1.Sperm agglutinating activity of Saccharomyces cerevisiae and Candida albicans as a potential causative factor of infertility in mice (Mus musculus).
Samuel M. GO ; Arnold V. HALLARE ; Missia Avva B. RANOLA ; Arriane C. SALANGSANG
Philippine Journal of Health Research and Development 2018;22(1):55-61
BACKGROUND AND OBJECTIVE: Vaginal yeast infections in women are usually caused by Candida albicans and, to a lesser extent, by Saccharomyces cerevisiae. Studies on C. albicans have shown that it can cause sperm agglutination which can lead to lowered fertility. This study was conducted to compare the effect of S. cerevisiae and C. albicans on the fertility of ICR mouse (Mus musculus) through sperm agglutination.
METHODOLOGY: Sperm agglutinating activity was examined by mixing different concentrations of S. cerevisiae (10, 10°, and 10 CFU/mL) and C. albicans (10", 10°, and 10 CFU/mL) separately with semen from male mice of ICR strain. Determination of the effect of S. cerevisiae and C. albicans on the fertility outcome of female mice was done by intravaginal inoculation of 20 uL of 104, 106, and 108 CFU/ml of the two yeast organisms and later allowed to mate.
RESULTS AND CONCLUSION: The study showed a statistically significantly higher percent sperm agglutination by S. cerevisiae than C. albicans at 10* CFU/ml but no difference was observed at 10° and 10 CFU/ml. No significant difference was observed in the number of sperm per agglutinate between the two yeast species at a=0.05. The concentration that exhibited the highest percentage of agglutinated sperm is 10° CFU/mL for both yeast. The most frequent type of agglutination observed in S. cerevisiae is the mixed type, while head-to-head type is most frequent in C. albicans. Both yeasts were able to cause a decline in the number of births in mice starting at 10 CFU/ml. While sperm agglutination could be one of the reasons for the infertility observed in mice, there may be other processes, mechanisms, and/or activities that could contribute to such an outcome.
Sperm Agglutination ; Candida Albicans ; Saccharomyces Cerevisiae
2.Bacterial community structure of aquaculture and non-aquaculture sediments of Taal Lake (Philippines) using PCR-DGGE of 16S rDNA
Marilen P BALOLONG ; Leslie Michelle M DALMACIO ; Maria Victoria T CAILAO ; Ma. Cecilia D TRIA ; Arnold V HALLARE
Philippine Journal of Health Research and Development 2019;23(3):48-56
Background and Objective: Microorganisms, including bacteria, serve as major players in various processes affecting both the quality of aquatic sediment as well as the fate of pollutants released into such matrix. This study, evaluated the similarity in bacterial community structure between sediments collected from aquaculture and non aquaculture sites of a tropical lake. Describing and comparing the bacterial community present in each site may provide clues on the impact of aquaculture practices on aquatic ecosystems.
Methodology: Microbial DNA was extracted using PowerSoil® DNA Isolation Kit for all sediment samples. DNA isolates were used as template in the analysis of the hypervariable region of 16S rDNA through nested polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). Excised representative 16S rDNA DGGE bands were sequenced and identified through BLAST analysis.
Results: Based on the generated mean Dice similarity coefficient of 57.77%, the bacterial community structure between aquaculture and non-aquaculture sediments was highly similar but certain taxa were found unique for each site. Bacteria belonging to Proteobacteria and Firmicutes dominated the aquaculture sediments while Proteobacteria, Firmicutes, and Chloroflexi dominated the non-aquaculture sediments. Certain physicochemical parameters operating in the two sites may have influenced the shift in representative microbes. Shewanella baltica and Trichococcus sp. were found only in aquaculture sediment owing to their ability to tolerate quantities of ammonia and high organic matter from their environment.
Conclusions: This study described the applicability of 16S rDNA PCR-DGGE as a culture-independent technique for describing and comparing the similarity between bacterial communities in sediment. Based on the generated similarity index, the bacterial community between aquaculture and non-aquaculture sediments of Taal Lake was highly similar but interestingly, harbored unique bacterial populations as seen in the DGGE profiles. The shift in dominant taxa and unique representatives per site may have been influenced by certain differences between each site's physico-chemical parameters.
Aquaculture
3.Effects of 2,3,5,6-Tetramethylpyrazine on alcohol-induced injury in liver cells and on the early life stages of zebrafish (Danio rerio Hamilton, 1822)
Margaret L.C. De Guzman ; Ma.Khrizelle D.S. Trinos ; Arnold V. Hallare ; Richard Marvin R. Espallardo
Philippine Journal of Health Research and Development 2022;26(CAS Issue):1-15
Background:
Alcoholic liver disease (ALD) is a major health problem referring to the collection of liver damage caused by excessive alcohol intake. The search for effective and safe alternatives for compounds from plants to protect the liver from extensive damages and delay the progress to a disease is still a big effort done in the scientific community. 2,3,5,6-Tetramethylpyrazine (TMP) is a compound found in a Chinese herbal medicinal plant, Ligusticum chuanxiong Hort and in some other plants.
Objective:
This study was done to assess the hepatoprotective effects of TMP against ALD using
histopathological analysis of zebrafish livers subjected to different exposure groups. TMP has been mainly used for the treatment of cardio- and cerebrovascular diseases due to its antioxidant, anti-inflammatory, and antiapoptotic properties.
Methodology:
Adult male zebrafish were exposed to three TMP concentrations (40, 60, and 80 mg/L TMP) and to 1% v/v of ethanol. The dissected livers of the zebrafish were processed for fixing on glass slides using the H&E stains and were observed under the light compound microscopes for scoring. The safety of the TMP to the early life stages of the zebrafish was tested using the Zebrafish Embryotoxicity Test (ZFET).
Results:
Results showed that TMP was able to dose-dependently decrease mean scores for the four parameters diagnostic of ALD, i.e., steatosis, inflammation, cell death, and ballooning degeneration. These scores were comparable to those of the untreated group (no ethanol + no treatment) and positive control (ethanol + Hepasil DTXTM), with all groups' scores being statistically different from those of the negative control group (ethanol + no treatment) (p<0.05). Results for the ZFET showed that incidence of embryo mortality as well as teratogenic malformations of embryos exposed to TMP were significantly lower compared to the positive control group.
Conclusion
The hepatoprotective role of TMP was implied because anomalies such as cholestasis, vessel
congestion, and hemorrhage were only observed in the ethanol-treated group and not in the other groups. In the analysis of the early development of the embryos using the Zebrafish Embryotoxicity Test (ZFET), TMP was found to be non-toxic and non-teratogenic at concentrations used for liver treatment. These initial findings on TMP provided justification for its plausibility as a hepatoprotective compound against alcoholic liver diseases (ALD).
Liver Diseases, Alcoholic
4.Effects of Apis dorsata honey on the mRNA expression of selected CYP450, pro-apoptotic, and anti-apoptotic genes during induced cytotoxicity in cyclophosphamide-treated human lung carcinoma (A549) cells
Jay T. Dalet ; Jose Kenneth T. Narag ; Arnold V. Hallare ; Francisco T. Heralde
Acta Medica Philippina 2024;58(19):37-49
INTRODUCTION
One of the novel strategies in cancer treatment is the combination of conventional chemotherapeutic drugs and natural products. In a previous study, co-treatment of the anti-cancer drug cyclophosphamide (CP) with honey from giant honey bee (Apis dorsata) resulted to a dose-dependent increase in its cytotoxic effect in human lung carcinoma (A549) cells. However, the molecular mechanism of this combinatorial effect remains unknown.
OBJECTIVESIn this study, the effect of A. dorsata honey on the expression of selected CYP450 genes at the mRNA level, as well as the proapoptotic gene CASP8 and antiapoptotic gene BCL2 was investigated in CP-treated A549 cells.
METHODSMTT Assay was performed to determine the cell viability of A549 cells after treatment with CP with or without A. dorsata honey, as well as the EC50 of CP with honey thereafter. RT-qPCR was then performed to study the effect of A. dorsata honey on the expression of selected CYP450 genes as well as CASP8 and BCL2 genes in CPtreated A549 cells. LC-MS was carried out to screen for putative compounds in A. dorsata honey which may possibly have anti-cancer activity.
RESULTSHoney in the lowest concentration (0.6% v/v) most effectively enhanced the cytotoxic effect of CP. CYP2J2 and CYP1B1 indicated a 2.38-fold and 1.49-fold upregulation, respectively as compared to untreated cells. This cytotoxic effect is further enhanced by upregulation of CASP8 that is paralleled by a downregulation of BCL2. Phytosphingosine and sphinganine are honey constituents which may be linked to the increased cytotoxicity of CP observed in A549 cells.
CONCLUSIONThis study provides further knowledge on the molecular basis by which A. dorsata honey potentiates the cytotoxic effect of cyclophosphamide in A549 cells.
Cyclophosphamide ; Lc-ms ; Liquid Chromatography-mass Spectrometry ; Apoptosis
5.Embryotoxic and teratogenic effects of polyethylene microbeads using the zebrafish Danio rerio (Hamilton, 1822)
Margaret L.C. de Guzman ; Patricia Anne P. Chua ; Franceska S. Sedano ; Arnold V. Hallare ; Jay T. Dalet
Philippine Journal of Health Research and Development 2022;26(3):51-62
Background:
The escape of polyethylene microbeads from waste-water treatment facilities to aquatic habitats has been a major concern by scientific communities due to the adverse effects on aquatic organisms as well as the well-being of the marine and terrestrial ecosystems.
Objective:
This study was conducted to evaluate the embryotoxic and teratogenic effects of polyethylene microbeads on the early development of the zebrafish Danio rerio using the Fish Embryo Acute Toxicity Test (FET).
Methodology:
Sixty (60) zebrafish embryos were exposed to polyethylene microbead suspensions (PE-MBS) of 20 μg/L, 200 μg/L, and 2000 μg/L concentrations. Using FET, the toxicological endpoints (i.e., egg coagulation, lack of somite formation, non-detachment of tail, and lack of heartbeat) were observed every 24 hours until the 96th-hour exposure. Hatching of the embryo from the chorion was observed from 48-96 hpf (hours-post fertilization), and at least four parameters of teratogenicity (i.e., edema of the pericardium and yolk sac, bent axis, tail curvature, and collapsed swim bladder) was observed at 144 hpf.
Results:
Significant differences between means and variances in the embryotoxic and teratogenic effects were observed for all treatment groups in relation to the negative control (reconstituted water). The emulsifier control (0.01% Tween 80, p-value=0.9), the solvent control (1% DMSO, p-value = 0.9), and the 20 μg/L PE-MBS (p-value = 0.92) did not significantly differ with the negative control group. However, the positive control (5% ethanol, pvalue= 7.8) and 200 μg/L (p-value = 1.1), and 2000 μg/L (p-value = 1.48) of PE-MBS were significantly embryotoxic and teratogenic to the developing organism.
Conclusion
The high concentrations of PE-MBS (200 μg/L and 2000 μg/L) may induce early hatching, mortality, and malformations. Tukey Kramer post hoc test substantiated that PE-MBS toxicity is dose-dependent since embryotoxicity and teratogenicity increase at higher concentrations. Further studies should be conducted to know more about the adverse effects of polyethylene microbeads on the development, physiology, and genomics of freshwater fishes.
Zebrafish