Histopathological features of peripheral T-cell lymphoma in Sprague Dawley rats induced with N-methyl-N-nitrosourea

This study described the histopathological features of peripheral T-cell lymphoma in male Sprague Dawley rats following intraperitoneal (i.p.) injections of N-methyl-N-nitrosourea (MNU) at a dose of 60 mg/kg body weight per injection, administered twice weekly for 2 consecutive weeks, and followed by a five-month’s observation period. Control rats were injected with normal saline, i.p. All the rats treated with MNU had enlargement of lymph nodes, with 30% had hepatosplenomegaly and 7% had enlarged kidneys at necropsy. Malignant lymphoma was observed in the lymph nodes, spleen, liver, lung, heart, and kidneys. The neoplastic cells were characterised as undifferentiated, and small to large size with bizarre pleomorphic nuclei. The severity was further described as mild, moderate and severe, based on the diffuseness of the lesions. Nonetheless, similar lesions were not observed in the thymus of the rats. Immunohistochemistry staining of the organs was positive for CD3 antibody, which is consistent with T-cell lymphoma

Detection of Bcl-2 gene in leukaemic rats using an EvaGreen real-time RT-PCT assay

Bcl-2 is an anti-apoptotic gene that is involved in the apoptosis process. Suppression of apoptosis by anti- apoptotic gene can contribute to the occurrence of diseases such as leukaemia. The objectives of this study were 2-folds: first, to compare the sensitivity of an EvaGreen quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) with a conventional RT-PCR for the amplification of the Bcl-2 gene; second, to determine the expression of the Bcl-2 gene in N-methyl-N-nitrosourea (MNU)-induced leukaemiain rats using the EvaGreen qRT-PCR assay. A total of 32 male Sprague Dawley rats were assigned into two groups (n=16), namely, control and MNU groups. In particular, MNU was administered intraperitoneally (i.p) at a dose of 60 mg/kg body weight per injection at two times per week for 2 consecutive weeks. The rats were sacrificed after five months and blood samples were collected for RNA extraction and haemogram. The RNAs were converted into cDNA and amplified using both the EvaGreen qPCR and the conventional PCR assays. All the results were normalised with a housekeeper gene, i.e. glyceraldehyde 3-phosphate dehydrogenase (GADPH). The products of amplification were run on gel electrophoresis and all the results were then compared. Based on the relative intensity of the bands, the EvaGreen qRT-PCR assay was highly sensitive compared to the conventional RT-PCR assay as the Bcl-2 gene could not be amplified using the conventional RT-PCR. Interestingly, the results in this study showed that the expression of Bcl-2 was higher in rats with marked lymphocytosis as compared to the leukaemic rats with normal to mildly increase in lymphocyte count. In conclusion, EvaGreen qRT-PCR assay is more sensitive compared to the conventional RT-PCR, and Bcl-2 gene is abundantly expressed in leukaemic rats with marked lymphocytosis compared to the leukaemic rats with normal to mildly increase in lymphocyte number

Brewers rice, a by-product from rice processing, has antiproliferative activity on human colorectal cancer (HT-29) cell line

Colon carcinogenesis is a malignant tumor, and is well-known as the third leading cancer, which contributes to high mortality and morbidity worldwide. Brewers’ rice, known locally as temukut, consists of a mixture of broken kernels with rice bran and rice germ which is a by-product produced in the rice industry. Although extensive studies on the anti-cancer properties of rice bran, published data on the cytotoxicity of brewers’ rice are very limited. The present study was conducted to evaluate the apoptosis induction capability of the water extract of brewers’ rice (WBR) on human colorectal cancer (HT-29) cell line. The HT-29 cells were treated with various concentrations (16, 32, and 64 μg/mL) of WBR for 24 and 48 hours. The morphological analysis of apoptotic cells was evaluated using inverted light microscope and fluorescence microscope. The apoptotic HT-29 cells was evaluated using Annexin V-FITC and propidium iodide (PI) staining apoptosis test and cell cycle analyses. The data obtained were evaluated using a one-way analysis of variance (ANOVA) and P < 0.05 was considered statistical significant. Overall analyses indicated that WBR induced typical characteristics of apoptosis in HT-29 cells, including nuclear fragmentation (NF), nuclear compaction (NC), apoptotic bodies (AB), cellular shrinkage (CS), and chromatin condensation (CC), as visualized under inverted light microscope and fluorescence microscope. Cell cycle analyses and Annexin V-FITC and propidium iodide (PI) staining apoptosis test using flow cytometry revealed that WBR induced apoptotic population in HT-29 cells. In this study, our findings provide clear evidence that WBR inhibits the growth of HT-29 cells via induction of apoptosis. Taken together, we suggest that WBR may be a potential candidate for the prevention and treatment of colorectal cancer

LOW LEVEL OF CYCLOOXYGENASE-2 TRANSCRIPT IN THE SPLEEN OF LYMPHOMA RATS SUPPLEMENTED WITH GARLIC POWDER

Cyclooxygenases (COXs) regulate tumor cell proliferation and metastasis in many types of cancers including hematological cancers. Organosulfursderived from garlic have a potential to inhibit the expression of COX-2 in cancer patients.Objective: In this study, we evaluate the transcription levels of COX-2 in the spleen of lymphoma rats supplemented with garlic powder.Methods: Three groups of rats were equally divided into control (n=3), lymphoma (n=3), and lymphoma supplemented with garlic powder (n=3)groups. Lymphoma was induced via administration of N-methyl-N-nitrosourea (MNU) intraperitoneally 4 times in 2 week periods. Garlic powdermixed with ground commercial rat diet was given daily at 5% of feed intake, starting at day 1 of MNU exposure. All rats were kept for 24 weeks beforespleen samples were collected and extracted for total RNA. The transcription levels of COX-2 transcript in the total RNA were determined usingquantitative real-time reverse transcriptase polymerase chain reaction assay. The total RNA was converted into cDNA followed by amplification ofCOX-2 and beta-actin genes.Results: Results of the amplification of COX-2 transcripts were normalized with the housekeeping gene, beta-actin. The relative transcription levelof COX-2 transcript in the spleen of lymphoma rats was 1.941±0.131 fold higher (p&lt;0.05) than control rats (1.00±0.001 fold), while the transcriptionlevels in the spleen of lymphoma rats supplemented with garlic was significantly lower (0.423±0.239 SE fold) than the lymphoma rats that receivedno supplementation of garlic powder.Conclusion: The findings suggest that garlic powder reduces the transcription of COX-2 transcript in the spleen of lymphoma ratsKeywords: Garlic, Cyclooxygenase-2, N-methyl-N-nitrosourea, Splenic lymphoma, Real-time reverse transcriptase polymerase chain reaction assay

Haematological parameters of leukaemic rats supplemented with Morinda citrifolia

Morinda citrifolia (mengkudu) had been reported to have anti-tumor activity, which has been researched widely in several animal models. Sixty four eight-week-old male Sprague Dawley rats were divided into four groups of 16 rats per group namely control, MNU, Morinda citrifolia (MC), and MC+MNU group. The MNU and MC+MNU groups received four consecutive intraperitoneal (i.p) injections of N-methyl-N-nitrosourea (MNU) at a dose of 60 mg/kg for induction of leukaemia. Rats in the MC and MC+MNU groups were fed daily with a ration mixed with M. citrifolia at a dose of 5000 mg/kg body weight. The peripheral blood samples were collected at 20 weeks post MNU administration into EDTA tubes and analysed for a complete blood count. Blood smears stained with Wright’s stain were prepared for a manual differential leukocyte count and examination of the leukaemic cells. The results were analysed using a one-way ANOVA. Results in this study showed that MNU group had significant lymphocytosis (66.9±98.14) compared to the other groups. The morphology of the lymphocytes in the MNU and MC+MNU groups showed a typical morphology of leukaemic cell, while the other groups had normal lymphocyte morphology. Rats in the MNU group also had anaemia with significant reduction in total erythrocyte number (6.11±2.73 x 1012/L), haemoglobin concentration (131.64±21.32 g/L) and packed cell volume (37.92±8.50 L/L). The erythron parameters of MC+MNU group were comparable to the control and MC groups. In conclusion, daily supplementation of M. citrifolia reduced the proliferation of circulating leukaemic cells

Physical and nutritional properties of Malaysian avocado (Persea americana Mill) fruit

The tropical climate, along with abundant of rainfall throughout the years, produces a wide diversity of succulent and edible fruits in the Malaysia. In Malaysia, avocado (Persea americana Mill) is an underutilized fruit grown in semi-wild conditions of the rural areas or in the small orchards along with other commercial fruit trees. As the global market for avocado fruit is increasing, the underutilized avocado has the potential to be developed as a new commercial fruit crop in Malaysia. Since data on the physical and nutritional properties of Malaysian avocado fruit are necessary for the food industry to determine which part of the fruit can be used for commercialization, a study was conducted with this aim in mind. Results indicated the Malaysian avocado is a medium sized fruit with an average mass, length and diameter of 216.92 g, 9.50 cm and 7.20 cm, respectively. The major part of Malaysia avocado fruit is the pulp (56.01%), followed by the seed (33.04%) and the peel (10.94%). Each of these parts was further investigated for their nutrient composition. All the fruit parts were found to be low in protein ( 60%). Compared to the peel and seed, the pulp has the highest lipid content (20.79 ± 0.27%), but the lowest ash (1.47 ± 0.13%) and carbohydrate (3.39 ± 0.76%) contents. Malaysian avocado fruit, particularly the pulp, contains a significant amount of lipid content, suggesting the potential for extraction of an edible oil

Antihypercholesterolemic and antioxidant efficacies of zerumbone on the formation, development, and establishment of atherosclerosis in cholesterol-fed rabbits

Hassan Othman Hemn,1,2 Muhammad Mustapha Noordin,1 Heshu Sulaiman Rahman,1,2 Hamza Hazilawati,1 Abubakr Zuki,1 Max Stanley Chartrand3 &nbsp;1Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia; 2College of Veterinary Medicine, University of Sulaimani, Sulaimani City, Kurdistan, Republic of Iraq; 3DigiCare, Behavioral Research, Casa Grande, AZ, USA&nbsp;Abstract: Owing to the high incidence of cholesterol-induced cardiovascular disease, particularly atherosclerosis, the current study was designed to investigate the preventive and therapeutic efficacies of dietary zerumbone (ZER) supplementation on the formation and development of atherosclerosis in rabbits fed with a high cholesterol diet. A total of 72 New Zealand white rabbits were divided randomly on two experimental studies carried out 8 weeks apart. The first experiment was designed to investigate the prophylactic efficacy of ZER in preventing early developed atheromatous lesion. The second experimental trial was aimed at investigating the therapeutic effect of ZER in reducing the atherosclerotic lesion progression and establishment. Sudanophilia, histopathological, and ultrastructural changes showed pronounced reduction in the plaque size in ZER-medicated aortas. On the other hand, dietary supplementation of ZER for almost 10 weeks as a prophylactic measure indicated substantially decreasing lipid profile values, and similarly, plaque size in comparison with high-cholesterol non-supplemented rabbits. Furthermore, the results of oxidative stress and antioxidant biomarker evaluation indicated that ZER is a potent antioxidant in suppressing the generation of free radicals in terms of atherosclerosis prevention and treatment. ZER significantly reduced the value of malondialdehyde and augmented the value of superoxide dismutase. In conclusion, our data indicated that dietary supplementation of ZER at doses of 8, 16, and 20 mg/kg alone as a prophylactic measure, and as a supplementary treatment with simvastatin, significantly reduced early plague formation, development, and establishment via significant reduction in serum lipid profile, together with suppression of oxidative damage, and therefore alleviated atherosclerosis lesions.Keywords: zerumbone, antihypercholesterolemic, antioxidant, atherosclerosis, rabbit mode

Pathological changes and bacteriological assessments in the urinary tract of pregnant goats experimentally infected with Brucella melitensis

Abstract Background This study was conducted to investigate the pathological changes and distribution of B. melitensis in the urinary tract of pregnant goats following acute experimental infection. Six Jamnapari crossbred does in their third trimester of pregnancy were randomly assigned into two groups; Group 1 was uninfected control and Group 2 was inoculated conjunctival with 0.1 mL of the inoculums containing 109 cfu/mL of live B. melitensis. All does were sacrificed 30 days post-inoculation before the kidney, ureter, urinary bladder, urethra and vaginal swab were collected for isolation of B. melitensis. The same tissue samples were fixed in 10% neutral buffered formalin for hematoxylin and eosin, and immunoperoxidase staining. Results None of the goats showed clinical signs or gross lesions. The most consistent histopathology finding was the infiltration of mononuclear cells, chiefly the macrophages with few lymphocytes and occasionally neutrophils in all organs along the urinary tract of the infected goats of Group 2. Other histopathology findings included mild necrosis of the epithelial cells of the renal tubules, congestion and occasional haemorrhages in the various tissues. Kidneys showed the most severe lesions. Immunoperoxidase staining revealed the presence of B. melitensis within the infiltrating macrophages and the epithelium of renal tubules, ureter, urethra and urinary bladder. Most extensive distribution was observed in the urinary bladder. Brucella melitensis was successfully isolated at low concentration (3.4 × 103 cfu/g) in the various organs of the urinary tract and at high concentration (2.4 × 108 cfu/mL) in the vaginal swabs of all infected goats. Although B. melitensis was successfully isolated from the various organs of the urinary tract, it was not isolated from the urine samples that were collected from the urinary bladder at necropsy. Conclusion This study demonstrates the presence of low concentrations of B. melitensis in the organs of urinary tract of pregnant does, resulting in mild histopathology lesions. However, B. melitensis was not isolated from the urine that was collected from the urinary bladder

The induction of aryl hydrocarbon receptor (AHR) in immune organs of developing chicks by polycyclic aromatic hydrocarbons

<p>Polycyclic aromatic hydrocarbons are pollutants which are persistent in nature. The aryl hydrocarbon receptor is a ligand-activated cytosolic transcription factor activated by xenobiotics. The objective was to isolate and identify AHR mRNA transcript in immune organs of developing chicks and to interpret the correlation between AHR induction and dose of PAHs. Specific pathogen free embryonated eggs on day nine were inoculated with solutions of pyrene, phenanthrene, and fluoranthene dissolved in tricaprylin (vehicle) through the allantoic route at three dose levels: 0.2 mg/kg, 2 mg/kg, and 20 mg/kg. A 650 base pair product was observed by RNA extraction and reverse transcription PCR from thymus, bursa of Fabricius and spleen on 21st day. When AHR concentration was analyzed by ELISA in these organs, pyrene showed maximum potency in inducing AHR in thymus. Fluoranthene made highest concentration of AHR in bursa of Fabricius. None of these chemicals caused an increase in AHR concentration in spleen.</p

Chemopreventive effects of garlic powder on N-methyl-N-nitrosourea-induced ductal mammary gland carcinoma in female Sprague Dawley rats

The current study was conducted to investigate and evaluate the chemopreventive effects of garlic powder on MNU induced mammary glands carcinoma in female Sprague Dawley rats. Twenty four, 8-week old female Sprague Dawley rats were assigned randomly into 3 groups each group 8 rats namely, A, B and C. Group A served as a control group. Groups B and C received four consecutive subcutaneous injections of N-methyl-N-nitrosourea (MNU) at a dose of 60 mg/kg of body weight during the first two weeks (2 injections per week) of the experimental period. Group C rats were received diet containing 5% of garlic powder daily during 24 weeks of the experimental period. All rats were weighed and euthanized at the end of the experimental period by bleeding under general anaesthesia. Gross pathology examination was conducted immediately after necropsy and mammary glands were collected for histopathology analysis. All MNU treated rats of groups B and C had shown decrease in body weights in comparison to group A rats. Mammary glands enlargement was observed in 100% of group B rats and 37.5% of group D rats. Ductal mammary gland carcinoma lesion was observed in all enlarged mammary glands of groups B and C rats. In conclusion, administration of four subcutaneous injections of MNU induced ductal mammary gland carcinoma in female Sprague Dawley rats and daily intake of diet containing 5% of garlic powder for 24 weeks reduced MNU-induced ductal mammary gland carcinoma in female Sprague Dawley rats