The in vivo biocompatibility and toxicity of PVA/NOCC scaffold were tested by comparing them with those of a biocompatible inert material HAM in a rat model. On Day 5, changes in the blood parameters of the PVA/NOCC-implanted rats were significantly higher than those of the control. The levels of potassium, creatinine, total protein, A/G, hemoglobulin, erythrocytes, WBC, and platelets were not significantly altered in the HAM-implanted rats, when compared with those in the control. On Day 10, an increase in potassium, urea, and GGT levels and a decrease in ALP, platelet, and eosinophil levels were noted in the PVA/NOCC-implanted rats, when compared with control. These changes were almost similar to those noted in the HAM-implanted rats, except for the unaltered potassium and increased neutrophil levels. On Day 15, the total protein, A/G, lymphocyte, monocyte, and eosinophil levels remained unaltered in the PVA/NOCC-implanted rats, whereas urea, A/G, WBC, lymphocyte, and monocyte levels remained unchanged in the HAM-implanted rats. Histology and immunohistochemistry analyses revealed inflammatory infiltration in the PVA/NOCC-implanted rats, but not in the HAM-implanted rats. Although a low toxic tissue response was observed in the PVA/NOCC-implanted rats, further studies are necessary to justify the use of this material in tissue engineering applications.
PURPOSE: The aim was to assess the morphology of the Asian corneal endothelium, particularly the index of polymegethism and its relationship to endothelial cell density assessments.
METHODS: The central regions of the corneal endothelium of 72 healthy, normal weight, non-contact lens wearers were assessed by non-contact specular microscopy. There were six groups of persons, aged between 18 and 27 years, whose ethnic origins were self-reported as Asian (Mainland China, Hong Kong, India, Malaysia, Pakistan or other Asian groups). Single images including at least 100 contiguous cells were obtained from each subject and were processed by planimetry after manual border marking to minimise errors. Calculations were made of the coefficient of variation of cell area (COV; polymegethism) as well as the endothelial cell density and percentage of six-sided cells.
RESULTS: The COV values ranged from 18 to 30.4 per cent (group mean 24.5 ± 2.8 per cent). The corneas had essentially normal thickness (0.530 ± 0.026 mm), with the endothelia showing a high percentage of six-sided cells (66.3 ± 7.1 per cent) and none showing even mild signs of polymegethism. Endothelial cell density estimates based on average cell areas ranged from 2,020 to 3,623 cells per mm(2) and from 2,157 to 3,835 with calculations based on individual cell areas. The difference between the two estimates of endothelial cell density was strongly related to the coefficient of variation (r = 0.765).
CONCLUSIONS: This result is consistent with several older reports indicating that it is uncommon to find polymegethism in healthy young adult Asian individuals. Nonetheless, even low coefficient of variation values can substantially affect the estimations of the cell density.
KEYWORDS: Asian; cornea; endothelium; human; polymegathism
As a natural anti-oxidant source, Tualang honey, produced by wild bees nesting on the Tualang tree (Koompassia excelsa) is expected to have positive influence on health, including memory. This study investigated the effect of Tualang honey on the cell count of memory formation related hippocampal pyramidal neuron and on spatial memory performance (SMP) of rats using the radial arm maze (RAM). Sprague Dawley male rats (n=24), 7-8 weeks old were divided into two groups; experimental group group force-fed 1 mL/100 g body weight with 70% honey (HG); and the control group with 0.9% saline (CG) for 12 weeks. Nissl staining technique (with cresyl violet) was employed for neurohistological analysis of the hippocampal tissue. Six randomly selected rats from each group were used for the neuronal soma counting of pyramidal cell layer CA1, CA3a and CA3c regions. Two-way ANOVA analysis showed positively significant differences between treatment and control groups for SMP comparison of working memory and reference memory components, as well as the number of pyramidal neurons. Hence, this positive effects of Tualang honey, as demonstrated behaviorally and neurohistologically, supported report that Tualang honey could improve memory and deter hippocampal morphological impairments; possibly due to its high anti-oxidant properties.
We studied by scanning electron microscopy the effects of the repeated use of a newly-designed, disposable, suction trephine (Microsurgical, Ltd, Bedfordshire, England) on the nature of the cut of successive corneal buttons from sheep eyes. After the trephine was used more than nine times, the cut edge became more irregular as successive buttons were produced. Also, progressively greater degrees of endothelial cells adjacent to the cut edge were lost. Although the corneal buttons initially were circular, those produced after the 20th cut became more oval. These results suggest that a disposable corneal trephine should be reused no more than nine times.
The effect of physical shearing on the attachment of six Pseudomonas aeruginosa strains and six Burkholderia cepacia strains to glass, stainless steel, polystyrene and Teflon® was determined. A significant (p < 0.05) decrease in hydrophobicity was apparent for all P. aeruginosa strains (17-36%) and B. cepacia, MS 5 (20%) after shearing. A significant (p < 0.05) decrease in attachment of some P. aeruginosa (0.2-0.5 log CFU/cm2) and B. cepacia (0.2-0.4 log CFU/cm2) strains to some surface types was apparent after shearing. Significant (p < 0.05) correlation was observed for both numbers of flagellated cells and hydrophobicity against attachment to glass, stainless steel and polystyrene for P. aeruginosa while only hydrophobicity showed significant correlation against the same surfaces for B. cepacia. Scanning electron microscopy and protein analysis showed that shearing removed surface proteins from the cells and may have led to the observed changes in hydrophobicity and attachment to abiotic surfaces.
This study was aimed to investigate the effects of dietary fatty acids on the accretion pattern of major fat pads, inguinal fat cellularity, and their relation with plasma leptin concentration. Forty Sprague-Dawley rats were randomly assigned into four groups and received the following diets for 22 weeks: (1) standard rat chow diet (CTRL), (2) CTRL + 10% (w/w) butter (HFAR), (3) CTRL + 3.33% (w/w) menhaden fish oil + 6.67% (w/w) soybean oil (MFAR), and (4) CTRL + 6.67% (w/w) menhaden fish oil + 3.33% (w/w) soybean oil (LFAR). Inguinal fat cellularity and plasma leptin concentration were measured in this study. Results for inguinal fat cellularity showed that the mean adipocyte number for the MFAR (9.2 ∗ 10⁵ ± 3.6) and LFAR (8.5 ∗ 10⁵ ± 5.1) groups was significantly higher (P < 0.05) than the rest, while the mean adipocyte diameter of HFAR group was larger (P < 0.05) (46.2 ± 2.8) than the rest. The plasma leptin concentration in the HFAR group was higher (P < 0.05) (3.22 ± 0.32 ng/mL), than the other groups. The higher inguinal fat cellularity clearly indicated the ability of the polyunsaturated fatty acids (PUFA) and butter supplemented diets to induce hyperplasia and hypertrophy of fat cells, respectively, which caused adipocyte remodeling due to hyperleptinemia.
The performance of pipeline system used in petroleum industry is crucially declined by natural microbial activities and
demanding extra operational cost. Requirement on high capability of functional substances is attracting worldwide
research interest. The aim of this paper was to study the effectiveness of benzyltriethylammonium chloride (BTC) on
reducing the activity of a consortium bacteria consisting of sulfate-reducing bacteria (C-SRB). C-SRB was isolated from
tropical crude oil and enumeration of this consortium was measured by viable cell count technique. The effectiveness of
BTC was calculated from potentiodynamic polarization method and biofilm analysis was performed by scanning electron
microscope. The viable cell count technique indicated that the maximum growth of C-SRB was approximately 160 trillion
CFU/mL at 7 days incubation period. BTC was capable of reducing biocorrosion activity due to adsorption process and
mitigating SRB species. Biofilm analysis has proven that C-SRB activity is minimized due to less presence of bacterial
growth, extracellular polymeric substances and corrosion product. In conclusion, BTC is capable to inhibit C-SRB activity
on biocorrosion of carbon steel pipeline.
Ficus deltoidea has been shown to possess antioxidant properties that could prevent the development of chronic inflammatory bone diseases. In this study, the efficacy of F. deltoidea in preventing alveolar bone resorption in osteoporotic rats induced by ovariectomy (OVX) was investigated. Twenty-four female Wistar rats were divided into four groups (n = 6) consisting of sham-operated (SO), ovariectomized control (OVXN), ovariectomized treated with estrogen (OVXP), and ovariectomized treated with F. deltoidea extract (OVXF). At the beginning of the study, two nonovariectomized, healthy rats were sacrificed to serve as baseline (BL). Treatment of the rats commenced two weeks after ovariectomy-the OVXP rats that served as positive control received Premarin® (64.5 μg/kg body weight), while OVXF rats were given F. deltoidea (800 mg/kg body weight); both agents were administered orally for two months. The negative control group of rats (OVXN) and the SO group received deionized water, also administered via oral gavage. At necropsy, morphometric assessment of the interradicular bone of the first molar was carried out using a micro-CT scanner, while quantification of osteoclasts and osteoblasts was performed histologically. The results showed that no statistically significant differences among the groups (p > 0.05) for bone morphometric assessment. However, trabecular thickness in the OVXF group was similar to BL, while trabecular separation and alveolar bone loss height were lower than those of the OVXN group. Histologically, the OVXF group demonstrated a significantly lower number of osteoclasts and a higher number of osteoblasts compared with OVXN (p=0.008 and p=0.019, respectively; p < 0.05). In conclusion, F. deltoidea has the capacity to prevent alveolar bone loss in ovariectomy-induced osteoporosis rats by potentially preserving trabecular bone microarchitecture and to decrease osteoclast and increase osteoblast cell count.
To compare the potential retinal toxicity of two commercially Brilliant blue G dyes (Brilliant Peel and Ocublue Plus) and Indocyanine green (ICG) at usual clinical concentration.
A 22-year-old Indian female was referred to Sg Buloh hospital with the diagnosis of bilateral keratoconus. On examination, slit lamp biomicroscopy and corneal topography revealed stage 3 keratoconus in the right eye and stage 2 keratoconus in the left eye. Corneal cell morphology in both eyes was evaluated using confocal microscope. In qualitative observation, almost all corneal layers in right eye except endothelium were partially or completely obscured by haze. Additionally, morphological alterations, such as elongation of keratocyte nuclei and cluster of cells, and dark bands in the anterior stroma were observed in right eye. In the left eye, the amount of haze was less, allowing better visibility of the corneal layers compared with the right eye. The dark bands were evident in the posterior stroma. Quantitative analysis showed that anterior and posterior stromal keratocyte density and endothelium cell density were relatively low in the right eye (834.0, 700.5, and 2,133 cells/mm2, respectively) compared with the left eye (934.1, 750.6, and 2,361 cells/mm2, respectively). In this case, the right eye, exhibiting stage 3 keratoconus, showed more morphological alteration, particularly in the anterior stroma compared with the left eye with stage 2 keratoconus. Increased severity of the disease can explain these differences in corneal cell morphology.
Blood samples were collected from 24 immature male, 55 immature female and 99 mature female water buffalo kept at an experimental farm in the Northern Territory. Haematological analysis was performed on blood collected in dipotassium--ethylene diamine tetra acetic acid while biochemical analysis was performed on serum and plasma (for glucose) samples. Haematological values of mature buffalo were similar to those recorded for swamp buffalo in Malaysia. Blood cell appearances were similar to those reported for adult Indian river buffalo though values recorded for red cell components were higher. Statistical analysis revealed no significant differences between immature male and female buffalo. Red cell components, eosinophils, total plasma and serum proteins, albumin, gamma globulins, inorganic phosphate and the enzyme gamma-glutamyl transferase were significantly higher for mature female buffalo when compared to immature females. Reasons for the differences were not fully determined but the effect of age and nutritional status in combination with a variable period of domestication were considered.
Agricultural development is a major threat to global biodiversity, and effective conservation actions are crucial. Physiological repercussions of life alongside human-modified landscapes can undermine adaptable species' health and population viability; however, baseline data are lacking for many wildlife species. We assessed the physiological status of a generalist carnivore, the Malay civet (Viverra tangalunga), persisting within an extensively human-modified system in Sabah, Malaysian Borneo. We characterized hematology and serum biochemistry panels from civets sampled across a mosaic landscape comprising tropical forest fragments and oil palm plantations. Intra-population variation in certain blood parameters were explained by expected biological drivers such as sex, age category and sampling season. Furthermore, we determined several erythrocyte measures, immune cell counts and dietary biochemistry markers significantly varied with proximity to oil palm plantation boundaries. These findings were supported by a case study, whereby blood profiles of GPS collared male civets were contrasted based on their exclusive use of forests or use of oil palm plantations. These data provide robust and valuable first insights into this species' physiological status and suggest agricultural landscapes are impacting the persisting population.
The exponential fed-batch cultivation of Bacillus sphaericus UPMB10 in 2 l stirred tank fermenter was performed by feeding the initial batch culture with 14 g l-1 of glycerol according to the algorithm aimed at controlling the specific growth rate (μ) of the bacterium. Very high viable cell count (1.14 x 1010 cfu ml-1), which was four times higher as compared to batch cultivation, was achieved in the fed-batch with a controlled μ at 0.4 h-1. In repeated exponential fed-batch cultivation, consisting of four cycles of harvesting and recharging, a final cell concentration of 1.9 x 1011 cfu ml-1 was obtained at the end of the fourth cycle (46 h). Meanwhile, acetylene reduction of cell samples collected from repeated fed-batch cultivation remained unchanged and was maintained at around 20 nmol C2H2 h-1 ml-1 after prolonged cultivation period, and was comparable to those obtained in batch and exponential fed-batch cultivation. Glycerol could be used as a carbon source for high performance cultivation of B. sphaericus, a nitrogen fixing bacterium, in repeated fed-batch cultivation with high cell yield and cell productivity. The productivity (0.68 g l-1 h-1) for repeated fed-batch cultivation increased about 6 times compared to that obtained in conventional batch cultivation (0.11 g l1 h-1). A innovative method in utilizing glycerol for efficient cultivation of nitrogen fixing bacterium could be beneficial to get more understanding and reference in manipulating the integrated plans for sustainable and profitable biodiesel industry.
Chitosan nanoparticles (CSNPs) have potential applications in stem cell research. In this study, ex vivo cytotoxicity of CSNPs on mouse bone marrow-derived (MBMCs) hematopoietic stem and progenitor cells (HSPCs) was determined. MBMCs were exposed to CSNPs of different particle sizes at various concentrations for up to 72 h. Cytotoxicity effect of CSNPs on MBMCs was determined using MTT, Live/Dead Viability/Cytotoxicity assays and flow cytometry analysis of surface antigens on HSCs (Sca-1(+)), myeloid-committed progenitors (CD11b(+), Gr-1(+)), and lymphoid-committed progenitors (CD45(+), CD3e(+)). At 24 h incubation, MBMCs' viability was not affected by CSNPs. At 48 and 72 h, significant reduction was detected at higher CSNPs concentrations. Small CSNPs (200 nm) significantly reduced MBMCs' viability while medium-sized particle (∼400 nm) selectively promoted MBMCs growth. Surface antigen assessment demonstrated lineage-dependent effect. Significant decrease in Sca-1(+) cells percentage was observed for medium-sized particle at the lowest CSNPs concentration. Meanwhile, reduction of CD11b(+) and Gr-1(+) cells percentage was detected at high and intermediate concentrations of medium-sized and large CSNPs. Percentage of CD45(+) and CD3e(+) cells along with ROS levels were not significantly affected by CSNPs. In conclusion, medium-sized and large CSNPs were relatively non-toxic at lower concentrations. However, further investigations are necessary for therapeutic applications.
Leukaemic stem cells have heterogenous differentiation potential. The immunophenotypes of blast cells are usually consistent throughout the disease course even at relapse. Rarely, blast cells may undergo a ‘lineage switch’ during the course of disease especially during relapse. We would like to highlight such a case in a 10- year old boy who presented with a two weeks history of lethargy, poor appetite, low grade fever, respiratory distress, cardiac failure, generalized oedema and hepatosplenomegaly. Full blood count showed a leucocyte count of 41.5x10 9 /L and platelet count of 37x10 9 /L. The peripheral blood film showed presence of numerous blast cells. Bone marrow aspiration revealed a hypercellular marrow, which consisted of mainly blast cells with high nuclear to cytoplasmic ratio and inconspicuous nucleoli. Immunophenotyping and cytochemistry results were consistent with the diagnosis of Tcell acute lymphoblastic leukaemia. The patient achieved remission after treatment with UK ALL 97 protocol, regime B chemotherapy. However, he relapsed seven months after the initial diagnosis with 26% blast cells in the bone marrow aspirate. The majority was L1 blast cells admixed with some L2 blast cells. Immunophenotyping was consistent with common precursor B acute lymphoblastic leukaemia. The treatment was changed to a more lineage specific chemotherapy. Nonetheless, the patient never achieved remission and was planned for palliative management. This case illustrated a unique and rare case of rapid lineage switch from T-cell acute lymphoblastic leukaemia to common precursor B-cell acute lymphoblastic leukaemia.
Introduction: Virgin coconut oil (VCO) is known for its health and therapeutic benefits. However, the immunomodulatory effects of VCO have not been extensively investigated. Objective: The present study was devoted to examining the effects of VCO on cyclophosphamide (CY)-induced toxicity of lymphoid tissues. Methods: Thirty healthy male Wistar rats were sorted into 5 groups of 6 animals. The first control (NC) group was given distilled water via gavage at 5 ml/kg once daily. The second (CY) group received CY orally at 10 mg/kg/day for 4 weeks. Rats in the other three groups (CV5, CV10, and CV15) were given 10 mg/kg/day CY for 4 weeks, 5 m/kg/day, 10 ml/kg/day and 15 ml/kg/day VCO for 6 weeks, respectively. Rats were sacrificed at the end of 6th week; blood sample from the animals was collected for full blood count and biochemical analysis. The thymus and spleen of each animal was processed for histological examination. Results: The thymus and spleen showed marked reduction in lymphoid cellularity following daily administration of CY. The thymus also showed a marked reduction in the size of the medulla, and the white pulp areas of spleen had reduction in the follicle number and size. Supplementation with 10 ml/kg and 15 ml/kg VCO showed evidence of restoration of both the thymus and splenic lymphoid architecture. The total white cell counts, absolute lymphocyte counts and plasma globulin levels of the VCO groups were significantly increased compared to CY group. Conclusion: VCO displayed potential protective effects on CYinduced histological changes in lymphoid tissues.
Background Eosinophilic chronic rhinosinusitis (eCRS) is linked with skewed T-helper 2 or immunoglobulin E (IgE)-mediated allergic responses, with differing diagnosis, prognosis, and management to non-eCRS. Objective The association between biomarkers and eCRS was investigated to assess the predictors of eCRS. Methods A cross-sectional study of adult patients with chronic rhinosinusitis (CRS) undergoing endoscopic sinus surgery was conducted. eCRS was defined by histopathological assessment showing >10 eosinophils/high-power field on sinus mucosal biopsy. Blood tests were performed preoperatively and assessed for a full blood count including eosinophils and a white cell count (WCC) as well as biochemical markers of inflammation and atopy including Immunoglobulin E (IgE), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and ImmunoCAP testing for serum-specific IgE. Comparisons between eCRS and non-eCRS patients were performed. Results 345 patients (48.1% female, age 48.72 ± 15.06 years) were recruited, with 206 (59.7%) identified as eCRS, 41% with asthma and 47% CRS with nasal polyps. eCRS patients were more likely to have asthma ( P 0.24 × 109/L), eosinophil ratio (>4.27% of total WCC), and lower ESR when compared with non-eCRS.
The purification of human adipose-derived stem cells (hADSCs) from human adipose tissue cells (stromal vascular fraction) was investigated using membrane filtration through poly(lactide-co-glycolic acid)/silk screen hybrid membranes. Membrane filtration methods are attractive in regenerative medicine because they reduce the time required to purify hADSCs (i.e., less than 30 min) compared with conventional culture methods, which require 5-12 days. hADSCs expressing the mesenchymal stem cell markers CD44, CD73, and CD90 were concentrated in the permeation solution from the hybrid membranes. Expression of the surface markers CD44, CD73, and CD99 on the cells in the permeation solution from the hybrid membranes, which were obtained using 18 mL of feed solution containing 50 × 10⁴ cells, was statistically significantly higher than that of the primary adipose tissue cells, indicating that the hADSCs can be purified in the permeation solution by the membrane filtration method. Cells expressing the stem cell-associated marker CD34 could be successfully isolated in the permeation solution, whereas CD34⁺ cells could not be purified by the conventional culture method. The hADSCs in the permeation solution demonstrated a superior capacity for osteogenic differentiation based on their alkali phosphatase activity, their osterix gene expression, and the results of mineralization analysis by Alizarin Red S and von Kossa staining compared with the cells from the suspension of human adipose tissue. These results suggest that the hADSCs capable of osteogenic differentiation preferentially permeate through the hybrid membranes.
Apoptosis is a tightly programmed cell suicide which occurs in multiple physiologic and pathological conditions where it plays an important role in tissue development and homeostasis by eliminating unwanted and damaged cells. Appropriate apoptosis signalling is crucial in maintaining the fine balance between cell death and cell survival in cancer. In response to death stimuli the morphology of the cell undergoes unique changes. The aim of this study was to examine and compare the changes in the cell surface morphology using scanning electron microscopy in HCS-2 cells, following 24 hour treatment with components of highly active antiretroviral therapy (HAART) at their clinical plasma concentrations. The cells were fixed in 2.5% Glutaraldehyde and post-fixed in 1% osmium tetroxide. The cells were then dehydrated through a graded series of alcohol and treated with hexamethyl-disilazane, then coated with a double layer of carbon. The cells were viewed under a Zeiss Ultra FEG Scanning Electron Microscope and a one way ANOVA and Tukey Kramer Post Hoc test was conducted based on the scoring of surface morphology of the cells using JMP 11 statistical software. The drugs used in this study induced morphological features which are known to be characteristic of apoptotic cell death. The drug combinations (ATP and LPV/r) were seemingly more effective than individual treatments in inducing cell death because morphological features observed were more advanced than those observed in individual treatments. However, LPV/r was more potent than ATP. In conclusion, HAART showed anticancer properties by inducing cell death through apoptosis.