Displaying publications 1 - 20 of 47 in total

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  1. Yehya AHS, Subramaniam AV, Asif M, Kaur G, Abdul Majid AMS, Oon CE
    World J Gastroenterol, 2022 Aug 28;28(32):4620-4634.
    PMID: 36157930 DOI: 10.3748/wjg.v28.i32.4620
    BACKGROUND: Pancreatic cancer is the most aggressive cancer type. Gemcitabine is the first line chemo-drug used for pancreatic cancer but exerts a broad spectrum of organ toxicities and adverse effects in patients.

    AIM: To evaluate the anti-tumour activity and toxicological effects of Orthosiphon stamineus extract formulation (ID: C5EOSEW5050ESA trademarked as Nuva-staticTM), and gemcitabine combination on pancreatic xenograft model.

    METHODS: Mice were randomly divided into six groups of 6 mice each (n = 6) and given different treatments for 28 d. The study design consisted of a 2 x 3 factorial treatment structure, with gemcitabine (yes/no) by oral (at 1200 and 400 mg/kg per day). Human pancreatic cancer cells were injected subcutaneously into the flanks of athymic nude mice. C5EOSEW5050ESA (200 or 400 mg/kg per day) was administered orally, while gemcitabine (10 mg/kg per 3 d) was given intraperitoneally either alone or in combination treatment. Histopathological analyses of vital organs, tumour tissues, and incidence of lethality were analysed. Analyses of tumour necrosis and proliferation were determined by haematoxylin-eosin staining and immunohistochemistry for Ki-67, respectively.

    RESULTS: No signs of toxicity or damage to vital organs were observed in all treatment groups compared to the untreated group. C5EOSEW5050ESA at 200 mg/kg and gemcitabine combination had no additive antitumor effects compared to a single treatment. Remarkably, a comparably greater response in a reduction in tumour growth, Ki-67 protein expression, and necrosis was demonstrated by 400 mg/kg of C5EOSEW5050ESA and gemcitabine combination than that of the individual agents.

    CONCLUSION: These results highlighted the synergistic activity of C5EOSEW5050ESA with gemcitabine to reduce pancreatic tumour growth in mice compared to a single treatment. Thus, this study provides valuable insights into using C5EOSEW5050ESA as a complementary treatment with gemcitabine for pancreatic cancer.

  2. Tobe SS, Swaran YC, Dennany L, Sibbing U, Schulze Johann K, Welch L, et al.
    Int J Legal Med, 2017 Jan;131(1):87-94.
    PMID: 27832353 DOI: 10.1007/s00414-016-1461-x
    Sexual assault samples are some of the most common samples encountered in forensic analysis. These samples can require a significant time investment due to differential extraction processes. We report on the first record of successful direct amplification of semen for STR analysis. Neat seminal fluid, dilutions ranging from 1:5 to 1:160 and GEDNAP samples were successfully amplified using a direct method. A mild differential isolation technique to enrich spermatozoa was developed and successfully implemented to separate and directly amplify a mixture of semen and female epithelial cells. Aliquots of samples subjected to the differential isolation protocol were stained with Haemotoxylin and Eosin for sperm scoring. Samples stained after PCR showed a complete lack of intact spermatozoa demonstrating that the cells are lysed during the PCR process. This paper demonstrates the potential to incorporate direct PCR in cases of sexual assault to more rapidly obtain results and achieve a higher sensitivity.
  3. Ezlan Elias, Khairul Osman, Sharifa Abdul Aziz, Abdul Halim Mansar, Siti Fatimah Ibrahim, Jamaludin Mohamed
    MyJurnal
    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.
  4. Ambu, Stephen, Yin, Evelyn Synn Yeoh, Joon, Wah Mak, Chakravarthi, Srikumar
    MyJurnal
    The aim of the study was to assess the prevalence of Sarcocystis infection in wild and peri-urban rodents in some states in Peninsular Malaysia. The thigh muscle from these rodents were formalin preserved, sectioned and stained with hematoxylin and eosin and examined under light microscopy. Of the 146 muscle tissue examined only 73 were positive for Sarcocystis infection.
    Morphological identification showed the presence of some new morphological types to be present. Different species of Sarcocystis were seen in the sections but more extensive studies are needed to identify them to species level.
  5. Ramlah Zainudin, Elvy Quatrin Deka, Lela Su’ut, Hasnizam Abdul Wahid
    Sains Malaysiana, 2015;44:1289-1299.
    Studies have shown that some characters, such as morphological, biochemical and behavioural characteristics were shared among con-specifics of closely related frog species. It is hypothesized that closely related frogs would have similar vocal apparatus structure to produce similar call characteristics, in order to be recognized by con-specifics. The vocal apparatus structures of the frogs from Ranidae were, thus, investigated and calls were recorded from various locations in Sarawak, Borneo. The vocal apparatus was sliced and stained using haematoxylin and eosin staining method. The prepared slides were scanned using Dotslide System Olympus BX51. Captured images were further measured using Microscope Olympus BX51. The results showed that the vocal apparatus structure of ranids in Sarawak differed in length between pulvinar vocale and posterior margin (pvpm) or known as vocal cord stricter. This character was deemed to be the most significant character to discriminate the Sarawak ranids. It is suggested that histological analysis is also a good method for study of phenotypic variation of frog species, as shown by the vocal apparatus structure of the anurans. This study provides an alternative method to differentiate taxonomic identity among frog species.
  6. Guo HF, Mohd Ali R, Abd Hamid R, Chang SK, Zainal Z, Khaza'ai H
    Int J Burns Trauma, 2020;10(5):218-224.
    PMID: 33224609
    Burns are injuries on the skin or other tissues. Burns are divided into superficial, partial, and full-thickness, characterized by the depth of the affected tissues. Histological analysis is critical to assess the burn wound healing process. Thus, a systematic evaluation system is imperative for burn research. In the present study, a total of thirty Sprague-Dawley rats were randomly divided into five groups. Deep partial-thickness burn wound was induced on the dorsal part of the rats. Six animals from each group were sacrificed on the 3rd, 7th, 11th, 14th and 21st day post-burn, respectively. Half of the wound tissue was immediately fixed in buffered neutral formalin for hematoxylin & eosin staining. The healing of the epidermis was evaluated with scores ranging from 0 to 7 based on the state of crust on wound surface, the degree of epithelialization as well as the formation of rete ridges. Meanwhile, healing of the dermis was also evaluated with scores ranging from 0 to 7 according to the proportion of adipose cells, inflammatory cells and fibroblasts, the state of collagen deposition as well as the formation of hair follicles. Furthermore, temporal changes of histological score of epidermis and dermis in the skin tissue with deep partial-thickness burn was evaluated. In conclusion, a new comprehensive system for assessing microscopic changes in the healing process of deep partial-thickness burn wound in hematoxylin & eosin staining slides was established, which simplified the scoring process and helped to obtain reproducible and accurate results in the burn study.
  7. Nur Ain, J., Ainaa Diyana, S., Nur Hazirah, R., Nur Fathiah, A.S., Fairus, A., Teoh, S.l., et al.
    Medicine & Health, 2018;13(1):165-174.
    MyJurnal
    Piper betle (PB) leaves have been traditionally used in many Asian countries for the healing of wounds and other ailments. The aim of this study was to examine the wound healing effect of PB on skin injury-inflicted Sprague-Dawley rats. Eighteen male Sprague-Dawley rats were inflicted with wounds and then divided into three groups consisting of a control (normal wound healing without dressing), saline (wound healing aided by gauze soaked in saline) and PB (gauze impregnated with PB paste) dressings. Wounds were created on the back of rats with 6-mm sterilized punch biopsy needle. Wounds were examined on days 3, 5, 7, 9, 11 and 13 for dryness, exudation, and scar formation. Rats were sacrificed on day 14 and the granulation tissue formed on the wound was then excised for histological examination using Hematoxylin and Eosin staining. The formation of granulation tissue in rats treated with PB showed higher progress to wound closure with shorter days (on day 3) compared to the control and saline group (on day 5). PB dressing minimized the formation of scar tissue significantly (p
  8. Agarwal A, Singh S, Agarwal S, Gupta S
    Malays Orthop J, 2018 Nov;12(3):31-37.
    PMID: 30555644 DOI: 10.5704/MOJ.1811.007
    Introduction: Early diagnosis of osteoarticular tuberculosis (OATB) is essential to prevent significant functional disability. There is no single test for diagnosis. Despite an array of investigations available, definitive diagnosis at early stage before starting antitubercular drugs is still a challenge. Materials and Methods: A cross sectional study was carried out between February 2016 and October 2017. All children less than 18 years of age with suspected osteoarticular tuberculosis were included. The cases were subjected to simple needle aspiration from whichever site was accessible. Multiple sample aspirations were done at site of involvement. Smears were prepared from the aspirated material. Results: Ziehl-Neelsen staining for Acid Fast Bacilli (AFB) showed deep pink red rods under light microscopy. Features suggestive of tuberculosis can be seen by May-Grünwald-Giemsa (MGG) staining. Auramine-O staining method of detecting AFB under fluorescent microscope shows the bacilli as greenish yellow slender curved rods in dark background. Fluorescent microscopy has higher sensitivity and comparable specificity. In our study, microbiological confirmation of OATB could be done in 100% cases where the lesion could be accessed for aspiration. The molecular techniques are relatively more expensive and not available everywhere. Conclusion: Meticulous search for AFB in a well stained smear using three different staining methods provides a direct evidence of infection over costly imaging especially in poor patients seen in resource limited settings.
  9. Golkhandan E, Kamaruzaman S, Sariah M, Abidin MAZ, Nazerian E, Yassoralipour A
    Plant Dis, 2013 May;97(5):685.
    PMID: 30722205 DOI: 10.1094/PDIS-08-12-0759-PDN
    In August 2011, sweet potato (Ipomoea batatas), tomato (Solanum lycopersicum), and eggplant (S. melongena) crops from major growing areas of the Cameron highlands and Johor state in Malaysia were affected by a soft rot disease. Disease incidence exceeded 80, 75, and 65% in severely infected fields and greenhouses of sweet potato, tomato, and eggplant, respectively. The disease was characterized by dark and small water-soaked lesions or soft rot symptoms on sweet potato tubers, tomato stems, and eggplant fruits. In addition, extensive discoloration of vascular tissues, stem hollowness, and water-soaked, soft, dark green lesions that turned brown with age were observed on the stem of tomato and eggplant. A survey was performed in these growing areas and 22 isolates of the pathogen were obtained from sweet potato (12 isolates), tomato (6 isolates), and eggplant (4 isolates) on nutrient agar (NA) and eosin methylene blue (EMB) (4). The cultures were incubated at 27°C for 2 days and colonies that were emerald green on EMB or white to gray on NA were selected for further studies. All bacterial cultures isolated from the survey exhibited pectolytic ability on potato slices. These bacterial isolates were gram negative; rod shaped; N-acetylglucosaminyl transferase, gelatin liquefaction, and OPNG positive; and were also positive for acid production from D-galactose, lactosemelibiose, raffinose, citrate, and trehalose. They were negative for indol production, phosphatase activity, reducing substances from sucrose, and negative for acid production from maltose, sorbitol, inositol, inolin, melezitose, α-mathyl-D-glocoside, and D-arabitol. The bacteria did not grow on NA at 37°C. Based on these biochemical and morphological assays, the pathogen was identified as Pectobacterium wasabiae (2). In addition, DNA was extracted and PCR assay with two primers (16SF1 and 16SR1) was performed (4). Partial sequences of 16S rRNA (GenBank Accession Nos. JQ665714, JX494234, and JX513960) of sweet potato, tomato, and eggplant, respectively, exhibited a 99% identity with P. wasabiae strain SR91 (NR_026047 and NR_026047.1). A pathogenicity assay was carried out on sweet potato tubers (cv. Oren), tomato stems (cv. 152177-A), and eggplant fruits (cv. 125066x) with 4 randomly representative isolates obtained from each crop. Sweet potato tubers, tomato stems, and eggplant fruits (4 replications) were sanitized in 70% ethyl alcohol for 30 s, washed and rinsed in sterile distilled water, and needle punctured with a bacterial suspension at a concentration of 108 CFU/ml. Inoculated tubers, stems, and fruits were incubated in a moist chamber at 90 to 100% RH for 72 h at 25°C when lesions were measured. All inoculated tubers, stems, and fruits exhibited soft rot symptoms after 72 h similar to those observed in the fields and greenhouses and the same bacteria were consistently reisolated. Symptoms were not observed on controls. The pathogenicty test was repeated with similar results. P. wasabiae have been previously reported to cause soft rot on Japanese horseradish (3), and aerial stem rot on potato in New Zealand (4), the U.S. (2), and Iran (1). To our knowledge, this is the first report of sweet potato, tomato, and eggplant soft rot caused by P. wasabiae in Malaysia. References: (1) S. Baghaee-Ravari et al. Eur. J. Plant Pathol. 129:413, 2011. (2) S. De Boer and A. Kelman. Page 56 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. N. Schaad et al., eds. APS Press, St. Paul, 2001. (3) M. Goto et al. Int. J. Syst. Bacteriol. 37:130, 1987. (4) A. R. Pitman et al. Eur. J. Plant Pathol. 126:423, 2010.
  10. Sarmadi S, Izadi-Mood N, Sanii S, Motevalli D
    Malays J Pathol, 2019 Apr;41(1):15-24.
    PMID: 31025633
    INTRODUCTION: In the event of encountering hydropic villi in products of conception specimens, pathologists will have to distinguish complete and partial hydatidiform mole (CHM & PHM) from hydropic abortion (HA). The histological diagnostic criteria are subjective and demonstrate considerable inter-observer variability.

    MATERIALS AND METHODS: This study evaluated the inter-observer variability in diagnosis of CHM, PHM and HA according to defined histologic criteria. Ninety abortus conception specimens were reviewed. Representative haematoxylin and eosin-stained slides were assigned independently to two pathologists who were asked to make a diagnosis of CHM, PHM or HA, and provide a report of the identified diagnostic histological criteria. Kappa value was calculated for the inter-observer agreement.

    RESULTS: There was a total of 36.7% disagreement between two pathologists (K = 0.403, Strength of Agreement = moderate), of which 24.4% and 12.2%, were differentiating PHM from CHM and PHM from HA, respectively. Among defined diagnostic histological criteria, the highest rate of agreement was observed in the identification of cistern formation and hydropic changes (K = 0.746 and 0.686 respectively, Strength of Agreement = substantial).

    CONCLUSION: There was moderate to substantial agreement rate between two pathologists in identification of two essential histologic criteria for diagnosis of molar pregnancies i.e. "hydropic change" and "trophoblastic proliferation".

  11. Oktiansyah R, Juliandi B, Widayati KA, Juniantito V
    Trop Life Sci Res, 2018 Jul;29(2):1-11.
    PMID: 30112137 DOI: 10.21315/tlsr2018.29.2.1
    Neuronal cell death can occur in a tissue or organ, including the brain, which affects memory. The objectives of this study were to determine the dose of bee venom that causes neuronal death and analyse the alteration of mouse behaviour, focusing in particular on spatial memory. Fifteen male mice of Deutsche Denken Yoken (DDY) strain were divided into control and treatment groups. Bee venom was injected six times for two weeks intraperitoneally with 1.88 mg/kg, 3.76 mg/kg, 5.6 mg/kg, and 7.48 mg/kg doses of venom. Brain histology was studied using haematoxylin-eosin stained paraffin embedded 5 μm coronal sections. A Y maze test was used to assay behaviour. Parameters observed were the number of dead neurons and the percentage of mice with altered behaviour. ANOVA showed that the effects of bee venom were significantly different in the case of the neuronal death parameter but were not significantly different in the case of the mice behaviour parameter. Duncan's Multiple Range Test (DMRT) demonstrated that P4 (7.48 mg/kg) gave the highest effect of bee venom to promote neuronal death.
  12. Hock, Eng Khoo, Azrina Azlan, Amin Ismail, Al-Sheraji, Sadek Hassan
    MyJurnal
    Defatted dabai peel contains a high amount of anthocyanin. Anthocyanins are known to prevent several
    types of disease, including cardiovascular-related complications. This study aimed to describe the
    effects of different doses of defatted dabai peel extract by histopathological analyses on lesions in the
    liver, kidney, heart and aorta. Histopathology methods were applied to determine the protective effects
    of defatted dabai peel extracts against hypercholesterolemia-induced oxidative damages to animal
    organs. Haematoxylin and eosin staining was applied for histopathology examination for liver, kidney,
    heart and aorta. Data showed that a high dose of defatted dabai extract (3000 mg per day) applied to
    hypercholesterolemic rabbits for eight weeks had mild protective effect, especially reducing the severity
    of hepatic fibrosis and steatosis of the renal medulla. The high dose of extract supplementation also
    reduced inflammation of aorta and formation of atherosclerosis plaque in the cell wall of right ventricle
    of the heart. The high dose of defatted dabai peel extract could be a protective agent against oxidative
    stress.
  13. Tarig AA, Wahid H, Rosnina Y, Yimer N, Goh YM, Baiee FH, et al.
    Vet World, 2017 Jun;10(6):672-678.
    PMID: 28717321 DOI: 10.14202/vetworld.2017.672-678
    AIM: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL) and virgin coconut oil (VCO) in Tris-based extender on chilled and frozen-thawed bull semen quality parameters.

    MATERIALS AND METHODS: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test.

    RESULTS: The results showed that all the quality parameters assessed were significantly (p<0.05) improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+) in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups.

    CONCLUSION: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.

  14. Maryam Mashmoul, Azrina Azlan, Norhafizah Mohtarrudin, Barakatun Nisak Mohd Yusof, Huzwah Khaza’ai, Mehdi Farzadnia, et al.
    MyJurnal
    Thirty-six adult male SD rats, weighing 200-250 g were used in this study. Dietary obesity was induced by feeding high-fat diet (HFD) for 12 weeks. One group of animal (n=6) served as normal control which received normal diet from the beginning to the end of the study. The other 30 male obese animals induced by HFD were randomly divided into five experimental groups: The group which received high-fat diet considered as a negative control and the other four groups were treated with saffron extract and crocin in low and high dosages of 40 and 80 mg/kg for 8 weeks. At the end of treatment period, kidney and pancreas were removed and stained with hematoxylin and eosin (H&E). There were significant histopathological changes, such as tubular degenerative, vascular congestion and interstitial inflammation in the kidney and extensive pancreatic adipose tissue infiltration in high-fat diet rats group when compared with normal control. The results showed less significant histopathological changes of the kidney and also moderate pancreatic adipose tissue infiltration in the groups treated with crocin. Interestingly saffron extract dose-independently displayed significant renal protective effect while a significant decrease in pancreatic adipose tissue infiltration was observed in obese rat treated with high dose of saffron extract (80 mg/kg). From these results, it can be concluded that consumption of saffron extract reduced the untoward effects of high-fat diet in kidney and pancreas of high-fat diet induced obese rats.
  15. Md Ismail, N.H., Nik Mohd Alwi N.A.
    JUMMEC, 2019;22(1):13-19.
    MyJurnal
    Oral ulcer is a lesion with multifactorial causes and occurs worldwide. The lesion usually resolved within 14
    days, but the pain may have an impact on the quality of patient’s life. Therefore, having a natural derived
    remedy that can reduce healing time would be a great advantage. This study aims to investigate histological
    sections of buccal ulcer on rats treated with aqueous extract of Piper sarmentosum (AEPS). Glacial acetic acid
    was used to induce buccal ulcer on male Sprague Dawley rats. Control group received normal saline while
    the experimental group received AEPS for treatment. On certain days of post-ulcer induction, buccal ulcer
    tissue samples were harvested, sectioned and stained with Hematoxylin and Eosin (H&E). Histological slides
    were examined for inflammation and scored. The inflammation severity reduced from day 2 to day 12. In the
    experimental group there was a statistically significant differences of inflammation score, particularly on day
    2 with a score of (2.8 + 0.2). Neutrophils were less in the experimental group and the tissue debris clearance
    was faster compared to control group. Full reepithelization was observed on both treated tissue sections on
    day 12 with less severe inflammation. Topical application of AEPS is proven to have anti-inflammatory effect
    by reducing the number of neutrophils during inflammation phase of oral ulcer healing.
  16. Aamir K, Khan HU, Hossain CF, Afrin MR, Shaik I, Salleh N, et al.
    PeerJ, 2019;7:e8045.
    PMID: 31772835 DOI: 10.7717/peerj.8045
    Background: Arjunolic acid (AA) is a potent phytochemical with wider pharmacological activities. Despite potential medicinal properties on various in vitro and in vivo studies, there is still a dearth of scientific data related to its safety profile and toxicological parameters. The current study aimed to investigate acute toxicity of AA in normal female Sprague Dawley rats.

    Methods: In this study, AA was administered orally at an individual dose of 300 and 2000 mg/kg body weight to group 1 and 2 respectively, while group 3 served as normal control. All the animals were observed for 2 weeks to determine any behavioral and physical changes. On day 15, blood was collected for hematological and biochemical investigation, later animals from all the three groups were euthanized to harvest and store essential organs for histopathological analysis. Four different staining techniques; hematoxylin and eosin, Masson trichrome, Periodic acid Schiff and Oil O Red were used to investigate any alterations in different tissues through microscopical observation.

    Results: The results of the study showed no morbidity and mortality at two different dosage of AA treatment. Daily food & water intake, body weight, relative organ weight, hematological and biochemical parameters were detected to be normal with no severe alteration seen through microscopical investigation in the structure of harvested tissues. Our findings support the safety profile of AA, which was well tolerated at higher dose. Thus, an in-detail study on the subacute disease model is warranted.

  17. Muhammad-Azam F, Nur-Fazila SH, Ain-Fatin R, Mustapha Noordin M, Yimer N
    Vet World, 2019 Nov;12(11):1682-1688.
    PMID: 32009746 DOI: 10.14202/vetworld.2019.1682-1688
    Background and Aim: Laboratory mice are widely used as a research model to provide insights into toxicological studies of various xenobiotic. Acetaminophen (APAP) is an antipyretic and analgesic drug that is commonly known as paracetamol, an ideal hepatotoxicant to exhibit centrilobular necrosis in laboratory mice to resemble humans. However, assessment of histopathological changes between mouse strains is important to decide the optimal mouse model used in APAP toxicity study. Therefore, we aim to assess the histomorphological features of APAP-induced liver injury (AILI) in BALB/C and Institute of Cancer Research (ICR) mice.

    Materials and Methods: Twenty-five ICR mice and 20 BALB/C mice were used where five animals as control and the rest were randomly divided into four time points at 5, 10, 24 and 48 hours post-dosing (hpd). They were induced with 500 mg/kg APAP intraperitoneally. Liver sections were processed for hematoxylin-eosin staining and histopathological changes were scored based on grading methods.

    Results: Intense centrilobular damage was observed as early as 5 hpd in BALB/C as compared to ICR mice, which was observed at 10 hpd. The difference of liver injury between ICR and BALB/C mice is due to dissimilarity in the genetic line-up that related to different elimination pathways of APAP toxicity. However, at 24 hpd, the damage was markedly subsided and liver regeneration had taken place for both ICR and BALB/C groups with evidence of mitotic figures. This study showed that normal liver architecture was restored after the clearance of toxic insult.

    Conclusion: AILI was exhibited earlier in BALB/C than ICR mice but both underwent liver recovery at later time points.

  18. Zhang Y, Lee SH, Wang C, Gao Y, Li J, Xu W
    Jpn J Clin Oncol, 2020 Jun 24.
    PMID: 32579167 DOI: 10.1093/jjco/hyaa089
    BACKGROUND: Patient-derived xenograft model is a powerful and promising tool for drug discovery and cancer biology studies. The application of previous metastatic colorectal cancer models has been greatly limited by its low success rate and long time to develop metastasis. Therefore, in this study, we aim to describe an optimized protocol for faster establishment of colorectal cancer metastatic patient-derived xenograft mouse models.

    METHODS: Smaller micro tissues (˂150 μm in diameter) mixed with Matrigel were engrafted subcutaneously into NSG mice to generate the passage 1 (P1) patient-derived xenograft. The micro tumours from P1 patient-derived xenograft were then excised and orthotopically xenografted into another batch of NSG mice to generate a metastatic colorectal cancer patient-derived xenograft, P2. Haematoxylin and eosin and immunohistochemistry staining were performed to compare the characters between patient-derived xenograft tumours and primary tumours.

    RESULTS: About 16 out of 18 P1 xenograft models successfully grew a tumour for 50.8 ± 5.1 days (success rate 89.9%). Six out of eight P1 xenograft models originating from metastatic patients successfully grew tumours in the colon and metastasized to liver or lung in the NSG recipients for 60.9 ± 4.5 days (success rate 75%). Histological examination of both P1 and P2 xenografts closely resembled the histological architecture of the original patients' tumours. Immunohistochemical analysis revealed similar biomarker expression levels, including CDH17, Ki-67, active β-catenin, Ki-67 and α smooth muscle actin when compared with the original patients' tumours. The stromal components that support the growth of patient-derived xenograft tumours were of murine origin.

    CONCLUSIONS: Metastatic patient-derived xenograft mouse model could be established with shorter time and higher success rate. Although the patient-derived xenograft tumours were supported by the stromal cells of murine origin, they retained the dominant characters of the original patient tumours.

  19. Puebla-Osorio N, Sarchio SNE, Ullrich SE, Byrne SN
    Methods Mol Biol, 2017;1627:213-222.
    PMID: 28836204 DOI: 10.1007/978-1-4939-7113-8_14
    Mast cells are part of the immune system and characteristically contain histamine- and heparin-rich basophilic granules. While these cells are usually associated with allergy and anaphylaxis, they also promote wound healing and angiogenesis and confer protection against pathogens. The presence of these cells is sometimes indicative of a poor prognosis, especially in skin cancer, pancreatic cancer, and lymphoma. Toluidine blue staining of acid-fast granules is an established method for the identification and quantification of mast cells. Generating detailed information on the location of mast cells within tissues is problematic using this technique and often requires serial sections from adjacent tissue to be separately stained with hematoxylin and eosin (H&E). Staining serial sections is not always possible, particularly if the sample is very small or rare. In such cases, a method of simultaneously identifying and localizing mast cells in a tissue would be advantageous. Toluidine blue and H&E are not commonly combined because H&E includes repetitive washes in water, which may affect the efficacy of the aqueous-soluble toluidine blue. We have developed and tested a novel staining technique that integrates toluidine blue between hematoxylin and eosin in one simple procedure. This protocol works on both frozen and formalin-fixed, paraffin-embedded tissue and readily allows for the identification of purple-stained mast cells against a clean H&E background. This facilitates a more accurate localization and proper counting of mast cells in normal and affected tissue.
  20. D'Souza UJ
    Malays J Med Sci, 2003 Jan;10(1):43-5.
    PMID: 23365499 MyJurnal
    The antimetabolite, 5-fluorouracil is widely used in the treatment of cancers. Although its toxic effects on testis causing germinal epithelial sloughing, tubular atrophy and generation of multinucleated cells were reported, its effect on spermatogenesis has not been studied. Hence the present study was conducted to evaluate the effects of 5-fluorouracil on epididymal sperm count. Male Wistar rats were employed in the study (n=5 per group). The animals were injected (i.p) with five consecutive doses of 5-fluorouracil (10, 20, 30mg/kg b.w) at an interval of 24h and the control with 0.1ml-distilled water. Samples were obtained at 14, 35, 42 and 70 days after injection. Rats were sacrificed, a laparatomy was performed and epididymes were collected in 1ml phosphate buffered saline (pH 7.2), minced, filtered and stained with 1% aqueous eosin Y. An aliquot was taken in leucocyte pipette, diluted with phosphate buffered saline and sperm count was done as per the standard procedure. Data were analyzed by Mann Whitney U test. The results of this study revealed that 5 - fluorouracil significantly decreased the sperm count in a dose- and time-dependent manner.
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