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  1. Chai WL, Moharamzadeh K, Brook IM, Van Noort R
    Biotech Histochem, 2011 Aug;86(4):242-54.
    PMID: 20392135 DOI: 10.3109/10520291003707916
    The success of dental implant treatment depends on the healing of both hard and soft tissues. While osseointegration provides initial success, the biological seal of the peri-implant soft tissue is crucial for maintaining the long term success of implants. Most studies of the biological seal of peri-implant tissues are based on animal or monolayer cell culture models. To understand the mechanisms of soft tissue attachment and the factors affecting the integrity of the soft tissue around the implants, it is essential to obtain good quality histological sections for microscopic examination. The nature of the specimens, however, which consist of both metal implant and soft peri-implant tissues, poses difficulties in preparing the specimens for histomorphometric analysis of the implant-soft tissue interface. We review various methods that have been used for the implant-tissue interface investigation with particular focus on the soft tissue. The different methods are classified and the advantages and limitations of the different techniques are highlighted.
    Matched MeSH terms: Microscopy, Confocal/methods*
  2. Syakila RN, Lim SM, Agatonovic-Kustrin S, Lim FT, Ramasamy K
    Anal Bioanal Chem, 2019 Feb;411(6):1181-1192.
    PMID: 30680424 DOI: 10.1007/s00216-018-1544-2
    The cholesterol-lowering properties of 12 lactic acid bacteria (LAB) in the absence or presence of 0.3% bile salts were assessed and compared quantitatively and qualitatively in vitro. A new, more sensitive and cost-effective high-performance thin-layer chromatography method combined with digital image evaluation of derivatised chromatographic plates was developed and validated to quantify cholesterol in LAB culture media. The performance of the method was compared with that of the o-phthalaldehyde method. For qualitative assessment, assimilated fluorescently tagged cholesterol was visualised by confocal microscopy. All LAB strains exhibited a cholesterol-lowering effect of various degrees (19-59% in the absence and 14-69% in the presence of bile salts). Lactobacillus plantarum LAB12 and Pentosaceus pentosaceus LAB6 were the two best strains of lactobacilli and pediococci. They lowered cholesterol levels by 59% and 54%, respectively, in the absence and by 69% and 58%, respectively, in the presence of bile salts. Confocal microscopy showed that cholesterol was localised at the outermost cell membranes of LAB12 and LAB6. The present findings warrant in-depth in vivo study. Graphical abstract (A) 3D plots based on scan at 525 nm of (B) derivatized HPTLC plate of separated cholesterol and (C) confocal microscopic image showing the localisation of NBD-cholesterol assimilated by LAB.
    Matched MeSH terms: Microscopy, Confocal/methods
  3. Hagiwara Y, Koh JEW, Tan JH, Bhandary SV, Laude A, Ciaccio EJ, et al.
    Comput Methods Programs Biomed, 2018 Oct;165:1-12.
    PMID: 30337064 DOI: 10.1016/j.cmpb.2018.07.012
    BACKGROUND AND OBJECTIVES: Glaucoma is an eye condition which leads to permanent blindness when the disease progresses to an advanced stage. It occurs due to inappropriate intraocular pressure within the eye, resulting in damage to the optic nerve. Glaucoma does not exhibit any symptoms in its nascent stage and thus, it is important to diagnose early to prevent blindness. Fundus photography is widely used by ophthalmologists to assist in diagnosis of glaucoma and is cost-effective.

    METHODS: The morphological features of the disc that is characteristic of glaucoma are clearly seen in the fundus images. However, manual inspection of the acquired fundus images may be prone to inter-observer variation. Therefore, a computer-aided detection (CAD) system is proposed to make an accurate, reliable and fast diagnosis of glaucoma based on the optic nerve features of fundus imaging. In this paper, we reviewed existing techniques to automatically diagnose glaucoma.

    RESULTS: The use of CAD is very effective in the diagnosis of glaucoma and can assist the clinicians to alleviate their workload significantly. We have also discussed the advantages of employing state-of-art techniques, including deep learning (DL), when developing the automated system. The DL methods are effective in glaucoma diagnosis.

    CONCLUSIONS: Novel DL algorithms with big data availability are required to develop a reliable CAD system. Such techniques can be employed to diagnose other eye diseases accurately.

    Matched MeSH terms: Microscopy, Confocal/methods
  4. Hassan T, Thiberville L, Hermant C, Lachkar S, Piton N, Guisier F, et al.
    PLoS One, 2017;12(12):e0189846.
    PMID: 29267317 DOI: 10.1371/journal.pone.0189846
    BACKGROUND: Malignant solitary pulmonary nodules (SPN) have become more prevalent, with upper lobes predilection. Probe-based confocal laser endomicroscopy (pCLE) provides in-vivo imaging of SPN. However, the stiffness of the 1mm confocal probe (AlveoFlex) causes difficult accessibility to the upper lobes. A thinner 600μm probe designed for bile duct exploration (CholangioFlex) has the potential to reach the upper lobes.

    OBJECTIVES: To examine the accessibility of malignant SPNs in all segments of the lungs using either the 0.6mm or 1.4 mm probe and to assess the quality and inter observer interpretation of SPN confocal imaging obtained from either miniprobes.

    METHODS: Radial(r)-EBUS was used to locate and sample the SPN. In-vivo pCLE analysis of the SPN was performed using either CholangioFlex (apical and posterior segments of the upper lobes) or AlveoFlex (other segments) introduced into the guide sheath before sampling. pCLE features were compared between the two probes.

    RESULTS: Fourty-eight patients with malignant SPN were included (NCT01931579). The diagnostic accuracy for lung cancer using r-EBUS coupled with pCLE imaging was 79.2%. All the SPNs were successfully explored with either one of the probes (19 and 29 subjects for CholangioFlex and AlveoFlex, respectively). A specific solid pattern in the SPN was found in 30 pCLE explorations. Comparison between the two probes found no differences in the axial fibers thickness, cell size and specific solid pattern in the nodules. Extra-alveolar microvessel size appeared larger using CholangioFlex suggesting less compression effect. The kappa test for interobserver agreement for the identification of solid pattern was 0.74 (p = 0.001).

    CONCLUSION: This study demonstrates that pCLE imaging of SPNs is achievable in all segments of both lungs using either the 0.6mm or 1.4mm miniprobe.

    Matched MeSH terms: Microscopy, Confocal/methods*
  5. A Talip B, Snelling WJ, Sleator RD, Lowery C, Dooley JSG
    BMC Microbiol, 2018 11 26;18(1):196.
    PMID: 30477427 DOI: 10.1186/s12866-018-1335-0
    BACKGROUND: The field of diagnostics continues to advance rapidly with a variety of novel approaches, mainly dependent upon high technology platforms. Nonetheless much diagnosis, particularly in developing countries, still relies upon traditional methods such as microscopy. Biological material, particularly nucleic acids, on archived glass slides is a potential source of useful information both for diagnostic and epidemiological purposes. There are significant challenges faced when examining archived samples in order that an adequate amount of amplifiable DNA can be obtained. Herein, we describe a model system to detect low numbers of bacterial cells isolated from glass slides using (laser capture microscopy) LCM coupled with PCR amplification of a suitable target.

    RESULTS: Mycobacterium smegmatis was used as a model organism to provide a proof of principle for a method to recover bacteria from a stained sample on a glass slide using a laser capture system. Ziehl-Neelsen (ZN) stained cells were excised and catapulted into tubes. Recovered cells were subjected to DNA extraction and pre-amplified with multiple displacement amplification (MDA). This system allowed a minimum of 30 catapulted cells to be detected following a nested real-time PCR assay, using rpoB specific primers. The combination of MDA and nested real-time PCR resulted in a 30-fold increase in sensitivity for the detection of low numbers of cells isolated using LCM.

    CONCLUSIONS: This study highlights the potential of LCM coupled with MDA as a tool to improve the recovery of amplifiable nucleic acids from archived glass slides. The inclusion of the MDA step was essential to enable downstream amplification. This platform should be broadly applicable to a variety of diagnostic applications and we have used it as a proof of principle with a Mycobacterium sp. model system.

    Matched MeSH terms: Microscopy, Confocal/methods*
  6. Torey A, Sasidharan S
    Eur Rev Med Pharmacol Sci, 2011 Aug;15(8):875-82.
    PMID: 21845797
    Candida (C.) albicans infection in its biofilm mode of growth has taken centre point with the increasing recognition of its role in human infections due to the development of resistance to the commonly used antibiotic or phenotypic adaptation within the biofilm. Hence, in this study the inhibitory effect of methanol extract of Cassia (C.) spectabilis leaves was evaluated against biofilm forming C. albicans.
    Matched MeSH terms: Microscopy, Confocal/methods
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