Establishing time of death has been extensively studied for the last 30 years. Parameters that have been studied included body temperature, biochemistry of rigor mortis, putrefactive changes and entomology. Despite an extensive study in these parameters it was found that all of the parameters were very much dependent on external factors like changes in surrounding temperature and activities done prior to death. To solve this problem, we decided to monitor the mechanism that occurs during death. Until now, various researches have found that during the early stage of death, heart and perfusion to the cells will stop. This will cause the cells to start the death process. The death of the cell will occurs either through apoptosis or necrosis. During apoptosis the cells will switch on and off a few proteins in a sequence. Based on this understanding, a study was conducted to determine if area ratio of apoptosis: necrosis and apoptotic p53 and Bcl-2 markers can be used as a reliable postmortem interval marker (PMI). Sampling of the study had involved 100 dead human skins with a known PMI. All samples were obtained from forensic unit of Hospital Kuala Lumpur (UFHKL). Ratio of apoptosis: necrosis areas were determined using hematoxilin and eosin staining while apoptosis p53 and Bcl-2 markers were done using an apoptosis kit. All staining were then indexed and plotted against PMI data obtained from UFHKL. Results indicated that there were no significant correlations between ratio of apoptosis: necrosis area against PMI (p = 0.144). Whereas for both apoptotic markers p53 and Bcl-2 PMI had shown a significant correlation (p < 0.000 for both results). In conclusion, we suggest that p53 and Bcl-2 parameters should be studied further since it is very likely that it could be a good indicator for PMI.
A study was conducted to determine the radioprotective effects of Citrullus vulgaris on the lymphocyte sub-membrane particularly the actin layer. A total of 30 adult male Sprague-Dawley rats were divided into three equal groups of positive control, negative control and treatment. The positive and negative control groups were force fed with 40 ml/kg body weight of normal saline while the treatment group received 40 g/kg body weight of fresh juice of C. vulgaris daily. After a week the positive control and treatment groups were irradiated with 90 rad gamma radiation. Viable lymphocytes were determined using propidium iodine and acridine orange stain and observed under a fluorescent microscope. The percentage of viable lymphocytes of the treatment group (71.0%; p = 0.03) was significantly higher than the positive control group. The results showed that C. vulgaris possessed radioprotective effects because the lymphocyte actin was not damaged. The radioprotection effects could be due to the presence of antioxidants in C. vulgaris.
Ketidaksuburan idiopati dalam kalangan lelaki telah dikaitkan dengan kesan psikostres. Walaupun begitu, hubungan langsung antara psikostres dan ketaknormalan kualiti semen masih samar. Maka, kajian ini dijalankan untuk menentukan kesan psikostres terhadap kualiti semen terutama kesan berdasarkan residu sitoplasma dan kerosakan DNA sperma. Dalam kajian ini, responden lelaki berumur antara 25-45 tahun dipilih secara rawak dalam kalangan pesakit yang mendapatkan rawatan di Pusat Kesuburan Lembaga Penduduk dan Pembangunan Keluarga Negara (LPPKN). Seramai 331 responden akhirnya telah dipilih daripada 628 responden selepas mengambil kira faktor penolakan. Setiap responden perlu menjawab borang keizinan dan soal selidik GHQ-12 bagi penentuan tahap stres sebelum pengambilan sampel semen mengikut piawaian WHO (2010). Tahap stres diukur berdasarkan keadaan semasa responden dalam tempoh 3-4 minggu sebelum kajian. Analisis semen, pewarnaan papanicolau dan asai komet neutral digunakan untuk penentuan kualiti semen dan kerosakan DNA sperma. Keputusan menunjukkan tidak terdapat hubungan yang signifikan antara psikostres dan ketaknormalan residu sitoplasma (U=895.50, p=0.08). Namun begitu, psikostres memberi kesan kepada peratus morfologi normal (U=6317.50, p<0.05) dan kerosakan DNA sperma (U=1047.00, p<0.01). Kesimpulannya, psikostres kronik boleh menjejaskan kualiti semen dan kerosakan DNA sperma serta mempengaruhi kesuburan.
Health hazard through smart phone radiation has been associated with male infertility. The suspected prime mediator is
the NOX5 enzyme. When activated, the additional pathway for free radical production will damage sperm’s DNA. However,
conclusive evidence is still lacking. Thus, this study was conducted to comprehend the detrimental effect of the radiation
towards sperm parameters by using rat as a model. Parameters measured include sperm concentration, viability, DNA
damage status and NOX5 level on sperm. This study consisted of two phases. The first phase was conducted to determine
the optimal radiation frequency emitted from the smart phone. The radiation frequencies that were evaluated were 0 MHz
(control), 4200 MHz without multiple connection mode (minimum frequency) and 9700 MHZ with multiple connection
modes (maximum frequency). Each exposure setting represented one group. Each group consists of eight rats, which
received exposure for 6 h/day for two consecutive weeks. All parameters measured showed significant differences.
Optimum frequency for significant changes to sperm parameters were identified as the minimum frequency. Second part
of the research involved the determination of optimum exposure duration. The optimal frequency obtained was used in
combination with exposure duration of 0 h (control), 2, 4 and 6 h. Each group had 8 rats and exposure was conducted
for 2 weeks. The results showed a significant difference for all parameters following 4 h of exposure. Following this,
evaluation of DNA damage status through NOX5 activity was done by using the optimum setting where 0 MHz/0 h as a
control and 4200 MHz/4 h per day for up to 2 weeks. The results showed significant differences of NOX5 fluorescent
intensity between the two groups. In conclusion, although smart phone emitted low radiation, it can decrease sperm
concentration, viability and increase
Radiotherapy causes various complications including low immunity. Past research has shown that the low immunity is due to the low amount of lymphocytes and consumption of citrullus vulgaris will alleviate this problem. Based on this a study was conducted to identify how citrullus vulgaris was able to produce radioprotection on the lymphocyte membrane. A total of 30 adult male Sprague-Dawley rats were used and divided into three equals groups of positive control, negative control and treatment. For seven days, positive control and negative control were force fed with normal saline of 40 ml/kg animal weight while the treatment group received 40 g/kg animal weight fresh juice of citrullus vulgaris daily. After a week positive control and treatment group were irradiated with 0.9 Gy gamma ray. Viable lymphocyte were determined using propidium iodine and acridine orange stain. Results clearly shows that positive control, negative and treatment group were significantly different at 34 3% , 80 2% and 71 2% respectively. SEM results shows that pores were present on the membrane of the positive control while the negative control had none. Similar results were also found on the treatment group. Based on the result it had shown that citrullus vulgaris had radioprotection properties and lymphocytes were destroyed by the formation of pores on their membrane. It is very likely that the radioprotection properties could be due to the presence of antioxidants particularly vitamin A, C and lycopene. In conclusion, citrullus vulgaris could be used as a safe radioprotection agent.
The role of heat shock protein in reproduction is widely known as a molecular chaperone in aiding and repairing protein
formation when stress occurred. The present objectives were to evaluate the effect of different thawing temperature and
time on the expression of HSP70 gene expression and the capacitation status in cryopreserved bovine spermatozoa. Briefly,
fresh ejaculates were obtained from three different adult bulls. The semen then underwent a sperm washing technique
known as Magnetic Activated Cell Sorting System (MACS) and later on, cryopreserved. The sperm- containing straws
were then thawed at five different thawing temperatures and time post-cryostorage; 20°C for 13 s, 37°C for 30 s, 40°C
for 7 s, 60°C for 6 s and 80°C for 5 s. The RNA was extracted from each of the sperm’s pellets and converted to cDNA
prior to the qPCR process. Capacitation status was then determined by means of CTC assay. The results showed that after
the process of amplification, there is a significant different of HSP70 gene expression in MACS process samples when the
thawing process was performed at 37°C for 30 s, with p<0.05. Furthermore, the CTC assay also showed that thawing at
the same temperature gave less capacitated spermatozoa with p<0.05. As a conclusion, MACS yield spermatozoa with a
better expression of HSP70 gene and less capacitated spermatozoa when thawing was done at 37°C for 30 s.
A novel electrophoretic separation system has been successfully applied for the preparation of human sperm prior to the execution of assisted reproductive techniques (ARTs). This new system is designed to overcome the generation of reactive oxygen species (ROS) through centrifugation in conventional sperm preparation. Since the previous study showed favorable outcomes in humans, this study intends to implement this new system for animal sperm preparation particularly in bull. Fresh semen from adult bulls were used. Optimization of the electrophoretic system for optimum bull sperm separation involved different strength of voltage and separation time. The voltages applied were 10V, 20V, 30V, 40V, 50V, and 60V. For each voltage applied, the system was operated for a duration of 12 min. An average of 10 μl fractionalized semen was taken out at the collection site at every 2-min interval. Every fractionated sperm was then evaluated for percentage of viability, motility, and DNA damage assessment. Result showed that electrophoresis at 20V and 6 min yielded more than 80% viable and more than 70% motile sperm population with the lowest DNA damage. In conclusion, the system was able to fractionate high quality bull sperm at 20V and 6 min.