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Isolation and characterization of Enterococcus faecium DSM 20477 with ability to secrete antimicrobial substance for the inhibition of oral pathogen Streptococcus mutans UKMCC 1019

Ng ZJ, Zarin MA, Lee CK, Phapugrangkul P, Tan JS

Arch Oral Biol, 2020 Feb;110:104617.
PMID: 31794906 DOI: 10.1016/j.archoralbio.2019.104617

Streptococcus mutans and Candida albicans are the main oral pathogens which contribute to dental caries that affects all ages of human being.
OBJECTIVES: This study focuses on the potential of crude cell free supernatant (CCFS) from lactic acid bacteria (LAB) to inhibit of the growth of S. mutans UKMCC 1019.
DESIGN: A total of 61 CCFS from LAB strains were screened for their inhibitory ability against S. mutans UKMCC 1019 by broth microdilution method. The selected LAB with highest antimicrobial activity was identified and its CCFS was characterized for pH stability, temperature tolerance, enzyme sensitivity, metabolism of carbohydrates, enzymatic activities and antimicrobial activity against S. mutans UKMCC 1019 and C. albicans UKMCC 3001 by well diffusion assay. The effect of CCFS on cell structure of S. mutans UKMCC 1019 was observed under transmission electron microscopy (TEM).
RESULTS: The CCFS from isolate CC2 from Kimchi showed the highest inhibition against S. mutans UKMCC 1019, which was 76.46 % or 4406.08 mm2/mL and it was identified to be most closely related to Enterococcus faecium DSM 20477 based on 16 s rRNA sequencing. The CCFS of E. faecium DSM 20477 had high tolerance to acidic and alkaline environment as well as high temperature. It also shows high antifungal activities against C. albicans UKMCC 3001 with 2362.56 mm2/mL. Under TEM, the cell walls and the cytoplasm membrane of S. mutans UKMCC 1019 were disrupted by the antimicrobial substance, causing cell lysis.
CONCLUSIONS: Hence, the CCFS from E. faecium DSM 20477 is a potential bacteriocin in future for the treatment of dental caries.

Matched MeSH terms: Cytoplasm
In-vitro phagocytosis and intracellular killing of Pasteurella multocida B:2 by phagocytic cells of buffaloes

Puspitasari Y, Annas S, Adza-Rina MN, Zamri-Saad M

Microb Pathog, 2019 Jun;131:170-174.
PMID: 30978429 DOI: 10.1016/j.micpath.2019.04.012

Pasteurella multocida B:2 is a Gram-negative organism causing haemorrhagic septicaemia (HS) in buffaloes. It causes severe pulmonary infection, leading to infiltration of numerous macrophages and neutrophils. Despite the inflammatory response, buffaloes succumb to HS. This study aims to evaluate the in-vitro efficacy of macrophages and neutrophils of buffalo following exposure to P. multocida B:2. In-vitro infections were done using 107 cfu/ml of P. multocida B:2 for Group 1, Escherichia coli for Group 2 and Mannhaemia haemolytica A:2 for Group 3 cells. The inoculated cell cultures were harvested at 0, 30, 60 and 120 min post-exposure and the phagocytic, killing and cell death rates were determined. Both phagocytosis and killing rates of all bacteria increased over time. Phagocytosis involved between 71% and 73% neutrophils and between 60% and 64% macrophages at 120 min. Killing rate of all bacteria involved between 76% and 79% for neutrophils and between 70% and 74% for macrophages at 120 min. Death rate of neutrophils ranged between 67% in Group 3, and 88% in Group 1 at 120 min, significantly (p  0.05) than Group 2. Similar pattern was observed for death rate of macrophages. The phagocytosis and killing rates of P. multocida B:2 were similar to other bacterial species used in this study but more neutrophils and macrophages were dead following infection by P. multocida B:2 than M. haemolytica A:2.

Matched MeSH terms: Cytoplasm/microbiology; Cytoplasm/physiology*
Fulltext Immunocytochemical localization of carbonic anhydrase in the pseudobranch tissue of the rainbow trout Oncorhynchus mykiss

Rahim SM, Mazlan AG, Simon KD, Delaunoy JP, Laurent P

J Zhejiang Univ Sci B, 2014 Feb;15(2):194-200.
PMID: 24510712 DOI: 10.1631/jzus.B1200297

Pseudobranch function has long interested scientists, but its role has yet to be elucidated. Several studies have suggested that pseudobranchs serve respiratory, osmoregulatory, and sensory functions. This work investigated the immunolocalization of pseudobranch carbonic anhydrase (CA) in the teleost fish species rainbow trout (Oncorhynchus mykiss) to clarify its physiological function. CA was purified from rainbow trout gills O. mykiss and specific antibodies were raised. Immunoblotting between tissue homogenates of pseudobranch and gill CA antibodies showed specific immunostaining with only one band corresponding to CA in the pseudobranch homogenate. Results of immunohistochemical technique revealed that CA was distributed within pseudobranch cells and more precisely in the apical parts (anti-vascular) of cells. The basal (vascular) parts of cells, tubular system, blood capillaries, and pillar cells were not immunostained. Immunocytochemistry confirmed these results and showed that some CA enzyme was cytoplasmic and the remainder was linked to membranous structures. The results also showed that the lacunar tissue layers did not display immunoperoxidase activity. Our results indicated that pseudobranch CA may have a function related to the extracellular medium wherein CA intervenes with the mechanism of stimulation of afferent nerve fibers.

Matched MeSH terms: Cytoplasm/metabolism
Design, mechanism, delivery and therapeutics of canonical and Dicer-substrate siRNA

Raja MAG, Katas H, Amjad MW

Asian J Pharm Sci, 2019 Sep;14(5):497-510.
PMID: 32104477 DOI: 10.1016/j.ajps.2018.12.005

Upon the discovery of RNA interference (RNAi), canonical small interfering RNA (siRNA) has been recognized to trigger sequence-specific gene silencing. Despite the benefits of siRNAs as potential new drugs, there are obstacles still to be overcome, including off-target effects and immune stimulation. More recently, Dicer substrate siRNA (DsiRNA) has been introduced as an alternative to siRNA. Similarly, it also is proving to be potent and target-specific, while rendering less immune stimulation. DsiRNA is 25-30 nucleotides in length, and is further cleaved and processed by the Dicer enzyme. As with siRNA, it is crucial to design and develop a stable, safe, and efficient system for the delivery of DsiRNA into the cytoplasm of targeted cells. Several polymeric nanoparticle systems have been well established to load DsiRNA for in vitro and in vivo delivery, thereby overcoming a major hurdle in the therapeutic uses of DsiRNA. The present review focuses on a comparison of siRNA and DsiRNA on the basis of their design, mechanism, in vitro and in vivo delivery, and therapeutics.

Matched MeSH terms: Cytoplasm
Fulltext Electrocautery resection as a treatment option for canine transmissible venereal tumour (CTVT)

Rajendren, S. K., Krishnan, K., Ganesh, T. N., Roslan, N. S., Hashim, N. A., Mohamad, M. A.

Jurnal Veterinar Malaysia, 2019;31(2):19-22.
MyJurnal

A 5-year-old Mongrel was brought presented with the complaint of having serosanguineous discharge from penis for a month since adoption. Physical examination revealed cauliflower-like mass at the bulbus glandis. Presence of numerous anisokaryotic and anisocytotic round to oval histiocytes with multivacuolated cytoplasm from cytology, an evidence of canine transmissibale venereal tumour (CTVT). The mass was successfully surgically resected using electrocautery and was in remission for 12 months (since January 2019).

Matched MeSH terms: Cytoplasm
Fulltext Secondary Sea-Blue Histiocytosis in a Patient with Transfusion Dependent HbE-Beta Thalassaemia and Osteosarcoma

Saad Eldeen Bakheet O, Yusof N, Raja Zahratul A, Ithnin A, Abdul Aziz S, Alias H

Indian J Hematol Blood Transfus, 2016 Jun;32(Suppl 1):262-6.
PMID: 27408409 DOI: 10.1007/s12288-015-0582-6

Secondary sea-blue histiocytosis occurs more frequently than the primary form and occurs consequent to a wide range of metabolic and haematologic disorders including thalassaemia. We report an 18-year-old Chinese boy with transfusion-dependent HbE-beta thalassaemia who complained of pain and swelling at the left iliac crest region for 2 months duration. Physical examination revealed pallor with hepatosplenomegaly. Local examination revealed a huge swelling 12 cm × 12 cm in diameter, firm in consistency and tender. Histopathological examination of the mass revealed an osteosarcoma. His bone marrow aspirate showed numerous sea-blue histiocytes, the cytoplasm of which was closely packed with fine granules that stained blue with May-Grunwald-Giemsa. The nuclei were centrally located in some cells and displaced towards the periphery in other cells. There was no malignant cell infiltration in the marrow. The case is reported due to the co-incidental dual pathology in our patient (HbE-beta thalassaemia and osteosarcoma) and the unusual bone marrow finding of numerous sea-blue histiocytes.

Matched MeSH terms: Cytoplasm
Fulltext Quantification of intracellular payload release from polymersome nanoparticles

Scarpa E, Bailey JL, Janeczek AA, Stumpf PS, Johnston AH, Oreffo RO, et al.

Sci Rep, 2016 07 11;6:29460.
PMID: 27404770 DOI: 10.1038/srep29460

Polymersome nanoparticles (PMs) are attractive candidates for spatio-temporal controlled delivery of therapeutic agents. Although many studies have addressed cellular uptake of solid nanoparticles, there is very little data available on intracellular release of molecules encapsulated in membranous carriers, such as polymersomes. Here, we addressed this by developing a quantitative assay based on the hydrophilic dye, fluorescein. Fluorescein was encapsulated stably in PMs of mean diameter 85 nm, with minimal leakage after sustained dialysis. No fluorescence was detectable from fluorescein PMs, indicating quenching. Following incubation of L929 cells with fluorescein PMs, there was a gradual increase in intracellular fluorescence, indicating PM disruption and cytosolic release of fluorescein. By combining absorbance measurements with flow cytometry, we quantified the real-time intracellular release of a fluorescein at a single-cell resolution. We found that 173 ± 38 polymersomes released their payload per cell, with significant heterogeneity in uptake, despite controlled synchronisation of cell cycle. This novel method for quantification of the release of compounds from nanoparticles provides fundamental information on cellular uptake of nanoparticle-encapsulated compounds. It also illustrates the stochastic nature of population distribution in homogeneous cell populations, a factor that must be taken into account in clinical use of this technology.

Matched MeSH terms: Cytoplasm
Fulltext Hyalinizing clear cell carcinoma of the soft palate - a case report

Sharifah Intan Safuraa, Sethu Subha, Muhamad Doi, Sellymiah Adzman

Malaysian Journal of Medicine and Health Sciences, 2020;16(1):333-336.
MyJurnal

Hyalinizing clear cell carcinoma presents as a painless submucosal mass commonly located at the palate and base of tongue. It is a rare tumour and has often been misdiagnosed for other more common tumours with clear cytoplasm, such as acinic cell carcinoma, clear cell oncocytoma or mucoepidermoid carcinoma. HCCC has been reported as a low grade malignant tumour with a high rate of cervical metastases. Due to its rarity, there is no treatment protocol. However, the treatment of choice is wide local excision and the neck disease is treated with neck dissection or ra- diotherapy or both with no conclusive outcome as incidence is too low or underreported with no long term follow up. Our case highlights the diagnosis difficulties in such rare cases, and the need for longer follow up post excision to determine outcome and recurrence rates.

Matched MeSH terms: Cytoplasm
Podoplanin, E-cadherin, β-catenin, and CD44v6 in recurrent ameloblastoma: their distribution patterns and relevance

Siar CH, Ishak I, Ng KH

J Oral Pathol Med, 2015 Jan;44(1):51-8.
PMID: 25059841 DOI: 10.1111/jop.12203

Ameloblastoma is a benign but locally infiltrative odontogenic epithelial neoplasm with a high risk for recurrence. Podoplanin, a lymphatic endothelium marker, putatively promotes collective cell migration and invasiveness in this neoplasm. However, its role in the recurrent ameloblastoma (RA) remains unclear. As morphological, signaling, and genetic differences may exist between primary and recurrent tumors, clarification of their distribution patterns is of relevance.

Matched MeSH terms: Cytoplasm/chemistry
Differential expression of transcription factors Snail, Slug, SIP1, and Twist in ameloblastoma

Siar CH, Ng KH

J Oral Pathol Med, 2014 Jan;43(1):45-52.
PMID: 23560539 DOI: 10.1111/jop.12065

Epithelial-to-mesenchymal transition (EMT) via the mechanism of transcription repression is a crucial process for the induction of invasiveness in many human tumors. Ameloblastoma is a benign odontogenic epithelial neoplasm with a locally infiltrative behavior. Twist, an EMT promoter, has been implicated in its invasiveness. The roles of the other transcription factors remain unclarified.

Matched MeSH terms: Cytoplasm/ultrastructure
Claudin expression and tight junction protein localization in the lining epithelium of the keratocystic odontogenic tumors, dentigerous cysts, and radicular cysts

Siar CH, Abbas SA

Oral Surg Oral Med Oral Pathol Oral Radiol, 2013 May;115(5):652-9.
PMID: 23601220 DOI: 10.1016/j.oooo.2013.02.013

The aim of this study was to evaluate the expression and localization of tight junction proteins (TJPs) or claudins in the keratocystic odontogenic tumor (KCOT) and to correlate with its biological behavior.

Matched MeSH terms: Cytoplasm/pathology; Cytoplasmic Granules/ultrastructure
Differential expression of Notch receptors and their ligands in desmoplastic ameloblastoma

Siar CH, Nakano K, Han PP, Nagatsuka H, Ng KH, Kawakami T

J Oral Pathol Med, 2010 Aug 1;39(7):552-8.
PMID: 20337864 DOI: 10.1111/j.1600-0714.2009.00871.x

In mammals, the Notch gene family encodes four receptors (Notch1-4), and all of them are important for cell fate decisions. Notch signaling pathway plays an essential role in tooth development. The ameloblastoma, a benign odontogenic epithelial neoplasm, histologically recapitulates the enamel organ at bell stage. Notch has been detected in the plexiform and follicular ameloblastoma. Its activity in the desmoplastic ameloblastoma is unknown.

Matched MeSH terms: Cytoplasm/ultrastructure
Denture hyperplasia and oncocytic metaplasia

Siar CH, Ng KH, Ngui CH

Ann Dent, 1992;51(1):27-8.
PMID: 1632623

A case of denture hyperplasia of the upper labial sulcus with concomitant oncocytic metaplastic changes is described. The patient concerned is an elderly male wearing an ill-fitting upper full denture.

Matched MeSH terms: Cytoplasm/ultrastructure; Cytoplasmic Granules/ultrastructure
Peripheral odontogenic fibroma with clear cell odontogenic epithelium

Siar CH, NG KH

J N Z Soc Periodontol, 1995.
PMID: 9227094

The clinical and histological features of the peripheral odontogenic fibroma are briefly outlined. A case arising from the attached lingual gingiva between the mandibular right permanent first molar and the second molar in a 67 year old Indian female is reported here. The unusual occurrence of marked clear cell differentiation within the odontogenic epithelial component, and histogenetic link to the clear cell rests of the dental lamina and surface epithelium are discussed.

Matched MeSH terms: Cytoplasm/pathology
Immunohistochemical study of the distribution of S-100 protein in pleomorphic adenomas of minor salivary glands

Siar CH, Ng KH

J Nihon Univ Sch Dent, 1992 Jun;34(2):96-105.
PMID: 1380072

Twelve pleomorphic adenomas of minor salivary gland origin were examined for the distribution of S-100 protein, detected using the peroxidase-antiperoxidase (PAP) method. Strong S-100 protein immunoreactivity was noted in areas containing plasmacytoid cells, stellate and spindle cells against a myxochondroid or hyalinous stroma, and solid epithelial areas. Tubular and duct-like structures showed variable stainability. Stromal tissue and normal salivary glands were generally negative for S-100 protein. These findings were compared with those reported elsewhere.

Matched MeSH terms: Cytoplasm/ultrastructure; Cytoplasm/chemistry
Plexiform granular cell odontogenic tumor: unicystic variant

Siar CH, Ng KH, Jalil NA

Oral Surg. Oral Med. Oral Pathol., 1991 Jul;72(1):82-5.
PMID: 1891247

Plexiform granular cell odontogenic tumor of the mandible has recently been described. The cardinal histopathologic feature, as its name suggests, is a monophasic plexiform pattern of granular cells; the principal tumor in the differential diagnosis is granular cell ameloblastoma. Unlike the two previously reported cases of plexiform granular cell odontogenic tumor, which occurred as solid tumors in elderly men, the lesion reported here is a unicystic variant occurring in a middle-aged woman.

Matched MeSH terms: Cytoplasm/ultrastructure
Fulltext Patched-1 and smoothened, a hedgehog receptor and signal transducer are highly expressed in diffuse large B-cell lymphoma

Siti Nur Lina Azman, Huzlinda Hussin, Salmiah Md Said, Zanariah Alias, Maizaton Atmadini Abdullah

Malaysian Journal of Medicine and Health Sciences, 2017;13(2):1-6.
MyJurnal

Introduction: The Hedgehog (Hh) signalling pathway is a developmental signalling pathway involved in normal mammalian developmental and homeostasis of adult renewable tissues. In most adult tissues, this pathway remains silent and previous studies have shown that constitutive activation of Hedgehog signalling pathway leads to various types of malignancies including medulloblastomas, basal cell carcinoma, gastrointestinal, breast and prostate cancer. The purpose of this study was to investigate the immunohistochemical expression of Hedgehog pathway proteins in Diffuse Large B-cell Lymphoma and determine their association with overall survival (OS). Methods: Positive control using normal tonsils were included in each batch of immunohistochemical staining procedure. Results: PTCH1 proteins were highly expressed in DLBCL and showed strong staining intensity in 107 (100%) cases and SMO proteins were expressed in 105 (98.1%) cases. PTCH1 proteins were localised in the nucleus of tumour cells, whereas SMO proteins were mainly localised in the cytoplasm of tumour cells. Positive expression of PTCH1 and SMO proteins and overall survival of DLBCL patients were correlated with age, gender, race and tumour location. There was no significant correlation between the expression of these two proteins with any of the parameters. PTCH1 expression showed significant association with SMO expression (P=0.03). Conclusions: Our findings suggest that high expression of both PTCH1 and SMO may be important in the pathogenesis of DLBCL. However, additional mechanisms that may contribute to the activation of HH signalling in DLBCL needs to be further explored.

Matched MeSH terms: Cytoplasm
Fulltext In silico identification of novel tuberculosis drug targets in mycobacterium tuberculosis P450 enzymes by interaction study with azole drugs

Suresh Kumar

Malaysian Journal of Medicine and Health Sciences, 2020;16(101):24-30.
MyJurnal

Introduction: Tuberculosis (TB) is one of the utmost serious infectious diseases worldwide. The emergence of multi- drug resistance demands the development of better or new putative drug targets for tuberculosis. Recent studies sug- gest Mycobacterium tuberculosis cytochrome P450 enzymes as promising drug targets and azole drugs as potential inhibitors. Methods: Various computational tools, like Expasy Protparam, Swiss model, RaptorX and Phyre2 were used to analyze 12 Mycobacterium tuberculosis P450 enzymes and determine their three-dimensional structure. The structural validation was done through a Ramachandran plot using RAMPAGE server. The docking of P450 enzymes with azole drugs was done with autodock ver 4.2.6. Results: Based on sub-cellular localization prediction using CEL- LO tool, P450 enzymes CYP123A1, CYP132A1, CYP135A1, CYP136A1, CYP140A1, and CYP143A1 were predicted to be in the cytoplasm. Through structure assessment by Ramachandran plot, the best homology modelled proteins were docked with azole drugs like clotrimazole, croconazole, econazole, fluconazole, itraconazole, itraconazole, ketaconazole and micronazole by using autodock. By docking method it is identified that ketaconazole drug has a high affinity towards most of the mycobacterium P450 enzymes followed by the itrconazole drug. CYP123A1 enzyme is preferable as a drug target due to high binding affinity towards ketoconazole followed by CYP135A1, CYP140A1 enzymes. Conclusion: This study would help in identifying putative novel drug targets in Mycobacterium tuberculosis, which can lead to promising candidates for the optimization and development of novel anti-mycobac- terial agents.

Matched MeSH terms: Cytoplasm
Fulltext Sciatic nerve repair with tissue engineered nerve: Olfactory ensheathing cells seeded poly(lactic-co-glygolic acid) conduit in an animal model

Tan CW, Ng MH, Ohnmar H, Lokanathan Y, Nur-Hidayah H, Roohi SA, et al.

Indian J Orthop, 2013 Nov;47(6):547-52.
PMID: 24379458 DOI: 10.4103/0019-5413.121572

BACKGROUND AND AIM: Synthetic nerve conduits have been sought for repair of nerve defects as the autologous nerve grafts causes donor site morbidity and possess other drawbacks. Many strategies have been investigated to improve nerve regeneration through synthetic nerve guided conduits. Olfactory ensheathing cells (OECs) that share both Schwann cell and astrocytic characteristics have been shown to promote axonal regeneration after transplantation. The present study was driven by the hypothesis that tissue-engineered poly(lactic-co-glycolic acid) (PLGA) seeded with OECs would improve peripheral nerve regeneration in a long sciatic nerve defect.
MATERIALS AND METHODS: Sciatic nerve gap of 15 mm was created in six adult female Sprague-Dawley rats and implanted with PLGA seeded with OECs. The nerve regeneration was assessed electrophysiologically at 2, 4 and 6 weeks following implantation. Histopathological examination, scanning electron microscopic (SEM) examination and immunohistochemical analysis were performed at the end of the study.
RESULTS: Nerve conduction studies revealed a significant improvement of nerve conduction velocities whereby the mean nerve conduction velocity increases from 4.2 ΁ 0.4 m/s at week 2 to 27.3 ΁ 5.7 m/s at week 6 post-implantation (P < 0.0001). Histological analysis revealed presence of spindle-shaped cells. Immunohistochemical analysis further demonstrated the expression of S100 protein in both cell nucleus and the cytoplasm in these cells, hence confirming their Schwann-cell-like property. Under SEM, these cells were found to be actively secreting extracellular matrix.
CONCLUSION: Tissue-engineered PLGA conduit seeded with OECs provided a permissive environment to facilitate nerve regeneration in a small animal model.

Matched MeSH terms: Cytoplasm
Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase

Tay SP, Cheong SK, Hamidah NH, Ainoon O

Malays J Pathol, 1999 Jun;21(1):37-43.
PMID: 10879277

A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.

Matched MeSH terms: Cytoplasm/enzymology*

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