Hasnan Jaafar

Antioxidant Protective Effect of Honey in Cigarette Smoke-Induced Testicular Damage in Rats

Cigarette smoke (CS) can cause testicular damage and we investigated the possible protective effect of honey against CS-induced testicular damage and oxidative stress in rats. CS exposure (8 min, 3 times daily) and honey supplementation (1.2 g/kg daily) were given for 13 weeks. Rats exposed to CS significantly had smaller seminiferous tubules diameter and epithelial height, lower Leydig cell count and increased percentage of tubules with germ cell loss. CS also produced increased lipid peroxidation (TBARS) and glutathione peroxidase (GPx) activity, as well as reduced total antioxidant status (TAS) and activities of superoxide dismutase (SOD) and catalase (CAT). However, supplementation of honey significantly reduced histological changes and TBARS level, increased TAS level, as well as significantly restored activities of GPx, SOD and CAT in rat testis. These findings may suggest that honey has a protective effect against damage and oxidative stress induced by CS in rat testis.

Newcastle diseases virus strain V4UPM displayed oncolytic ability against experimental human malignant glioma

Abstract Introduction: Newcastle disease virus (NDV) is a virus of paramyxovirus family and lately has been studied for the treatment of cancer in human. In this study, we successfully determined the oncolysis potential of NDV vaccine, V4UPM tested on the human glioblastoma multiform cell line (DBTRG.05MG) and human glioblastoma astrocytoma cell line (U-87MG) in vitro and in vivo. The V4UPM strain is a modified V4 strain developed as thermostable feed pellet vaccine for poultry. Objective: The objectives of this study were mainly to evaluate the cytolytic effect and subsequently determine the brain tumor regression potential induced by this strain in athymic mice model. Methodology and results: V4UPM, the avirulent strain of NDV, was propagate and screened for the cytolytic activity towards DBTRG.05MG and U-87MG using MTT assay. The inhibition concentration 50% (IC50) values by monolayer method measured at hour 72 were 23 and 9 HAU/ml, respectively. Further study was carried out to observe an apoptosis of the infected cells by AO/PI staining and revealed the apoptosis features of the treated cells. Subcutaneous human brain tumors grown on the nude mice were treated by V4UPM at IC80 and complete regression of U-87MG-bearing tumor mice was observed. TUNEL assay analysis of treated tumor tissues from treated mice showed an occurrence of apoptosis. Conclusion: From this study, NDV strain V4UPM inhibits the proliferation of experimental human gliomas in tissue culture and IC80 at 520 HAU V4UPM gives potent effect to induced tumor regression and apoptosis in malignant gliomas.

IGF-1 acts as controlling switch for long-term proliferation and maintenance of EGF/FGF-responsive striatal neural stem cells.

Background: Long-term maintenance of neural stem cells in vitro is crucial for their stage specific roles in neurogenesis. To have an in-depth understanding of optimal conditional microenvironmental niche for long-term maintenance of neural stem cells (NSCs), we imposed different combinatorial treatment of growth factors to EGF/FGF-responsive cells. We hypothesized, that IGF-1-treatment can provide an optimal niche for long-term maintenance and proliferation of EGF/FGF-responsive NSCs. Objective: This study was performed to investigate the cellular morphology and growth of rat embryonic striatal tissue derived-NSCs in long-term culture under the influence of different combinatorial effects of certain growth factors, such as EGF, bFGF, LIF and IGF-1. Methods: The NSCs were harvested and cultured from striatal tissue of 18 days old rat embryos. We have generated neurospheres from these NSCs and cultured them till passage 7 (28 days in vitro) under four different conditional microenvironments: (A) without growth factor, (B) EGF/bFGF, (C) EGF/bFGF/LIF, (D) EGF/bFGF/IGF-1 and (E) EGF/bFGF/LIF/IGF-1. Isolated NSCs were characterised by Immunoflouroscence for nestin expression. The cell growth and proliferation was evaluated at different time intervals (P1, P3, P5 & P7), assessing the metabolic activity based cell proliferation. Apoptosis was studied in each of these groups by In situ cell death assay. Results: Our results demonstrated certain important findings relevant to long-term culture and maintenance of striatal NSC-derived neurospheres. This suggested that IGF-1 can induce enhanced cell proliferation during early stages of neurogenesis, impose long-term maintenance (up to passage 7) to cultured NSCs and enhance survival efficiency in vitro, in the presence of EGF and FGF. Conclusions: Our findings support the hypothesis that the enforcement of IGF-1 treatment to the EGF/FGF-responsive NSCs, can lead to enhanced cell proliferation during early stages of neurogenesis, and an extended life span in vitro. This information will be beneficial for improving future therapeutic implication of NSCs, by addressing improved in vitro production of NSCs.

Effect of Honey on Testicular Functions in Rats Exposed to Cigarette Smoke

Summary Honey is traditionally consumed by the local Malaysian population as a nutrient, as well as for the enhancement of fertility. The decline in male reproductive health and fertility in the last 30 years has been linked to environmental toxicants including cigarette smoke (CS). In human and experimental studies, CS exposure has been associated with decreased plasma testosterone level, lower sperm count and increased percentage of abnormal sperm. The aim of this study, therefore, was to determine the possible protective role of honey against the toxic effects of CS on testicular functions in rats. Thirty-two adult Sprague-Dawley rats were randomly divided into 4 groups (8 rats per group) i.e. control group, honey-treated group (H), cigarette smoke-exposed group (CS) and honey-treated plus CS-exposed group (H+CS). Rats in control and CS groups received oral administration of distilled water daily while rats in H and H+CS groups received honey (1.2 g/kg body weight) orally by gavage daily. Rats in CS and H+CS groups were also exposed to CS for 8 min (3 times/day). After 13 weeks of treatment, each rat was sacrificed for reproductive parameters analysis. Rats in CS group had significantly lower sperm count, daily sperm production, percentage of motile sperm and testosterone level as well as a higher percentage of abnormal sperm compared to control and H groups. However, supplementation of honey significantly improved all these parameters in H+CS group. Administration of honey significantly attenuated the toxic effects of CS on spermatogenesis and testosterone level in rats. This study suggests that honey might have a protective effect against CS-induced impaired testicular functions in rats.

Detection of mycobacterium tuberculosis in Ziehl-Neelsen stained tissue images using Zernike moments and hybrid multilayered perceptron network

Conventional clinical diagnosis of tuberculosis disease such as manual screening by microbiologist are tedious, laborious and time consuming. Therefore, more research has been carried out to develop technologies that able to automate the detection process. This paper presents an automated approach to tuberculosis bacilli detection in tissue section. The proposed approach employs image processing technique and neural network for the segmentation and detection of tuberculosis bacilli. First, images of tuberculosis bacilli in tissue samples are captured using light microscope after stained with Ziehl-Neelsen staining method. Then colour image segmentation using moving k-mean clustering is used to extract tuberculosis bacilli from the tissue image. Two colour spaces, RGB and C-Y colour, were utilised in order to improve the quality of segmentation and robust against various staining condition. Next, geometrical features of Zernike moments are calculated. From these features, the best features that could detect tuberculosis bacilli with higher accuracy were selected using hybrid multilayered perceptron network. Experimental results demonstrate that the proposed method is efficient and accurate to detect the tubercle bacilli in tissue.

Effect of different doses of Malaysian honey on reproductive parameters in adult male rats

The aim of this study was to evaluate the effect of different doses of Malaysian honey on male reproductive parameters in adult rats. Thirty‐two healthy adult male Sprague–Dawley rats were randomly divided into four groups (eight rats per group). Group 1 (control group) was given 0.5 ml of distilled water. Groups 2, 3 and 4 were given 0.2, 1.2 and 2.4 g kg−1 body weight of honey respectively. The rats were treated orally by gavage once daily for 4 weeks. Honey did not significantly alter body and male reproductive organs weights. The rats in Group 3 which received honey at 1.2 g kg−1 had significantly higher epididymal sperm count than those in Groups 1, 2 and 4. No significant differences were found for the percentage of abnormal sperm, elongated spermatid count, reproductive hormonal levels as well as the histology of the testis among the groups. In conclusion, Malaysian honey at a dose of 1.2 g kg−1 daily significantly increased epididymal sperm count without affecting spermatid count and reproductive hormones. These findings might suggest that oral administration of honey at this dose for 4 weeks may enhance spermiogenesis in adult rats.

Exogenous leptin administration decreases sperm count and increases the fraction of abnormal sperm in adult rats

Daily intraperitoneal injection of 5-30 microg/kg body weight of leptin for 42 days to adult rats decreases sperm count and increases the fraction of abnormal sperm.

In human invasive breast ductal carcinoma, tumor stromal macrophages and tumor nest macrophages have distinct relationships with clinicopathological parameters and tumor angiogenesis

Tumor-associated macrophages play a crucial role in breast cancer progression and tumor angiogenesis. However, evaluation of tumor-associated macrophages incorporating their histological locations is lacking. The aim of this study was to clarify whether macrophages in tumor stroma and macrophages in tumor cell nests have distinctive properties in relation to pertinent breast cancer clinicopathological parameters and tumor angiogenesis. In 94 human invasive breast ductal carcinomas, tumor-associated macrophages were immunostained with anti-CD68 antibody and counted or graded according to these histological locations. Microvessels were immunostained with anti-CD34 antibody and counted for microvessel density. We found that the presence of tumor stromal and tumor nest macrophages was closely correlated (p = 0.001). Both tumor stromal and tumor nest macrophages were associated with mitotic count (p = 0.001 and p = 0.037, respectively). However, only higher tumor stromal macrophage grades were associated with higher tumor grades (p = 0.004) and negative estrogen receptor status (p = 0.007). Multivariate analysis showed that tumors with a high mitotic count score (score 3 vs. scores 1 and 2) had a higher tumor stromal macrophage density (Grades III and IV) when adjusted for tumor size, tubule formation, and estrogen receptor status (odds ratio 3.41, p = 0.010). The tumor nest macrophage count significantly correlated with the microvessel density (p < 0.001). These results imply that tumor stromal macrophages and tumor nest macrophages residing in different tumor microenvironments have distinctive roles.

MicroRNA Expression Profile of Neural Progenitor-Like Cells Derived from Rat Bone Marrow Mesenchymal Stem Cells under the Influence of IGF-1, bFGF and EGF

Insulin-like growth factor 1 (IGF-1) enhances cellular proliferation and reduces apoptosis during the early differentiation of bone marrow derived mesenchymal stem cells (BMSCs) into neural progenitor-like cells (NPCs) in the presence of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). BMSCs were differentiated in three groups of growth factors: (A) EGF + bFGF, (B) EGF + bFGF + IGF-1, and (C) without growth factor. To unravel the molecular mechanisms of the NPCs derivation, microarray analysis using GeneChip® miRNA arrays was performed. The profiles were compared among the groups. Annotated microRNA fingerprints (GSE60060) delineated 46 microRNAs temporally up-regulated or down-regulated compared to group C. The expressions of selected microRNAs were validated by real-time PCR. Among the 46 microRNAs, 30 were consistently expressed for minimum of two consecutive time intervals. In Group B, only miR-496 was up-regulated and 12 microRNAs, including the let-7 family, miR-1224, miR-125a-3p, miR-214, miR-22, miR-320, miR-708, and miR-93, were down-regulated. Bioinformatics analysis reveals that some of these microRNAs (miR-22, miR-214, miR-125a-3p, miR-320 and let-7 family) are associated with reduction of apoptosis. Here, we summarize the roles of key microRNAs associated with IGF-1 in the differentiation of BMSCs into NPCs. These findings may provide clues to further our understanding of the mechanisms and roles of microRNAs as key regulators of BMSC-derived NPC maintenance.

Chitosan dermal substitute and chitosan skin substitute contribute to accelerated full-thickness wound healing in irradiated rats.

Wounds with full-thickness skin loss are commonly managed by skin grafting. In the absence of a graft, reepithelialization is imperfect and leads to increased scar formation. Biomaterials can alter wound healing so that it produces more regenerative tissue and fewer scars. This current study use the new chitosan based biomaterial in full-thickness wound with impaired healing on rat model. Wounds were evaluated after being treated with a chitosan dermal substitute, a chitosan skin substitute, or duoderm CGF. Wounds treated with the chitosan skin substitute showed the most re-epithelialization (33.2 ± 2.8%), longest epithelial tongue (1.62 ± 0.13 mm), and shortest migratory tongue distance (7.11 ± 0.25 mm). The scar size of wounds treated with the chitosan dermal substitute (0.13 ± 0.02 cm) and chitosan skin substitute (0.16 ± 0.05 cm) were significantly decreased (P < 0.05) compared with duoderm (0.45 ± 0.11 cm). Human leukocyte antigen (HLA) expression on days 7, 14, and 21 revealed the presence of human hair follicle stem cells and fibroblasts that were incorporated into and surviving in the irradiated wound. We have proven that a chitosan dermal substitute and chitosan skin substitute are suitable for wound healing in full-thickness wounds that are impaired due to radiation.