To evaluate the biological impacts of high background radiation exposures that are represented by... more To evaluate the biological impacts of high background radiation exposures that are represented by spontaneous deoxyribonucleic acid (DNA) damage, an evaluation on lymphocyte cells from residents of Mamuju, West Sulawesi, Indonesia was tested. The mean annual dose received by individuals in this area is about 10.40 mSv. Of the 177 adult subjects studied, 102 were from high-level natural radiation areas of Mamuju and 75 subjects were from a nearby normal-level natural radiation area. Both areas are similar in living situations. DNA strand breaks and other parameters of study and control group were determined using a standardized comet assay. Our results showed that chronic low-level natural radiation had resulted in significantly higher (p<0.001) DNA damage based on the three parameters of the assay (tail length, tail DNA, and tail moment) compared to those of control. There was a positive correlation between the level of DNA damage and age, where people aged 40 years and older had...
Radiation protection is an action or effort taken to reduce the effects of damaging radiation exp... more Radiation protection is an action or effort taken to reduce the effects of damaging radiation exposure. Therefore, it is necessary to routinely monitor the condition of workers, among others, with a whole-body counter (WBC) either for daily activities or in a radiation emergency. In this research 78 male and female workers with an average age of 44.51 years old who have been worked for 1 up to more than 30 years at the Center for Technology of Radiation Safety and Metrology (PTKMR) - BATAN in 2018-2019 have been determined with standard procedures. The results showed that the average K-40 count for the year 2018 was 3767.35 ± 975.33 Bq. Among the 80 participants tested, internal radio-isotopic contamination with Cs-137 was detected in one person, but the levels were marginal and just exceeded the detection limit (250 Bq/body). This average value was lower than that of the previous year (2017) i.e. 4274.74 ± 740.85 Bq for 43 respondents. There is no positive correlation between radio...
Malaria is a major public health problem in Indonesia. Therefore, an effective vaccine against th... more Malaria is a major public health problem in Indonesia. Therefore, an effective vaccine against this disease is actively being sought by using gamma rays to attenuate the parasites. However, the safety and efficacy of the resulting vaccine are dependent on the precise irradiation dose. The aim of this research was to determine the exact time when the parasites are attenuated by gamma ray exposure. Mice blood containing Plasmodium berghei of 5,0 X 107 parasites/ml was irradiated with gamma rays at doses of 0, 150, 175 and 200 Gy (doses rate of 380 Gy/h) and then was injected intraperitoneally to mice at 0, 1, 2, 3, and 4 h post irradiation. The parasitemia (parasite density) in mouse blood was observed starting with day 2 and repeated every 2-4 days up to 28 days. The survival of the mice was also observed during the experiment. The results showed that the pre-patent period advanced with exposing infected blood to 150 and 175 Gy irradiations, suggesting some degree of attenuation. The...
In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPB... more In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear budding (NBUD) are the most commonly analysed morphological types of nuclear abnormalities. In contrast, tailed and dumbbell-shaped nucleus have historically received little attention in the CBMN assay. Interestingly, the incidence of tailed and dumbbell-shaped nuclei in lymphocytes is closely related with that of dicentric chromosomes or NPBs in the CBMN assay. To provide a better picture of the implications and significance of tailed and dumbbell-shaped nuclei as markers of radiation exposure, a literature review was performed in this study. Twenty articles were found in PubMed, PubMed Central, and manually searched. The articles were screened and those that met the inclusion criteria and did not meet the exclusion criteria were reviewed by all authors. At the end, nine articles were included. In conclusion, the assessment of in vivo tailed nuclei in blood smears and ac...
Malaria is an important cause of severe anemia in pregnant women, and by this mechanism malaria c... more Malaria is an important cause of severe anemia in pregnant women, and by this mechanism malaria causes around 10,000 maternal deaths each year. In the world, research on the malaria vaccine in pregnancy has also been relatively neglected. In this paper, we studied the effects of gamma irradiated P. berghei injection to pregnant mouse and its progenies. On the day 9 of pregnancy, a number of 150, 175 and 200Gy irradiated parasites in blood were injected intraperitoneally to mouse. The monitoring of parasitemia in blood of pregnant and progenies were started 3 days after infection and 2 weeks after delivery, respectively. The number, body weight and survival times of progenies were recorded. Results showed that non irradiated parasites injection produced less progenies than those of irradiated parasites. There were no parasites in blood of progenies of mother injected with non irradiated and irradiated infected bloods. Peak of parasitemia and survival were similar in infected pregnant...
Telah dilakukan penandaan DNA dengan 32P untuk deteksi resistensi M.tuberculosis terhadap isoniaz... more Telah dilakukan penandaan DNA dengan 32P untuk deteksi resistensi M.tuberculosis terhadap isoniazid dengan teknik biologi molekuler berbasis nuklir. Tuberkulosis (TB) menempati urutan pertama penyebab kematian akibat penyakit infeksi di Indonesia. Salah satu penyebab sulitnya pengendalian TB adalah merebaknya M.tuberculosisyang resisten terhadap isoniazid. Dalam penelitian ini resistensi isoniazid dideteksi dengan menganalisis gen inhA yang merupakan salah satu penyandi resistensi isoniazid. Analisis dilakukan dengan teknik polymerase chain reaction (PCR) untuk mengamplifikasi deoxyribonucleic acid (DNA) dari gen inhA sekaligus dilabel dengan alfa 32P deoxy cytosin triphosphat ([α-32P]dCTP). DNA hasil amplifikasi dianalisis dengan teknik single strand conformation polymerism (SSCP) yang didasarkan pada perubahan mobilitas pita DNA dalam gel poliakrilamid setelah divisualisasi dengan autoradiografi. Hasil analisis terhadap 100 sampel yang diuji, diketahui bahwa 13,0% sampel diduga re...
Malaria infection kills more than one million human every year, mainly under-5-year-old children,... more Malaria infection kills more than one million human every year, mainly under-5-year-old children, including in South East Asian nations. Gamma radiation given at a single dose is commonly used to create the attenuated Plasmodium parasites to get vaccine materials. However, there is no study on the infectivity of parasites after fractionated γ-radiation. This study aimed to assess the infectivity of parasites after irradiated with fractionated γ-rays in mice. A number of Plasmodium bergheithat was irradiated in two fractions of 100 and 50 Gy, 100 and 75 Gy; and 100 and 100 Gy within 5 minutes of interval time was injected intraperitoneally into 12 mice. Mice injected with unirradiated parasites (0 Gy) served as a control group. The parasitemia level of intraerythrocytic parasites in each group was observed at days post injection up to 20 days by making Giemsa stained thin blood smears and observed under the microscope. Results showed that fractionation radiation did not effectively a...
The dose response of the number of micronuclei in cytokinesis-blocked lymphocytes after in-vitro ... more The dose response of the number of micronuclei in cytokinesis-blocked lymphocytes after in-vitro irradiation of whole blood with x rays in the dose range 0-4 Gy was studied for a heterogeneous population of 10 donors. One thousand binucleated cells were systematically scored for micronuclei. A linear-quadratic dose dependency of the micronucleus yields was observed. The data were used to derive the accuracy of the dose assessment with the technique in case of a radiation accident. It was shown that for doses of 2 Gy and higher, the uncertainty of the dose was predominantly due to interindividual differences in the x-ray-induced micronucleus yields. At the 95% confidence level, doses lower than 0.3 Gy could not be unequivocally detected with the cytokinesis-blocked micronucleus assay due to the variability of the baseline micronucleus frequency within the donor population and the poor statistical accuracy when scoring 1,000 cells. The precision and sensitivity of the method in the low-dose range could be improved by the knowledge of individual pre-irradiation baseline values and an increase of the number of cells scored by automation of the technique.
Gel electrophoresis is a standardized technique that is routinely used in many daily needs, such ... more Gel electrophoresis is a standardized technique that is routinely used in many daily needs, such as clinical laboratories, forensic and industry, molecular biology and biochemistry, conservational biology, and medicine. This paper reviews several types of techniques, with two mostly used technique being agarose and acrylamide gel electrophoresis, either isotopic or non-isotopic based analysis. This technique exploits the difference in size and charge of various molecules in the sample being analyzed. An example of the clinical application of gel electrophoresis is to detect the presence of quantitatively or qualitatively abnormal protein bands. Alterations in the relative concentration of sample fractions allow easy recognition of pathological disorders associated with a disease. In the food industry, it is applied to safeguard the quality of food products. The advantages of this technique are also described that are simple, sensitive, short analysis time, low cost, and environment ...
DETECTION OF Helicobacter pylori WITH PCR TECHNIQUE AND IMPLEMETATION OF P-LABELED DNA FOR STUDYI... more DETECTION OF Helicobacter pylori WITH PCR TECHNIQUE AND IMPLEMETATION OF P-LABELED DNA FOR STUDYING ITS RESISTANCE TO THE DRUG. Infection with Helicobacter pylori (H. pylori) could result in chronic gasthritis and may increases stomach cancer risk mainly H. pylori strains with the cag pathogenicity island (PAI). In this research, detection on the presence of H. pylori was done in 73 fresh gastric biopsies and 10 paraffin embedded tissues (slide) obtained from gasthritis patients who underwent endoscopic examination in RSCM Jakarta. Deoxyribonucleic acid (DNA) extract was amplified with polymerase chain reaction (PCR) for primer of colonization related genes such as cagA, ureA, ureC and 16S RNA gene. Four (5.47%) of 73 samples showed positive result for all genes examined except for ureA gene which were also positive for urease test, of which 2 other samples showed non-specific bands. It was predicted that this result was affected by the size of PCR product. No positive result for sl...
Efficient deoxyribonucleic acid (DNA) extraction is critical for a good polymerase chain reaction... more Efficient deoxyribonucleic acid (DNA) extraction is critical for a good polymerase chain reaction (PCR) performance in molecular diagnostic, genomic and epidemiological studies of malaria. Blood collected on filter paper is a pratical method for this puposes in high parasitemic individuals. In this paper we evaluated five different methods (Commercial kit, Chelex-saponin, microwave, methanol and Tris-EDTA) for DNA extraction. Blood specimens from six asymptomatic Plasmodium sp.-infected patients in Jayapura General Hospital were spotted onto filter paper, dried, and transported for processing. Evaluation of the results of DNA extraction was carried out by measuring yields using spectrophotometry and PCR amplification for mitochondrial gene. Results showed that the best results of PCR obtanied only with commercial kit method, whereas very low quality of PCR by microwave and no amplified DNA for three other methods. The purity of DNA for all methods was low. The sensitivity of detecti...
To evaluate the biological impacts of high background radiation exposures that are represented by... more To evaluate the biological impacts of high background radiation exposures that are represented by spontaneous deoxyribonucleic acid (DNA) damage, an evaluation on lymphocyte cells from residents of Mamuju, West Sulawesi, Indonesia was tested. The mean annual dose received by individuals in this area is about 10.40 mSv. Of the 177 adult subjects studied, 102 were from high-level natural radiation areas of Mamuju and 75 subjects were from a nearby normal-level natural radiation area. Both areas are similar in living situations. DNA strand breaks and other parameters of study and control group were determined using a standardized comet assay. Our results showed that chronic low-level natural radiation had resulted in significantly higher (p<0.001) DNA damage based on the three parameters of the assay (tail length, tail DNA, and tail moment) compared to those of control. There was a positive correlation between the level of DNA damage and age, where people aged 40 years and older had...
Radiation protection is an action or effort taken to reduce the effects of damaging radiation exp... more Radiation protection is an action or effort taken to reduce the effects of damaging radiation exposure. Therefore, it is necessary to routinely monitor the condition of workers, among others, with a whole-body counter (WBC) either for daily activities or in a radiation emergency. In this research 78 male and female workers with an average age of 44.51 years old who have been worked for 1 up to more than 30 years at the Center for Technology of Radiation Safety and Metrology (PTKMR) - BATAN in 2018-2019 have been determined with standard procedures. The results showed that the average K-40 count for the year 2018 was 3767.35 ± 975.33 Bq. Among the 80 participants tested, internal radio-isotopic contamination with Cs-137 was detected in one person, but the levels were marginal and just exceeded the detection limit (250 Bq/body). This average value was lower than that of the previous year (2017) i.e. 4274.74 ± 740.85 Bq for 43 respondents. There is no positive correlation between radio...
Malaria is a major public health problem in Indonesia. Therefore, an effective vaccine against th... more Malaria is a major public health problem in Indonesia. Therefore, an effective vaccine against this disease is actively being sought by using gamma rays to attenuate the parasites. However, the safety and efficacy of the resulting vaccine are dependent on the precise irradiation dose. The aim of this research was to determine the exact time when the parasites are attenuated by gamma ray exposure. Mice blood containing Plasmodium berghei of 5,0 X 107 parasites/ml was irradiated with gamma rays at doses of 0, 150, 175 and 200 Gy (doses rate of 380 Gy/h) and then was injected intraperitoneally to mice at 0, 1, 2, 3, and 4 h post irradiation. The parasitemia (parasite density) in mouse blood was observed starting with day 2 and repeated every 2-4 days up to 28 days. The survival of the mice was also observed during the experiment. The results showed that the pre-patent period advanced with exposing infected blood to 150 and 175 Gy irradiations, suggesting some degree of attenuation. The...
In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPB... more In the cytokinesis-block micronucleus (CBMN) assay, micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear budding (NBUD) are the most commonly analysed morphological types of nuclear abnormalities. In contrast, tailed and dumbbell-shaped nucleus have historically received little attention in the CBMN assay. Interestingly, the incidence of tailed and dumbbell-shaped nuclei in lymphocytes is closely related with that of dicentric chromosomes or NPBs in the CBMN assay. To provide a better picture of the implications and significance of tailed and dumbbell-shaped nuclei as markers of radiation exposure, a literature review was performed in this study. Twenty articles were found in PubMed, PubMed Central, and manually searched. The articles were screened and those that met the inclusion criteria and did not meet the exclusion criteria were reviewed by all authors. At the end, nine articles were included. In conclusion, the assessment of in vivo tailed nuclei in blood smears and ac...
Malaria is an important cause of severe anemia in pregnant women, and by this mechanism malaria c... more Malaria is an important cause of severe anemia in pregnant women, and by this mechanism malaria causes around 10,000 maternal deaths each year. In the world, research on the malaria vaccine in pregnancy has also been relatively neglected. In this paper, we studied the effects of gamma irradiated P. berghei injection to pregnant mouse and its progenies. On the day 9 of pregnancy, a number of 150, 175 and 200Gy irradiated parasites in blood were injected intraperitoneally to mouse. The monitoring of parasitemia in blood of pregnant and progenies were started 3 days after infection and 2 weeks after delivery, respectively. The number, body weight and survival times of progenies were recorded. Results showed that non irradiated parasites injection produced less progenies than those of irradiated parasites. There were no parasites in blood of progenies of mother injected with non irradiated and irradiated infected bloods. Peak of parasitemia and survival were similar in infected pregnant...
Telah dilakukan penandaan DNA dengan 32P untuk deteksi resistensi M.tuberculosis terhadap isoniaz... more Telah dilakukan penandaan DNA dengan 32P untuk deteksi resistensi M.tuberculosis terhadap isoniazid dengan teknik biologi molekuler berbasis nuklir. Tuberkulosis (TB) menempati urutan pertama penyebab kematian akibat penyakit infeksi di Indonesia. Salah satu penyebab sulitnya pengendalian TB adalah merebaknya M.tuberculosisyang resisten terhadap isoniazid. Dalam penelitian ini resistensi isoniazid dideteksi dengan menganalisis gen inhA yang merupakan salah satu penyandi resistensi isoniazid. Analisis dilakukan dengan teknik polymerase chain reaction (PCR) untuk mengamplifikasi deoxyribonucleic acid (DNA) dari gen inhA sekaligus dilabel dengan alfa 32P deoxy cytosin triphosphat ([α-32P]dCTP). DNA hasil amplifikasi dianalisis dengan teknik single strand conformation polymerism (SSCP) yang didasarkan pada perubahan mobilitas pita DNA dalam gel poliakrilamid setelah divisualisasi dengan autoradiografi. Hasil analisis terhadap 100 sampel yang diuji, diketahui bahwa 13,0% sampel diduga re...
Malaria infection kills more than one million human every year, mainly under-5-year-old children,... more Malaria infection kills more than one million human every year, mainly under-5-year-old children, including in South East Asian nations. Gamma radiation given at a single dose is commonly used to create the attenuated Plasmodium parasites to get vaccine materials. However, there is no study on the infectivity of parasites after fractionated γ-radiation. This study aimed to assess the infectivity of parasites after irradiated with fractionated γ-rays in mice. A number of Plasmodium bergheithat was irradiated in two fractions of 100 and 50 Gy, 100 and 75 Gy; and 100 and 100 Gy within 5 minutes of interval time was injected intraperitoneally into 12 mice. Mice injected with unirradiated parasites (0 Gy) served as a control group. The parasitemia level of intraerythrocytic parasites in each group was observed at days post injection up to 20 days by making Giemsa stained thin blood smears and observed under the microscope. Results showed that fractionation radiation did not effectively a...
The dose response of the number of micronuclei in cytokinesis-blocked lymphocytes after in-vitro ... more The dose response of the number of micronuclei in cytokinesis-blocked lymphocytes after in-vitro irradiation of whole blood with x rays in the dose range 0-4 Gy was studied for a heterogeneous population of 10 donors. One thousand binucleated cells were systematically scored for micronuclei. A linear-quadratic dose dependency of the micronucleus yields was observed. The data were used to derive the accuracy of the dose assessment with the technique in case of a radiation accident. It was shown that for doses of 2 Gy and higher, the uncertainty of the dose was predominantly due to interindividual differences in the x-ray-induced micronucleus yields. At the 95% confidence level, doses lower than 0.3 Gy could not be unequivocally detected with the cytokinesis-blocked micronucleus assay due to the variability of the baseline micronucleus frequency within the donor population and the poor statistical accuracy when scoring 1,000 cells. The precision and sensitivity of the method in the low-dose range could be improved by the knowledge of individual pre-irradiation baseline values and an increase of the number of cells scored by automation of the technique.
Gel electrophoresis is a standardized technique that is routinely used in many daily needs, such ... more Gel electrophoresis is a standardized technique that is routinely used in many daily needs, such as clinical laboratories, forensic and industry, molecular biology and biochemistry, conservational biology, and medicine. This paper reviews several types of techniques, with two mostly used technique being agarose and acrylamide gel electrophoresis, either isotopic or non-isotopic based analysis. This technique exploits the difference in size and charge of various molecules in the sample being analyzed. An example of the clinical application of gel electrophoresis is to detect the presence of quantitatively or qualitatively abnormal protein bands. Alterations in the relative concentration of sample fractions allow easy recognition of pathological disorders associated with a disease. In the food industry, it is applied to safeguard the quality of food products. The advantages of this technique are also described that are simple, sensitive, short analysis time, low cost, and environment ...
DETECTION OF Helicobacter pylori WITH PCR TECHNIQUE AND IMPLEMETATION OF P-LABELED DNA FOR STUDYI... more DETECTION OF Helicobacter pylori WITH PCR TECHNIQUE AND IMPLEMETATION OF P-LABELED DNA FOR STUDYING ITS RESISTANCE TO THE DRUG. Infection with Helicobacter pylori (H. pylori) could result in chronic gasthritis and may increases stomach cancer risk mainly H. pylori strains with the cag pathogenicity island (PAI). In this research, detection on the presence of H. pylori was done in 73 fresh gastric biopsies and 10 paraffin embedded tissues (slide) obtained from gasthritis patients who underwent endoscopic examination in RSCM Jakarta. Deoxyribonucleic acid (DNA) extract was amplified with polymerase chain reaction (PCR) for primer of colonization related genes such as cagA, ureA, ureC and 16S RNA gene. Four (5.47%) of 73 samples showed positive result for all genes examined except for ureA gene which were also positive for urease test, of which 2 other samples showed non-specific bands. It was predicted that this result was affected by the size of PCR product. No positive result for sl...
Efficient deoxyribonucleic acid (DNA) extraction is critical for a good polymerase chain reaction... more Efficient deoxyribonucleic acid (DNA) extraction is critical for a good polymerase chain reaction (PCR) performance in molecular diagnostic, genomic and epidemiological studies of malaria. Blood collected on filter paper is a pratical method for this puposes in high parasitemic individuals. In this paper we evaluated five different methods (Commercial kit, Chelex-saponin, microwave, methanol and Tris-EDTA) for DNA extraction. Blood specimens from six asymptomatic Plasmodium sp.-infected patients in Jayapura General Hospital were spotted onto filter paper, dried, and transported for processing. Evaluation of the results of DNA extraction was carried out by measuring yields using spectrophotometry and PCR amplification for mitochondrial gene. Results showed that the best results of PCR obtanied only with commercial kit method, whereas very low quality of PCR by microwave and no amplified DNA for three other methods. The purity of DNA for all methods was low. The sensitivity of detecti...
Uploads
Papers by Mukh Syaifudin