Khairul Osman

A study of the antioxidant effect of alpha lipoic acids on sperm quality

OBJECTIVE Assisted reproductive techniques are useful in helping infertile couples achieve successful conception. Initial studies have shown that sperm cryopreservation, one step in assisted reproduction, causes a dramatic reduction in sperm quality. This has been attributed to, among other things, free radical activities. The aim of the present study was to minimize this oxidative attack by adding an antioxidant into the sperm microenvironment. Alpha lipoic acids were selected for this purpose for their efficient free radical scavenging properties and solubility in lipid and aqueous phases. METHODS For this investigation, semen from six Boer bucks was pooled. Seminal analysis of the baseline prior to incubation of samples with different concentrations of Alpha lipoic acids (0.00625, 0.0125, 0.025, 0.05, 0.1 mmol/ml) was performed, and post-seminal analysis was conducted after a one-hour incubation. The comet assay was used to observe the effect of Alpha lipoic acids on sperm DNA integrity. Statistical analysis using an unpaired t-test with a significance level of p<0.05 was then performed. RESULTS Our results indicate that the sperm motility rate was improved after incubation with Alpha lipoic acids at a concentration of 0.02 mmol/ml. This concentration was also capable of reducing DNA damage. CONCLUSION In conclusion, Alpha lipoic acids renders cryoprotection to sperm, thereby improving sperm quality.

Lipid Lowering Effect of Antioxidant Alpha-Lipoic Acid in Experimental Atherosclerosis

Accumulating data demonstrated that hypercholesterolemia and oxidative stress play an important role in the development of atherosclerosis. In the present study, a protective activity of alpha-lipoic acid; a metabolic antioxidant in hypercholesterolemic-induced animals was investigated. Eighteen adult male New Zealand White (NZW) rabbit were segregated into three groups labelled as group N, HCD and ALA (n = 6). Group N (normal control) was fed with normal chow, the rest (HCD and ALA) were fed with 100 g/head/day of 1% cholesterol rich diet to induce hypercholesterolemia. Four point two mg/body weight of alpha lipoic acid was concomintantly supplemented to the ALA group. Drinking water was given ad-libitum. The study was designed for 10 weeks. Blood sampling was taken from the ear lobe vein at the beginning, week 5 and week 10. Plasma was prepared for lipid profile estimation and microsomal lipid peroxidation index indicated with malondialdehyde (MDA) formation. At the end of the experiment, the animals were sacrificed and the aorta were excised for intimal lesion analysis. The plasma total cholesterol (TC) and low density lipoprotein (LDL) levels were found to be significantly low in ALA group compared to that of the HCD group (p<0.05). Similarly, low level of MDA (p<0.05) in ALA group was observed compared to that of the HCD group showing a significant reduction of lipid peroxidation activity. Histomorphometric intimal lesion analysis of the aorta showing less of atheromatous plaque formation in alpha lipoic acid supplemented group (p<0.05) compared to HCD group. These findings suggested that alpha lipoic acid posses a dual lipid lowering and anti-atherosclerotic properties indicated with low plasma TC and LDL levels and reduction of athero-lesion formation in hypercholesterolemic-induced rabbits.

Embryo apoptosis identification: Oocyte grade or cleavage stage?

Apoptosis is a programed cell death that is vital for tissue homeostasis. However, embryo apoptosis had been known to be related to embryo fragmentation which should be avoided in in vitro fertilization (IVF). The purpose of this study was to evaluate the relationship of embryo apoptosis with the grade of immature oocytes and cleavage stage of in vitro produced (IVP) cattle embryos. This study consisted of 345 oocytes collected through ovary slicing. Immature oocytes were graded as A, B and C. This grading was based on cumulus cell thickness and compactness. All oocytes then underwent an in vitro maturation (IVM) procedure. An IVF was done 24 h after IVM culture. Prior to staining, stage of cleaved embryos was determined and classified as either 2, 4, 8 or >8-cell embryo stage. Apoptosis status of cleaved IVP embryos was determined by using annexin V-FITC staining technique at 48 and 72 h post insemination (hpi). Apoptosis status for each embryo was classified as either early or late. The result showed that there was no significant difference (p > 0.05) of apoptosis status among grade A, B and C embryos. All grades of oocytes showed embryo apoptosis where 1.5% late apoptosis for grade A, 4.5% and 10.4% of early and late apoptosis for grade B and grade C. Early apoptosis was not seen in grade A embryo. We also noted no significant difference (p > 0.05) of apoptosis status between 2, 4, 8 and >8-cell embryo stage. Early apoptosis was also not seen in >8-cell stage. Even though there were no differences in apoptosis expression between the three classes, the cleavage rate of grade A oocytes was significantly higher (p < 0.01) than grade B and grade C. In conclusion, the apoptosis expression in the embryo can occur regardless of the oocyte quality and the cleavage stage of the embryo produced.

Melatonin ameliorates the adverse effects of leptin on sperm

This study examined the effects of melatonin on leptin-induced changes in sperm parameters in adult rats. Five groups of Sprague-Dawley rats were treated with either leptin or leptin and melatonin or melatonin for 6 weeks. Leptin was given daily via the intraperitoneal route (60 μg kg−1 body weight) and melatonin was given in drinking water (10 mg kg−1 or 20 mg kg−1 body weight per day). Upon completion, sperm count, sperm morphology, 8-hydroxy-2-deoxyguanosine, Comet assay, TUNEL assay, gene expression profiles of antioxidant enzymes, respiratory chain reaction enzymes, DNA damage, and apoptosis genes were estimated. Data were analyzed using ANOVA. Sperm count was significantly lower whereas the fraction of sperm with abnormal morphology, the level of 8-hydroxy-2-deoxyguanosine, and sperm DNA fragmentation were significantly higher in rats treated with leptin only. Microarray analysis revealed significant upregulation of apoptosis-inducing factor, histone acetyl transferase, respiratory chain reaction enzyme, cell necrosis and DNA repair genes, and downregulation of antioxidant enzyme genes in leptin-treated rats. Real-time polymerase chain reaction showed significant decreases in glutathione peroxidase 1 expression with increases in the expression of apoptosis-inducing factor and histone acetyl transferase in leptin-treated rats. There was no change in the gene expression of caspase-3 (CASP-3). In conclusion, the adverse effects of leptin on sperm can be prevented by concurrent melatonin administration.

Statistical discrimination of black ballpoint pen inks using ultra-performance liquid chromatography with principal component analysis

The aim of this study is to propose an approach for the analysis of black ballpoint pen writing inks based on ultra-performance liquid chromatography (UPLC) combined with principal component analysis (PCA). A total of twelve varieties of black ballpoint pens available in the Malaysian market were examined by an UPLC that coupled with a photodiode array detection (PDA). Chromatograms of ink samples were extracted at 279, 370 and 400 nm. Chromatographic data obtained were subjected to PCA after normalization. Seven principal components were produced from a total of 15 raw peaks. The new set of variables was then used for running one-way ANOVA to differentiate 66 pen-pair formed from twelve varieties of black ballpoint pen. The approach proposed here has successfully differentiated all pen-pair thus achieving 100% discrimination power.

Analisis produk penguraian haba yang terhasil daripada tisu khinzir yang terdedah kepada pembakaran terbuka

Volatile products commonly known as pyrolytic products, are thermal decomposition products generated from the various fuels that are present at a fire scene due to heat exposure. When a fire scene involves human remains, the volatile species generated from the remains can be mistaken with others residues due to the presence of interfering products or ignitable liquid residues from the fire scene. Knowledge of the type of products generated from human remains in real life fire scenarios is crucial under these circumstances. Thus, this study was executed to test the robustness and validity of the pyrolytic data generated from porcine tissue under indoor laboratory burning conditions to those generated under outdoor burning conditions. Porcine bone samples were burnt under outdoor conditions until the ignition of fat occurred (temperatures exceeding 250 °C). The pyrolytic products generated were absorbed onto activated carbon tablets that were incubated in an oven for 16 hours at 80 °C and then desorbed with pentane and injected into the Gas Chromatography-Mass Spectrometry (GC-MS). The 60 pyrolytic profiles obtained were in the range of n-alkanes, n-alkenes, n-aldehydes, aromatics and nitriles, similar to those obtained from the indoor laboratory burning data but with the additional presence of n-aldehydes. Results from this study has indicated that the human pyrolytic data model generated indoors is a good representative of outdoor burning conditions and has also successfully clarified the inconsistencies in terms of the presence and absence of n-aldehydes from porcine and human pyrolytic data.

Effect of Alcohol Consumption on the Sperm DNA Integrity: A Systematic Review

There is no solid conclusion on the conventional sperm parameters in association with alcohol consumption, evaluation of sperm DNA integrity thus become a more reliable parameter. Hereby, this literature search was performed to summarize alcohol consumption on the sperm DNA integrity. A computerized database search was done through MEDLINE via Ovid (since 1946 until August 2017) and Cochrane was used. The following set of keywords: ‘alcohol consumption OR alcohol intake OR alcohol diet OR drinking alcohol OR ethanol diet’ AND ‘sperm DNA OR sperm chromatin OR sperm genome OR sperm histone OR sperm protamine’ were utilised. 24 articles were retrieved where only five studies conform to the inclusion criteria All studies demonstrated a negative effect of alcohol consumption on sperm DNA integrity, regardless of various range of alcohol doses and duration of alcohol consumption. Out of five studies reviewed, four studies were using a different approach to measure the sperm DNA damage. Hereby, this review identified a need to use a single approach of DNA damage test by having various method of alcohol administration and/or vice versa so that the extension of sperm DNA damage to alcohol consumption will have a better conclusion. On the same note, a few studies have reported the reversibility on conventional semen parameters, none has been done on the sperm DNA damage upon alcohol withdrawal. Therefore, the role of alcohol withdrawal on the reversibility of sperm DNA damage needs to be as well investigated further.

Improvement of Post-Thaw Sperm Kinematics and DNA Integrity of Cross-Bred Bovine Sperm by Incorporating DGC as Selection Method Prior to Cryopreservation

The aim of this study was to assess post-thaw sperm quality following initial sperm selection using density gradient centrifugation (DGC) prior to cryopreservation. Ejaculates from four mature Charolais cross Kedah-Kelantan bulls were collected using artificial vagina at IBVK Pahang, Malaysia. The ejaculates were aliquoted into 3 groups: non-cryopreserved group (NC); control group of cryopreserved sperm without DGC (ND) and treatment group of sperm undergoing DGC sperm selection before cryopreservation (CDGC). Prior to analysis, samples from both cryopreserved groups were thawed at 37 °C for 30 sec. All samples were analysed for kinematics parameters, viability and compromise in DNA integrity (evaluated as DNA Fragmentation Index, DFI). All kinematics parameters were analysed using computer aided sperm analysis (CASA). Results indicated significant (p < 0.05) kinematics parameter changes for all parameters of velocity (VCL, VSL, VAP) and progression (WOB, LIN, ALH and BCF). Unfortunately, changes in spermatozoa straightness were insignificant (STR) F(2, 68) = 1.004, p = 0.371. Spermatozoa viability had increased by 26.2% (p < 0.01) following the treatment. DFI revealed the treatment group recorded a significant reduction in DFI value (0.17% fragmented DNA). In conclusion, DGC sperm selection prior to cryopreservation reduced the effects of cryodamage and showed an improvement in post-thaw sperm quality, thus reducing the occurrence of asthenozoospermia in populations of frozen-thawed cross-bred bovine spermatozoa.

The Physical and Chemical Characteristic of 30 Soil Samples from Kuala Lumpur and Selangor: A Discriminative Study

The use of soil as a form of forensic trace evidence has proven to be very useful due to its versitality and high variablility at differnt locations. Often discriminated based on their physical (color, particle size, distribution) and chemical (pH, percentage of combustible organic content and soil trace metal) characteristics, the latter is commonlyemployedas a confirmatory test due to the indivualised and accurate information generated upon analysis, particularly from the analysis using ICP-MS (Inductively Coupled Plasma-Mass Spectrometry) for trace metal analysis in soil.In this study, soil samples were collected fromthirty different locations in Selangor and Kuala Lumpur and were analyzed in terms of their physical and chemical similarity and differences.Kruskal-Wallis test, employed to test thephysical (color and particle size distribution)demonstrated that there were significant differences in terms of color between soil from different sampling locations and for particle size distribution.Upon the application of the Decision Tree test that was built using C5.0 algorithm on the physical characteristics data from soil samples, 24 out of 30 samples were differentiated. In terms of chemical characteristics, Kruskal-Wallis tests on pH and soil percentage of combustible organic content showed that there were significant differences between soil from different sampling locations, with pH[X2(29,N=90)=85.849, p<0.01] and percentage of combustible organic content [X2(29,N=90)=81.825, p<0.01]. For trace metal analysis, the ICP-MS data for each soil sample was analyzed using modified equal interval categorization method and Choropleth mapping in the form of a table and results revealed that all of the samples were individualized successfully. This study proved that with the coupling of physical and chemical characteristics of soil, any soil samples can be discriminated from each other when sampled at different locations.

Development of single tubing-type canister for cryo-storage of bull semen and their effect on sperm motility and viability

The objective of this study was to evaluate the potential of using single tubing-type canister on sperm quality. Semen was collected from the Bali cattle bull by electroejaculation technique and was cryopreserved in liquid nitrogen using slow freezing cryopreservation method. Two type of canister volume was used in this study; commercial canister (342.25π x 278 mm2) and single tubing-type canister (4π x 90 mm2). Makler counting chamber and computer assisted sperm analyzer (CASA) were used to evaluate the sperm motility and viability of post-thaw sperm. Results showed that the bull sperm motility and viability at the bottom of tubing-type canister was statistically higher and significant as compared to the commercial canister (p<0.05). Significant changes were found in sperm kinetics (VCL, VAP, VSL) of tubing-type canister compared to commercial canister. No significant changes in the motility and viability of the bull sperm at the top of tubing-type canister and commercial canister. There were no significant changes in sperm progression (LIN, WOB, PROG) in both the canisters. Developed tubing-type canister in this study showed potential as an alternative to be used in bull sperm cryo-storage.