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A comparative histopathological study of systemic candidiasis in association with experimentally induced breast cancer

Choo ZW, Chakravarthi S, Wong SF, Nagaraja HS, Thanikachalam PM, Mak JW, et al.

Oncol Lett, 2010 Jan;1(1):215-222.
PMID: 22966285

Systemic candidiasis is a fungal infection which coupled with solid malignancies places patients at high risk of succumbing to the disease. Few studies have shown evidence of the relationship between systemic candidiasis and malignancy-induced immunosuppression disease especially in breast cancer. At present, animal studies that exclusively demonstrate this relationship have yet to be conducted. The exact causative mechanism of systemic candidiasis is currently under much speculation. This study therefore aimed to demonstrate this relationship by observing the histopathological changes of organs harvested from female Balb/c mice which were experimentally induced with breast cancer and inoculated with systemic candidiasis. The mice were randomly assigned to five different groups (n=12). The first group (group 1) was injected with phosphate buffer solution, the second (group 2) with systemic candidiasis, the third (group 3) with breast cancer and the final two groups (groups 4 and 5) had both candidiasis and breast cancer at two different doses of candidiasis, respectively. Inoculation of mice with systemic candidiasis was performed by an intravenous injection of Candida albicans via the tail vein following successful culture methods. Induction of mice with breast cancer occurred via injection of 4T1 cancer cells at the right axillary mammary fatpad after effective culture methods. The prepared slides with organ tissues were stained with hematoxylin and eosin, periodic acidic schiff and gomori methenamine silver stains for a histopathological analysis. Grading of primary tumour and identification of metastatic deposits, as well as scoring of inflammation and congestion in all the respective organs was conducted. Statistical tests performed to compare groups 2 and 4 showed that group 4 exhibited a highly statistically significant increase in organ inflammation and congestion (p<0.01). The median severity of candidiasis in the kidneys and liver also increased in group 4 as compared to group 2. In conclusion, based on the above evidence, systemic candidiasis significantly increased in mice with breast cancer.
A new histological score grade for deep partial-thickness burn wound healing process

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.
A proof of principal study on the use of direct PCR of semen and spermatozoa and development of a differential isolation protocol for use in cases of alleged sexual assault

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.
Ameliorative Effect of Curcumin on Olanzapine-induced Obesity in Sprague-Dawley Rats

Parasuraman S, Zhen KM, Banik U, Christapher PV

Pharmacognosy Res, 2017 Jul-Sep;9(3):247-252.
PMID: 28827965 DOI: 10.4103/pr.pr_8_17

OBJECTIVE: To evaluate the effect of curcumin on olanzapine-induced obesity in rats.

MATERIALS AND METHODS: Sprague-Dawley (SD) rats were used for experiments. The animals were divided into six groups, namely, normal control, olanzapine control, betahistine (10 mg/kg), and curcumin 50, 100, and 200 mg/kg treated groups. Except the normal control group, all other animals were administered with olanzapine 4 mg/kg intraperitoneally to induce obesity. The drugs were administered once daily, per oral for 28 days. During the experiment, body weight changes and behavior alterations were monitored at regular intervals. At the end of the experiment, blood sample was collected from all the experimental animals for biochemical analysis. Part of the liver and kidney tissues was harvested from the sacrificed animals and preserved in neutral formalin for histopathological studies.

RESULTS: Curcumin showed a significant reduction in olanzapine-induced body weight gain on the rats and improved the locomotor effects. The effect of curcumin on olanzapine-induced body weight gain is not comparable with that of betahistine.

CONCLUSION: This study has shown metabolic alteration effect of curcumin on olanzapine, an antipsychotic drug, treated SD rats.

SUMMARY: Olanzapine is an atypical antipsychotic drug used for the treatment of schizophrenia and bipolar disorder. Obesity is an adverse effect of olanzapine, and the present study was made an attempt to study the effect of curcumin on olanzapine-induced obesity in rats. In this present study, curcumin significantly reduced olanzapine-induced body weight gain in rats. Abbreviations Used: 5HT: 5-hydroxytryptamine, ALP: Alkaline phosphatase, ALT: Alanine transaminase, ANOVA: Analysis of variance, AST: Aspartate transaminase, CMC: Carboxymethyl cellulose, D: Dopamine, H and E: Hematoxylin and Eosin stain, H: Histamine, HDL-C: Highdensity lipoprotein cholesterol, IP: Intraperitoneal, MAO: Monoamine oxidase, NaOH: Sodium hydroxide, SD rats: Sprague Dawley rats, TCs: Total cholesterols, TG: Triglyceride.
Fulltext Anti-tumour activity and toxicological studies of combination treatment of Orthosiphon stamineus and gemcitabine on pancreatic xenograft model

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.
Fulltext Assessing the Immunomodulatory Activity of Ethanol Extract of Sambucus javanica Berries and Leaves in Chloramphenicol-Induced Aplastic Anemia Mouse Model

Putra WE, Rifa'i M

Trop Life Sci Res, 2020 Jul;31(2):175-185.
PMID: 32922674 DOI: 10.21315/tlsr2020.31.2.9

Aplastic anemia, life-threatened disease, is a hematologic disorder characterised by bone marrow hypoplasia. Multiple modalities such as bone marrow transplantation and immunosuppression treatment have been proposed to ameliorate this entity, however it remains ineffective. Sambucus, a group of herb plants, possesses a broad spectrum of medicinal properties such as antioxidant, insulin-like activity, anticancer and antiviral. However, the study about its activity toward aplastic anemia incidence is based on limited data. Thus, the research aim of this study was to evaluate the immunomodulatory activities of Sambucus javanica in chloramphenicol-induced anemia aplastic mouse model. In this present study, BALB/c mice were administrated with chloramphenicol (CMP) to induce aplastic anemia then followed by S. javanica extracts treatment. Additionally, cellular and molecular aspects were evaluated by flow cytometry and Hematoxylin-Eosin staining. Further analysis showed that S. javanica extracts could promote the population number of regulatory T-cells and naive cytotoxic T-cells. Moreover, those extract also reduced the inflammation and necrotic incidence in CMP-induced mouse aplastic anemia model. Together, these results suggest that S. javanica has therapeutically effect to aplastic anemia by altering the immune system as an immunomodulatory agent.
Fulltext Candida sp. as a starter culture for cocoa (Theobroma cacao L.) beans fermentation

Mahazar, N. H., Sufian, N. F., Meor Hussin, A. S., Norhayati, H., Mathawan, M., Rukayadi, Y.

International Food Research Journal, 2015;22(5):1783-1787.
MyJurnal

Two cocoa bean fermentation methods (spontaneous fermentation and the use of starter culture) for 7 days fermentation were compared in terms of safety and quality fermented beans. Candida sp. was used as a starter culture in this study. The safety of the fermented cocoa beans were measured by the growth colonies of pathogenic microorganisms namely Bacillus cereus, Escherichia coli, Salmonella sp., Staphylococcus aureus, and Pseudomonas sp., on Bacillus cereus agar, eosin-methylene blue (EMB) agar, xylose lysine deoxycholate (XLD) agar, Baird-Parker agar (BPA), and Pseudomonas agar, respectively. B. cereus, E. coli and Salmonella sp. were early present in both fermentations. Candida sp.-fermentation showed detection of B. cereus at 5.34 log10 CFU/g and absence after 24 hours of fermentation while in spontaneous-fermentation B. cereus was too few to count. Moreover, the log10 E. coli number in Candida sp.-fermentation and spontaneous-fermentation were reduced from 5.72 to 3.66 and from 7.15 to 4.46 on day 1 to day 3, respectively. There were no presences of pathogenic microorganisms on day 5 and day 7 for both fermentations. In term of quality, proximate analysis of spontaneous-fermentation resulted that the content of moisture, ash, fat, crude protein, crude fibre and carbohydrate was 56.47%, 2.32%, 3.17%, 7.02%, 28.14% and 2.88%, meanwhile for the Candida sp.-fermentation was 53.96%, 2.19%, 3.44%, 8.25%, 25.46% and 6.70%, respectively. This study showed that both fermentations are considered to be safe and there is no significant difference in proximate value in fermented cocoa beans from spontaneous-fermentation and Candida sp.-fermentation.
Fulltext Changes in rats' breast tumor ultrastructure and immune and messenger RNA responses caused by dietary Seaweed (Kappaphycus alvarezii) extract

Bakar NA, Tengku Ibrahim TA, Mohamad Shalan NA, Mohamed S

J Microsc Ultrastruct, 2016 08 21;5(2):70-81.
PMID: 30023239 DOI: 10.1016/j.jmau.2016.08.001

The edible red seaweed Kappaphycus alvarezii or Eucheuma cottonii is commercially cultivated in the pristine tropical seas for carrageenan production. The systemic, cellular, and molecular effects of E. cottonii 50% alcohol extract [seaweed E. cottonii ethanol extract (SECE)] on breast cancer were investigated in a rat model. Mammary tumor was induced by subcutaneously injecting LA7 cells in female rat mammary pads. After 2 weeks of cancer growth, the rats received oral administration of either SECE [150 mg/kg body weight (BW) and 300 mg/kg BW] or tamoxifen. Electron microscopy imaging results confirmed macrophage activity and hematoxylin and eosin staining indicated that tumor histopathological alterations were restored toward normal structures by the seaweed extract. The extract suppressed tumor development and modulated the immune responses. This was evidenced by the microscopic observations, the increased spleen weight, size, spleen CD19 B cells, and blood immunoglobulin G (IgG) levels. The extract also increased the circulating total white blood cells, lymphocytes, segmented neutrophils count, T cells (CD3), T-helper cells (CD4), cytotoxic T cell (CD8), and nuclear factor-kappa beta expressions. The extract enhanced cancer cell death, by upregulating the Birc5, Chk1, and p53 levels and downregulating the tumor growth cellular Mdm2 (transformed mouse 3T3 cell double minute 2) messenger RNA (mRNA) expression. The extract showed no toxicity at 150 mg/kg BW in rats. The lectin-rich SECE showed tumor suppression by enhancing immune responses and upregulating the cancer cell apoptosis mRNA expressions.
Fulltext Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus hubbacki): A preliminary study

Iswadi MI, Ann ZF, Hafiz MM, Hafiz MD, Fahrul FJ, Hajarian H, et al.

Open Vet J, 2012;2(1):109-14.
PMID: 26623302

The Malayan gaur (Bos gaurus hubbacki) or Seladang is classified as vulnerable by the International Union for Conservation of Nature and Natural Resources (IUCN). The Malayan gaur is mainly distributed in the tropical woodlands of Peninsular Malaysia and Southern Thailand. The aim of this study was to collect, analyze and cryopreserve the semen of wild Malayan gaur. Transrectal massage (TM) and electroejaculation (EEJ) technique was applied in semen collection of the Malayan gaur. The semen was then cryopreserved in liquid nitrogen using slow freezing technique. Makler counting chamber was used to evaluate sperm concentration and motility, while the sperm viability and morphology of fresh and post-thaw sperm was determined using eosin-nigrosin staining protocol. As a result, we have successfully collected the Malayan gaur semen using EEJ technique. Sperm motility, viability and morphological changes of the post-thaw semen of Malayan gaur were found undesirable due to the complication of the cryopreservation process. On the basis of current study it can be concluded that Malayan gaur bulls semen can be obtain by EEJ with no evidence of rectal trauma. Optimization of the process of cryopreservation for Malayan gaur sperm is needed to maintain the cryoviability of the good sperm quality. The data generated in this study would be useful in conservation of genetic diversity program for Malayan gaur.
Comparative analysis between multilevel sectioning with conventional haematoxylin and eosin staining and immunohistochemistry for detecting nodal micrometastases with stage I and II colorectal cancers

Wong YP, Shah SA, Shaari N, Mohamad Esa MS, Sagap I, Isa NM

Asian Pac J Cancer Prev, 2014;15(4):1725-30.
PMID: 24641399

Management of patients with stage II colorectal carcinomas remains challenging as 20 - 30% of them will develop recurrence. It is postulated that these patients may harbour nodal micrometastases which are imperceptible by routine histopathological evaluation. The aims of our study were to evaluate (1) the feasibility of multilevel sectioning method utilizing haematoxylin and eosin stain and immunohistochemistry technique with cytokeratin AE1/AE3, in detecting micrometastases in histologically-negative lymph nodes, and (2) correlation between nodal micrometastases with clinicopathological parameters. Sixty two stage I and II cases with a total of 635 lymph nodes were reviewed. Five-level haematoxylin and eosin staining and one-level cytokeratin AE1/AE3 immunostaining were performed on all lymph nodes retrieved. The findings were correlated with clinicopathological parameters. Two (3.2%) lymph nodes in two patients (one in each) were found to harbour micrometastases detected by both methods. With cytokeratin AE1/AE3, we successfully identified four (6.5%) patients with isolated tumour cells, but none through the multilevel sectioning method. Nodal micrometastases detected by both multilevel sectioning and immunohistochemistry methods were not associated with larger tumour size, higher depth of invasion, poorer tumour grade, disease recurrence or distant metastasis. We conclude that there is no difference between the two methods in detecting nodal micrometastases. Therefore it is opined that multilevel sectioning is a feasible and yet inexpensive method that may be incorporated into routine practice to detect nodal micrometastases in centres with limited resources.
Fulltext Dentinogenic Ghost Cell Tumor in a Sumatran Rhinoceros

Salleh A, Zainuddin ZZ, Tarmizi RMM, Yap CK, Jeng CR, Zamri-Saad M

Animals (Basel), 2021 Apr 20;11(4).
PMID: 33923894 DOI: 10.3390/ani11041173

An adult female Sumatran rhinoceros was observed with a swelling in the left infraorbital region in March 2017. The swelling rapidly grew into a mass. A radiograph revealed a cystic radiolucent area in the left maxilla. In June 2017, the rhinoceros was euthanized. At necropsy, the infraorbital mass measured 21 cm × 30 cm. Samples of the infraorbital mass, left parotid gland, and left masseter muscle were collected for histopathology (Hematoxylin & Eosin, Von Kossa, Masson's trichrome, cytokeratin AE1/AE3, EMA, p53, and S-100). Numerous neoplastic epithelial cells showing pleomorphism and infiltration were observed. Islands of dentinoid material containing ghost cells and keratin pearls were observed with the aid of the two special histochemistry stains. Mitotic figures were rarely observed. All the neoplastic odontogenic cells and keratin pearls showed an intense positive stain for cytokeratin AE1/AE3, while some keratin pearls showed mild positive stains for S-100. All samples were negative for p53 and S-100 immunodetection. The mass was diagnosed as a dentinogenic ghost cell tumor.
Detection of Infiltrating Mast Cells Using a Modified Toluidine Blue Staining

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.
Fulltext Determination of time of death based on basic histological stain and immunostain changes

Ezlan Elias, Khairul Osman, Sharifa Abdul Aziz, Abdul Halim Mansar, Siti Fatimah Ibrahim, Jamaludin Mohamed

Jurnal Sains Kesihatan Malaysia, 2004;2(2):63-70.
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.
Fulltext Development of a human salivary gland organoid model

Zulaiha A. Rahman, Colin D. Bingle, Lynne Bingle

Malaysian Journal of Medicine and Health Sciences, 2019;15(107):1-1.
MyJurnal

Introduction: Currently, organoid technology provides a useful tool for modelling human organ development and pathologies in vitro. Salivary gland (SG) organoids developed from mice SG cells display self-organizing properties closely mimic the native organ. Thus, this study would like to investigate the potential of this organoid system to de-velop a human salivary gland in vitro. Methods: Organoids were developed from biopsy samples of normal human sublingual gland tissue. Cells were isolated and cultured in Matrigel at an Air Liquid Interface (ALI) for up to 14 days in an enriched media supplementing with Wnt-3A, R-spondin1, EGF, and FGF2. Specific differentiation factors like TGFβ, BMP, and LIMK inhibitors were added to enriched media for further differentiation studies. Haematoxylin and eosin-stained sections of the cultures were used to visualise growth. RT-PCR, immunohistochemistry and immunoflu-orescence were used to determine the differential expression of cell-specific markers. Results: Human SG organoids developed when the cells were grown in Matrigel at ALI in a defined culture system. The addition of TGFβ inhibitor and all the inhibitors (TGFβ, BMP and LIMK) to the culture media affected SG organoids development by displaying distinct characteristics that closely resemble native glands and expressed specific cell-type markers; BPIFA2, AQP5, CK5 and E-cadherin. The inhibition of BMP signalling demonstrated SG organoids growth more into ductal-like struc-tures and expressed ductal cell marker, CK7. While LIM kinase inhibition signalling showed significantly higher of amylase activity assay. Conclusion: This study certainly offers valuable insight into determining the optimal culture conditions for developing human SG organoids.
Fulltext Effect of Eurycoma longifolia Stem Extract on Uric Acid Excretion in Hyperuricemia Mice

Bao R, Liu M, Wang D, Wen S, Yu H, Zhong Y, et al.

Front Pharmacol, 2019;10:1464.
PMID: 31920654 DOI: 10.3389/fphar.2019.01464

Background:Eurycoma longifolia is a tropical medicinal plant belonging to Simaroubaceae distributed in South East Asia. The stems are traditionally used for the treatment of sexual insufficiency, fever, hypertension, and malaria. Furthermore, it has antidiabetic and anticancer activities. Recently, it has been reported to reduce uric acid, but the mechanism is unclear. Hypothesis/Purpose: The aim of this study is to explore the effect and mechanism of E. longifolia stem 70% ethanol extract (EL) and its active compounds on uric acid excretion. Study Design and Methods: Potassium oxonate (PO) induced hyperuricemia rats model and adenine-PO induced hyperuricemia mice model were used to evaluate the effects of EL. Ultraperformance liquid chromatography was used to determine the levels of plasma or serum uric acid and creatinine. Hematoxylin-eosin staining was applied to observe kidney pathological changes, and western blot was applied to detect protein expression levels of uric acid transporters. Effects of constituents on urate uptake were tested in hURAT1-expressing HEK293T cells. Results: EL significantly reduced serum and plasma uric acid levels at dosages of 100, 200, and 400 mg/kg in hyperuricemia rats and mice, increased the clearance rate of uric acid and creatinine, and improved the renal pathological injury. The protein expression levels of urate reabsorption transporter 1 (URAT1) and glucose transporter 9 were down-regulated, while sodium-dependent phosphate transporter 1 and ATP-binding cassette transporter G2 were up-regulated in the kidney after EL treatment. The quassinoids isolated from EL showed inhibitory effects on urate uptake in hURAT1-expressing HEK293T cells, and the effect of eurycomanol was further confirmed in vivo. Conclusion: Our findings revealed that EL significantly reduced blood uric acid levels, prevented pathological changes of kidney in PO induced hyperuricemia animal model, and improved renal urate transports. We partly clarified the mechanism was related to suppressing effect of URAT1 by quassinoid in EL. This study is the first to demonstrate that EL plays a role in hyperuricemia by promoting renal uric acid excretion.
Fulltext Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen

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.
Fulltext Efficacy of choline and DHA supplements or enriched environment exposure during early adult obesity in mitigating its adverse impact through aging in rats

Prabhu GS, Prasad K, K G MR, Rai KS

Saudi J Biol Sci, 2021 Apr;28(4):2396-2407.
PMID: 33911955 DOI: 10.1016/j.sjbs.2021.01.037

Introduction: The aim of this study was to assess the efficacy of choline and DHA or exposure to environmental enrichment in obese adult and aging rats on alterations in body mass index, serum lipid profile and arterial wall changes, despite stopping high fat diet consumption and interventions during adulthood.
Methods: 21 day old male Sprague Dawley rats were assigned as Experiment-1 & 2 - PND rats were divided into 4 groups with interventions for 7 months (n = 8/group). NC- Normal control fed normal chow diet; OB- Obese group, fed high fat diet; OB + CHO + DHA- fed high fat diet and oral supplementation of choline, DHA. OB + EE- fed high fat diet along with exposure to enriched environment .Experiment-2 had similar groups and interventions as experiment 1 but for next 5 months were fed normal chow diet without any interventions. Body mass index was assessed and blood was analyzed for serum lipid profile. Common Carotid Artery (CCA) was processed for Haematoxylin and eosin, Verhoff Vangeison stains. Images of tissue sections were analyzed and quantified using image J and tissue quant software.
Results: In experiment.1, mean body mass index (p
Establishing metastatic patient-derived xenograft model for colorectal cancer

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.
Evidence of plasmotomy in Blastocystis hominis

Tan TC, Suresh KG

Parasitol Res, 2007 Nov;101(6):1521-5.
PMID: 17701428

Blastocystis hominis has been regarded as an enigmatic parasite as many aspects of its basic biology remain uncertain. Many reproductive processes have been suggested for the organism; however, to date, only the binary fission has been proven. Plasmotomy is one of the modes of reproduction previously suggested to be seen in in vitro cultures. The present study provides trichrome and acridine orange staining evidence for the existence of nucleic acid suggestive of division of nucleus into multinucleate forms with the respective cytoplasm dividing giving rise to two or three progeny B. hominis. Transmission electron micrographs further confirmed that these daughter cells had respective surrounding surface coat, mitochondria, and vacuoles.
First Report of Soft Rot Caused by Pectobacterium carotovorum subsp. carotovorum on Pepper Fruits (Capsicum annuum) in Malaysia

Golkhandan E, Kamaruzaman S, Sariah M, Abidin MZZ, Nasehi A, Nazerian E

Plant Dis, 2013 Aug;97(8):1109.
PMID: 30722490 DOI: 10.1094/PDIS-01-13-0042-PDN

Symptoms of water-soaked lesions and soft rot were first observed in June 2011 on bell pepper fruits (Capsicum annuum cv. Annuum) in the two main regions of pepper production in Malaysia (Cameron Highlands and Johor State). Economic losses exceeded 40% in severely infected fields and greenhouses with the estimated disease incidence of 70%. In pepper fruits damaged by insects, sunscald, or other factors, symptoms initially appeared in the peduncle and calyx tissues and entire fruits were turned into watery masses within 2 to 6 days. Fruits infected in the field tended to collapse and hang on the plant. When the contents leaked out, the outer skin of the fruit dried and remained attached to the plant. Field-grown transplants and infected soil were identified as probable sources of inocula. A total of 50 attached fruits were collected from 10 pepper fields and greenhouses located in the two growing regions. Tissue from the margins of water-soaked lesions was surface-sterilized in 1% NaOCl for 2 min, rinsed in sterile water, dried, and plated onto nutrient agar (NA) and eosin methylene blue agar (EMB) media (3). A similar bacterium was isolated from all samples. After 2 days, white to creamy bacterial colonies on NA and emerald green colonies on EMB developed. Five independent strains were subjected to further biochemical, molecular, and pathogenicity tests. Bacterial strains were gram-negative, motile rods, grew at 37°C, were facultatively anaerobic, oxidase-negative, phosphatase-negative, and catalase-positive. They degraded pectate, were sensitive to erythromycin, did not utilize Keto-methyl glucoside, were indole production-negative, and reduced sugars from sucrose (3). Acid production was negative from sorbitol and arabitol, but positive from melibiose and citrate. PCR amplification of the pel gene by Y1 and Y2 primers produced a 434-bp fragment (2). Amplification of the intergenic transcribed spacer (ITS) region by G1 and L1 primers (4) gave two amplicons ca. 550 and 580 bp long. The expected amplicon was not produced with any of the strains using primers Br1f/L1r and Eca1f/Eca2r (1), whereas a 550-bp PCR product, typical of Pectobacterium carotovorum subsp. carotovorum, was obtained with primers EXPCCF and EXPCCR (1). Based on biochemical and molecular characteristics, and analysis of PCR-RFLP of 16S-ITS-23R rRNA genes using Rsa I enzyme (4), all five bacterial strains were identified as P. carotovorum subsp. carotovorum. BLAST analysis of the 16S rRNA sequence (GenBank Accession No KC189032) showed 100% identity to the 16S rRNA of P. carotovorum subsp. carotovorum strain PPC192. For pathogenicity tests, four mature pepper fruits of cv. Annuum were inoculated by injecting 10 μl of a bacterial suspension (108 CFU/ml) into pericarps and the fruits were incubated in a moist chamber at 80 to 90% relative humidity and 30°C. After 72 h, water-soaked lesions similar to those observed in the fields and greenhouses were observed and bacteria with the same characteristics were consistently reisolated, thereby fulfilling Koch's postulates. Symptoms were not observed on water-inoculated controls. References: (1) S. Baghaee-Ravari et al. Eur. J. Plant Pathol. 129:413, 2001. (2) A. Darraas et al. Appl. Environ. Microbiol. 60:1437, 1994. (3) N. W Schaad et al. Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society Press, St Paul, MN, 2001. (4) I. K. Toth et al. Appl. Environ. Microbiol. 67:4070, 2001.

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