Abstract: The estimation of accumulated radiation doses in people professionally exposed to ionizing radiation was performed using methods of biological (chromosomal aberrations frequency in lymphocytes) and physical (radionuclides analysis in urine, whole-body radiation meter, individual thermoluminescent dosimeters) dosimetry. A group of 84 "A" category employees after their work in the territory of former Semipalatinsk test site (Kazakhstan) was investigated. The dose rate in some funnels exceeds 40 μSv/h. After radionuclides determination in urine using radiochemical and WBC methods, it was shown that the total effective dose of personnel internal exposure did not exceed 0.2 mSv/year, while an acceptable dose limit for staff is 20 mSv/year. The range of external radiation doses measured with individual thermo-luminescent dosimeters was 0.3-1.406 µSv. The cytogenetic examination showed that chromosomal aberrations frequency in staff was 4.27±0.22%, which is significantly higher than at the people from non-polluting settlement Tausugur (0.87±0.1%) (р ≤ 0.01) and citizens of Almaty (1.6±0.12%) (р≤ 0.01). Chromosomal type aberrations accounted for 2.32±0.16%, 0.27±0.06% of which were dicentrics and centric rings. The cytogenetic analysis of different types group radiosensitivity among «professionals» (age, sex, ethnic group, epidemiological data) revealed no significant differences between the compared values. Using various techniques by frequency of dicentrics and centric rings, the average cumulative radiation dose for group was calculated, and that was 0.084-0.143 Gy. To perform comparative individual dosimetry using physical and biological methods of dose assessment, calibration curves (including own ones) and regression equations based on general frequency of chromosomal aberrations obtained after irradiation of blood samples by gamma-radiation with the dose rate of 0,1 Gy/min were used. Herewith, on the assumption of individual variation of chromosomal aberrations frequency (1–10%), the accumulated dose of radiation varied 0-0.3 Gy. The main problem in the interpretation of individual dosimetry results is reduced to different reaction of the objects to irradiation - radiosensitivity, which dictates the need of quantitative definition of this individual reaction and its consideration in the calculation of the received radiation dose. The entire examined contingent was assigned to a group based on the received dose and detected cytogenetic aberrations. Radiosensitive individuals, at the lowest received dose in a year, showed the highest frequency of chromosomal aberrations (5.72%). In opposite, radioresistant individuals showed the lowest frequency of chromosomal aberrations (2.8%). The cohort correlation according to the criterion of radio-sensitivity in our research was distributed as follows: radio-sensitive (26.2%) — medium radio-sensitivity (57.1%), radioresistant (16.7%). Herewith, the dispersion for radioresistant individuals is 2.3; for the group with medium radio-sensitivity — 3.3; and for radio-sensitive group — 9. These data indicate the highest variation of characteristic (reactions to radiation effect) in the group of radio-sensitive individuals. People with medium radio-sensitivity show significant long-term correlation (0.66; n=48, β ≥ 0.999) between the values of doses defined according to the results of cytogenetic analysis and dose of external radiation obtained with the help of thermoluminescent dosimeters. Mathematical models based on the type of violation of the radiation dose according to the professionals radiosensitivity level were offered.
Abstract: This study was conducted to investigate the effects of brewer spent grain (BSG) on growth performance and serum biochemistry characteristics of blood of broilers chickens. Three hundred and fifteen (4 weeks old) Oba – Marshall Broilers were used for the experiment. Five experimental diets were formulated with diet 1 (T1) containing 100% soya bean meal as the control, Diet 2, 3, 4 and 5 had BSG as replacement for soya bean meal at 0%, 36%, 57%, 76% and 100% respectively. The birds were allocated into each dietary group in a completely randomized design with 63 chicks in 3 replicates of 21 chicks each. The birds were offered these diets ad libitum from four weeks old to nine weeks old (35 days). Feed intake, body weight, weight gain, and feed conversion ratio (FCR) were assessed. Blood samples were also collected to examine the effect of BSG waste on hematology and serum biochemistry of broilers. Result indicated that BSG did not significantly (P>0.05) affect feed intake and weight gain. However, FCR and final weight of finishing broilers differs significantly (P
Abstract: 10 clinically healthy hemal nodes were collected from male bulls aged 2-3 years. Light microscopy revealed a capsule of connective tissue consisted mainly of collagen fiber surrounding hemal node, numerous erythrocytes were found in wide subcapsular sinus under the capsule. The parenchyma of the hemal node was divided into cortex and medulla. Diffused lymphocytes, and lymphoid follicles, having germinal centers were the main components of the cortex, while in the medulla there was wide medullary sinus, diffused lymphocytes and few lymphoid nodules. The area occupied with lymph nodules was larger than that occupied with non-nodular structure of lymphoid cords and blood sinusoids. Electron microscopy revealed the cellular components of hemal node including elements of circulating erythrocytes intermingled with lymphocytes, plasma cells, mast cells, reticular cells, macrophages, megakaryocytes and endothelial cells lining the blood sinuses. The lymphocytes were somewhat triangular in shape with cytoplasmic processes extending between adjacent erythrocytes. Nuclei were triangular to oval in shape, lightly stained with clear nuclear membrane indentation and clear nucleoli. The reticular cells were elongated in shape with cytoplasmic processes extending between adjacent lymphocytes, rough endoplasmic reticulum, ribosomes and few lysosomes were seen in their cytoplasm. Nucleus was elongated in shape with less condensed chromatin. Plasma cells were oval to irregular in shape with numerous dilated rough endoplasmic reticulum containing electron lucent material occupying the whole cytoplasm and few mitochondria were found. Nuclei were centrally located and oval in shape with heterochromatin emarginated and often clumped near the nuclear membrane. Occasionally megakaryocytes and mast cells were seen among lymphocytes. Megakaryocytes had multilobulated nucleus and free ribosomes often appearing as small aggregates in their cytoplasm, while mast cell had their characteristic electron dense granule in the cytoplasm, few electron lucent granules were found also, we conclude that, the main function of the hemal node of cattle is proliferation of lymphocytes. No role for plasma cell in erythrophagocytosis could be suggested.
Abstract: Sodium formate is the chemical substance used for
food additive. Catalase is the important antioxidative enzyme in
protecting the cell from oxidative damage by reactive oxygen species
(ROS). The resultant level of oxidative stress in sodium formatetreated
lymphocytes was investigated. The sodium formate
concentrations of 0.05, 0.1, 0.2, 0.4 and 0.6 mg/mL were treated in
human lymphocytes for 12 hours. After 12 treated hours, catalase
activity change was measured in sodium formate-treated
lymphocytes. The results showed that the sodium formate
concentrations of 0.4 and 0.6 mg/mL significantly decreased catalase
activities in lymphocytes (P < 0.05). The change of catalase activity
in sodium formate-treated lymphocytes may be the oxidative damage
marker for detect sodium formate exposure in human.
Abstract: Azadirachta excelsa or locally known as sentang are
frequently used as a traditional medicine by diabetes patients in
Malaysia. However, less attention has been given to their toxicity
effect. Thus, the study is an attempt to examine the protective effect
of A. excelsa on the pancreas and to determine possible toxicity
mediated by the extract. Diabetes was induced experimentally in rats
by high-fat-diet for 16 weeks followed by intraperitoneal injection of
streptozotocin at dosage of 35 mg/kg of body weight. Declination of
the fasting blood glucose level was observed after continuous
administration of A. excelsa for 14 days twice daily. This is due to the
refining structure of the pancreas. However, surprisingly, the plant
extract reduced the leukocytes, erythrocytes, hemoglobin, MCHC and
lymphocytes. In addition, the rat treated with the plant extract
exhibited increment in AST and eosinocytes level. Overall, the
finding shows that A. excelsa possesses antidiabetic activity by
improving the structure of pancreatic islet of Langerhans but
involved in ameliorating of hematology and biochemical parameters.
Abstract: Cytogenetic analysis still remains the gold standard method for prenatal diagnosis of trisomy 21 (Down syndrome, DS). Nevertheless, the conventional cytogenetic analysis needs live cultured cells and is too time-consuming for clinical application. In contrast, molecular methods such as FISH, QF-PCR, MLPA and quantitative Real-time PCR are rapid assays with results available in 24h. In the present study, we have successfully used a novel MGB TaqMan probe-based real time PCR assay for rapid diagnosis of trisomy 21 status in Down syndrome samples. We have also compared the results of this molecular method with corresponding results obtained by the cytogenetic analysis. Blood samples obtained from DS patients (n=25) and normal controls (n=20) were tested by quantitative Real-time PCR in parallel to standard G-banding analysis. Genomic DNA was extracted from peripheral blood lymphocytes. A high precision TaqMan probe quantitative Real-time PCR assay was developed to determine the gene dosage of DSCAM (target gene on 21q22.2) relative to PMP22 (reference gene on 17p11.2). The DSCAM/PMP22 ratio was calculated according to the formula; ratio=2 -ΔΔCT. The quantitative Real-time PCR was able to distinguish between trisomy 21 samples and normal controls with the gene ratios of 1.49±0.13 and 1.03±0.04 respectively (p value
Abstract: Atherosclerosis was identified as a chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components (monocyte, macrophages, T lymphocytes, endothelial cells and smooth muscle cells) and the extracellular matrix of the arterial wall. Several types of genes were known to express during formation of atherosclerosis. This study is carried out to identify unknown differentially expressed gene (DEG) in atherogenesis. Rabbit’s aorta tissues were stained by H&E for histomorphology. GeneFishing™ PCR analysis was performed from total RNA extracted from the aorta tissues. The DNA fragment from DEG was cloned, sequenced and validated by Real-time PCR. Histomorphology showed intimal thickening in the aorta. DEG detected from ACP-41 was identified as cathepsin B gene and showed upregulation at week-8 and week-12 of atherogenesis. Therefore, ACP-based GeneFishing™ PCR facilitated identification of cathepsin B gene which was differentially expressed during development of atherosclerosis.
Abstract: Kwashiorkor is one of nutritional problem in
Indonesia, which lead to decrease immune system. This condition
causes susceptibility to infectious disease, especially tuberculosis.
Development of new tuberculosis vaccine will be an important
strategy to eliminate tuberculosis in kwashiorkor. Previous research
showed that 38-kDa Mycobacterium tuberculosis protein is one of the
potent immunogen. However, the role of oral immunization with 38-
kDa Mycobacterium tuberculosis protein to the number of
lymphocytes in the rat model of kwashiorkor is still unknown. We
used kwashiorkor rat model groups with 4% and 2% low protein diet.
Oral immunization with 38-kDa Mycobacterium tuberculosis protein
given with 2 booster every week. The lymphocytes number were
measured by flowcytometry. There was no significant difference
between the number of lymphocytes in the normal rat group and the
kwashiorkor rat groups. It may reveal the role of 38-kDa
Mycobacterium tuberculosis protein as a potent immunogen that can
increase the lymphocytes number from kwashiorkor rat model same
as normal rat.
Abstract: Oxidative stress is considered to be the cause for onset
and the progression of type 2 diabetes mellitus (T2DM) and
complications including neuropathy. It is a deleterious process that
can be an important mediator of damage to cell structures: protein,
lipids and DNA. Data suggest that in patients with diabetes and
diabetic neuropathy DNA repair is impaired, which prevents effective
removal of lesions. Objective: The aim of our study was to evaluate
the association of the hOGG1 (326 Ser/Cys) and XRCC1 (194
Arg/Trp, 399 Arg/Gln) gene polymorphisms whose protein is
involved in the BER pathway with DNA repair efficiency in patients
with diabetes type 2 and diabetic neuropathy compared to the healthy
subjects. Genotypes were determined by PCR-RFLP analysis in 385
subjects, including 117 with type 2 diabetes, 56 with diabetic
neuropathy and 212 with normal glucose metabolism. The
polymorphisms studied include codon 326 of hOGG1 and 194, 399
of XRCC1 in the base excision repair (BER) genes. Comet assay was
carried out using peripheral blood lymphocytes from the patients and
controls. This test enabled the evaluation of DNA damage in cells
exposed to hydrogen peroxide alone and in the combination with the
endonuclease III (Nth). The results of the analysis of polymorphism
were statistically examination by calculating the odds ratio (OR) and
their 95% confidence intervals (95% CI) using the ¤ç2-tests. Our data
indicate that patients with diabetes mellitus type 2 (including those
with neuropathy) had higher frequencies of the XRCC1 399Arg/Gln
polymorphism in homozygote (GG) (OR: 1.85 [95% CI: 1.07-3.22],
P=0.3) and also increased frequency of 399Gln (G) allele (OR: 1.38
[95% CI: 1.03-1.83], P=0.3). No relation to other polymorphisms
with increased risk of diabetes or diabetic neuropathy. In T2DM
patients complicated by neuropathy, there was less efficient repair of
oxidative DNA damage induced by hydrogen peroxide in both the
presence and absence of the Nth enzyme. The results of our study
suggest that the XRCC1 399 Arg/Gln polymorphism is a significant
risk factor of T2DM in Polish population. Obtained data suggest a
decreased efficiency of DNA repair in cells from patients with
diabetes and neuropathy may be associated with oxidative stress.
Additionally, patients with neuropathy are characterized by even
greater sensitivity to oxidative damage than patients with diabetes,
which suggests participation of free radicals in the pathogenesis of
neuropathy.
Abstract: Formaldehyde is the illegal chemical substance used
for food preservation in fish and vegetable. It can promote
carcinogenesis. Superoxide dismutases are the important
antioxidative enzymes that catalyze the dismutation of superoxide
anion into oxygen and hydrogen peroxide. The resultant level of
oxidative stress in formaldehyde-treated lymphocytes was
investigated. The formaldehyde concentrations of 0, 20, 40, 60, 80
and 120μmol/L were treated in human lymphocytes for 12 hours.
After 12 treated hours, the superoxide dismutase activity change was
measured in formaldehyde-treated lymphocytes. The results showed
that the formaldehyde concentrations of 60, 80 and 120μmol/L
significantly decreased superoxide dismutase activities in
lymphocytes (P < 0.05). The change of superoxide dismutase
activity in formaldehyde-treated lymphocytes may be the biomarker
for detect cellular injury, such as damage to DNA, due to
formaldehyde exposure.
Abstract: Fip-gts, an immunomodulatory protein purified from Ganoderma tsugae, has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. For medicinal application, a recombinant Fip-gts was successfully expressed and purified in Sf21 insect cells by our previously work. It is important to evaluate the immunomodulatory activity of the rFip-gts. To assess the immunomodulatory potential of rFip-gts, the T lymphocytes of murine splenocytes were used in the present study. Results revealed that rFip-gts induced cellular aggregation formation. Additionally, the expression of IL-2 and IFN-r were up-regulated after the treatment of rFip-gts, and a corresponding increased production of IL-2 and IFN-r in a dose-dependent manner. The results showed that rFip-gts has an immunomodulatory activity in inducing Th1 lymphocytes from murine splenocytes released IL-2 and IFN-γ, thus suggest that rFip-gts may have therapeutic potential in vivo as an immune modulator.
Abstract: The avian phytohaemagglutinin skin test is being
proved as an in vivo system for the evaluation an avian in vivo T cell
mitogenicity. The test system was one week old Gallus domesticus
broiler Chickens. Five replicates were done for each of the whole,
1:10 dilutions of each of 0.05 IU tuberculin, tetanus immunoglobulin
and DPT vaccine as test materials. The evaluation parameters were
the skin indurations and lymphoblast percentages in bone marrow
lymphocytes.
Tuberculin indurations were 2.06 and 1.26mm for 0.05 IU
respectively while lymphoblast percent were 0.234 and 0.1
accordingly.
The skin indurations of 135mg/ml and 1.35mg/ml tetanus
immunoglobulin were 4.86 and 3.96mm while lymphoblast
percentages were 0.3 and 0.14 respectively.
The whole DPT and 1:10 concentration were with 4.5 and 3.2mm
while their lymphoblast percentages were 0.28 and 0.12 accordingly.
Thus the mitogenicity of the test materials was of dependant type.
Abstract: The aim of this work was to study the in vitro effects
of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative
responses of human lymphocytes and Th1 and Th2 cytokine
secretion. The possible protective role of vitamin E on intracellular
stress oxidative induced by these compounds was also investigated.
Peripheral blood lymphocytes were isolated using differential
centrifugation on a density gradient of Histopaque. They were
cultured with mitogen concanavalin A, vitamin E (10 μM) and with
different concentrations of the compounds 1 and 2 (0.1 to 10 μM).
Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits),
intracellular superoxide anion were determined. 1 and 2 were
immunostimulant and increased cytokine secretion with a shift away
from Th1 response to Th2. These properties were however
accompanied by an increase in intracellular oxidative stress. The
presence of vitamin E exhibited protective effects by reducing δ-
lactam-induced superoxide anion generation in lymphocytes.