profile were pooled and tested for antiinflammatory activity. COX-1 and COX-2 enzyme inhibitory activity were determined in pooled fractions that showed initial activity in animal model. Results: Among the crude extracts of P.umbellatum, the crude ethyl acetate extract exhibited a significant dose-dependent inhibition on paw edema test with doses of 500 mg/kg bw, 1,000 mg/kg bw and 1,500 mg/kg bw (p<0.05). Among the 20 pooled fractions (PF) collected from the ethyl acetate extract, PF58, PF60 and PF64 had the highest COX-2 enzyme inhibitions of 83.12 %, 84.78% and 77.47%, respectively (p<0.05). PF60 also exhibited the highest anti-inflammatory activity on paw edema with inhibitions of 62.45% at low dose (250 mg/kg bw) and 76.10 % at high dose (1,000 mg/kg bw) in mice. Conclusion The ethyl acetate extract of P. umbellatum leaves and its fraction-PF60 exhibited a significant anti-inflammatory activity in in vitro and in vivo assays and contained high amounts of total phenolic and total flavonoid.
Prostaglandin-Endoperoxide Synthases ; Carrageenan ; Inflammation

Background: Alpha-amylase and alpha-glucosidase are essential enzymes in converting food into energy inside the body. However, the proper management of these enzymes is needed to maintain normal blood glucose levels in the body. Thus, inhibiting these carbohydrate-hydrolyzing enzymes is necessary for managing hyperglycemia and alleviating Type 2 Diabetes Mellitus complications. Studies have reported that the Piper species have inhibitory properties on these digestive enzymes. However, the locally cultivated P. umbellatum has not yet been studied. Objectives: The study evaluated the inhibitory potentials of the locally grown P. umbellatum leaf extracts against α-amylase and α-gluiosidase. Methodology: The P. umbellatum leaf extracts were screened for in vitro inhibitory α-amylase and α-glucosidase tests. The most active crude extract was subjected to semi-purification and fractions were subjected to inhibitory enzymatic tests. Qualitative and quantitative phytochemical analyses were conducted. Results: Among all crude extracts, the KMB-HE exhibited the highest activities comparable to that of the acarbose standard (p>0.05). The KMB-HE showed inhibitions of 81.01%±1.66 and 89.51%±3.03 on α-amylase and α-glucosidase at 250 μg/mL, respectively. In addition, it contained the highest phenolic (195.00 ±3.60 GAE mg/gram) and flavonoid (4.55±0.06 QE mg/gram) contents that may be linked to the observed activities. The semi-purification of KMB-HE collected 24 fractions with PF10 demonstrating inhibitions of 68.07%±13.48 and 74.21%±2.22 in α-amylase and α-glucosidase, respectively. Also, the PF10 contained the highest phenolic (204.75±2.42 GAE mg/gram) and flavonoid (4.00±0.07 QE mg/gram) compounds among all the active fractions tested. The PF10 satisfied the recommended criteria for plant natural inhibitors: a moderate-to-low α-amylase and stronger α-glucosidase inhibitory activity. Conclusion The P. umbellatum exhibited good inhibitory activities on carbohydrate-hydrolyzing enzymes. These may lead to further analysis of the P. umbellatum for future development of a safer, locally available natural remedy in managing Type 2 Diabetes Mellitus.
Diabetes Mellitus, Type 2

Background: Rattan is a member of the Arecaceae family grown in the tropical or subtropical climates. These plants are used as raw materials for furniture-making. In some cases, its shoots are also used as food and may possess bioactive potentials. In this study, rattan root methanolic extracts were screened for phytochemicals and evaluated for antioxidant activities. Methodology: Six (6) methanolic extracts of selected rattan shoots were subjected to in vitro antioxidant assays, which include DPPH, FRAP, metal chelation, superoxide, and nitric oxide. Qualitative and quantitative phytochemical analyses were also employed. Results: Shoots of Calamus sp. 02 (Bangaw-Bangaw; 85.59±0.430%), C. usitatus (Talola; 82.45±2.215%), and C. spinifolius (Kurakling; 75.54±1.599%) showed the most effective DPP radical inhibition at 66.67μg/mL. Concentration-dependent reducing power (>30% scavenging at 6.64μg/mL) with no-to-low metal chelating activity was also observed in these plant food. C. sp 02 and C. usitatus demonstrated an effective scavenging activity against superoxide anions at 227.3μg/mL. Nitric oxide scavenging activity was observed in all shoots with C. merrilli (Palasan) exhibiting highest at 78.13μg/mL. Qualitative phytochemical analyses showed that rattan shoots all contained reducing sugars, phenolics, terpenoids, and quinone compounds. Quantitative phytochemical analyses revealed that C. sp. 02 (66.024±4.183mg GAE/g) and C. merrilli (1.767±0.056mg QE/g) contained the highest amounts of phenolic and flavonoids, respectively. These phytochemicals present may explain their behavior as antioxidants. Conclusion The study revealed that different rattan shoots showed different capacities to scavenge particular oxidants. Of these, C. sp. 02, C. spinifolius, and C. merrilli may be considered promising sources of natural antioxidants.
Phytochemicals ; Antioxidants ; Calamus

Background: One of the therapeutic strategies for type 2 diabetes mellitus involves suppressing postprandial hyperglycemia by inhibiting key enzymes in carbohydrate digestion, α-glucosidase and α-amylase. While such inhibitors are commercially available, some researchers have turned to plants for potentially cheaper and safer alternatives. Objectives: The study aimed to investigate the in vitro α-glucosidase and α-amylase inhibitory activities of the leaf methanolic extracts of two native Philippine plants Ficus nota Blanco Merr. and Ficus septica Burm F, as well as their effects on postprandial blood glucose levels in a mouse model. Methodology: The in vitro activities of the leaf methanolic extracts were evaluated against porcine pancreatic α-amylase and yeast αglucosidase. The most active extract was partially purified into fractions by sequential solvent partitioning and subjected to in vitro testing. Postprandial antihyperglycemic activity was then assessed in normoglycemic ICR mice. Phytochemical analysis was also performed Results: The most active extract and fraction in vitro were FS-crude and FS-HexF, respectively, having significantly more potent αglucosidase inhibitory activity than the commercial drug acarbose. FS-crude and FS-HexF exhibited strong inhibition of αglucosidase and weak inhibition of α-amylase, which is considered favorable for novel inhibitors as it is hypothesized to reduce gastrointestinal adverse effects. However, FS-crude and FS-HexF did not significantly attenuate postprandial blood glucose levels in the oral starch tolerance test. Phytochemical analysis of FS-HexF putatively identified 6-gingerol as one of the possible bioactive components. Conclusion F. septica could be a potential source of glycoside inhibitors as it showed promising in vitro inhibition of α-amylase and α-glucosidase. While it did not exhibit significant postprandial antihyperglycemic activity in this study, more robust testing is recommended to make a definitive conclusion.
Amylases ; Glucosidases ; Hypoglycemic Agents

Background: One of the causes of inflammatory arthritis is excessive production of uric acid or hyperuricemia. It is a painful disease that is treated with a commercial xanthine oxidase inhibitor to decrease uric acid synthesis. However, the treatment is associated with adverse side effects and thus, there is interest in medicinal plants that could have similar therapeutic effects with minimal side effects. There are many reported indigenous plants and trees in the Philippines that are reported to have therapeutic and bioactive compounds. One such plant is Canarium ovatum or locally called pili. This study aimed to determine the antihyperuricemic activity of the ethanolic extract of the leaves of C. ovatum. Objective: Determine the antihyperuricemic activity of the crude ethanolic extract of C. ovatum leaves and its partially purified fractions through inhibition of xanthine oxidase and its effect on the blood uric acid level of oxonate-induced hyperuricemic mice. Methodology: The crude ethanol extract from C. ovatum leaves and its partially purified fractions obtained through column chromatography were tested for their in vitro xanthine oxidase (XO) inhibitory activity by measuring spectrophotometrically the uric acid formation from xanthine as the substrate. The crude ethanol extract and the fraction with the most XO inhibitory activity were then tested for their in vivo XO inhibitory activity in oxonate-induced hyperuricemic mice by measuring their blood uric acid levels using uric acid test strips. Results: The crude ethanolic extract of C. ovatum leaves at 100ppm showed 83.62±2.05% in vitro inhibition of XO while the most active fraction showed 80.30±4.00% inhibition. Both were comparable (p>0.05) to the positive control, allopurinol, which showed 91.47±5.64% inhibition. In vivo, the crude extract and the fraction that showed the highest XO inhibitory activity at 200 mg/kg significantly (p<0.01 and p<0.05) respectively reduced the serum uric acid levels of the hyperuricemic mice one hour after induction as compared to the negative control. Moreover, their antihyperuricemic activity were not statistically significant as compared to that of allopurinol (p<0.0001). Conclusion The crude ethanolic extract of C. ovatum leaves and its most active fraction showed statistically significant in vitro xanthine oxidase inhibition and in vivo antihyperuricemic activity. The activities shown by both crude and active fraction were not statistically different from that determined for allopurinol. Therefore, further studies can be conducted to isolate the most active compound and study its pharmacokinetic properties.
Xanthine Oxidase ; Uric Acid ; Allopurinol

Antioxidants

Plants have been a major source of natural products for sustaining human health. The use of the different parts of the plant as infusions, decoctions, extracts, and powders are being employed in the treatment of different diseases in humans, plants, and animals. One property of great significance in terms of therapeutic treatments, especially with the emergence of multi-drug resistant microbes, is the antimicrobial activity. A new promising source of antimicrobials that demonstrate novel mechanisms of therapeutic strategies is low molecular weight peptides. In this study, the antimicrobial activities of Mimosa pudica crude and partially purified peptide extracts against Gram-negative Enterobacter cloacae ATCC 23355 and Enterobacter aerogenes ATCC 13048, and Gram-positive Staphylococcus epidermidis ATCC 12228 using resazurin colorimetric assay and tricine SDS-PAGE bioautography were reported. M. pudica crude and partially purified extracts exhibited antimicrobial activity against all the bacteria tested. Specifically, the peptide that was partially purified from M. pudica with a molecular weight of 5.14 kDa inhibited the growth of Enterobacter cloacae.
Antimicrobial Peptides