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Fulltext HPV positivity and its' influencing factors among invasive cervical cancer women in Malaysia

Sharifa Ezat Wan Puteh, Norin Rahayu Samsuddin, Sharifah Noor Akmal Syed Hussain, Shamsul Azhar Shah, Syed Mohamed Aljunid

Int J Public Health Res, 2011;1(1):13-22.
MyJurnal

Accepted 10 August 2011.
Introduction Cervical cancer (CC) is the second most prevalent female cancer in Malaysia. Almost 70% of its’ causal factors are attributable to oncogenic human papillomavirus (HPV) types 16, 18 and other risk factors. HPV genotypes distributions are also noted to differ by geographical area.
Methods This was cross sectional study conducted in 2007, to determine the influencing factors of HPV positivity and prevalence of HPV infections among patients with cervical cancer in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). Patients’ paraffin-embedded cervical tissues kept in the Pathology Department from 1999 to 2007 were randomly selected. A total of 81 medical records with complete information were chosen as samples and patients were contacted for consent. Tissue samples were further derived for PCR DNA for HPV genotyping. Analyses included descriptive statistics; bivariate χ2 test and correlation were used to determine relationship between factors and HPV positivity. Significance level of less than 0.05 was taken as statistically significant.
Results Mean age of cancer diagnosis was at 52 ± 12.2 years. Women of Chinese ethnicity was the highest ethnicity to be HPV positive at 65.4% and squamous cell carcinoma was more commonly found (59.3%) compared with other types of cancers. The prevalence of HPV positivity was 92.6% with type 16 being the most common (74.1%), followed by type 33 (30.9%) and 18 (22.2%). Multiple HPV infections were a common finding at 54.3%. Factors thought to influence positivity i.e. age of intercourse, number of sexual partners, number of parity, smoking status of patients and their partners, oral contraceptive usage, presence of chronic illnesses and cancer stage were not significantly associated with HPV positivity. Increased CC severity level was not associated with increased number of HPV infections (Pearson correlation 0.58; p =0.607).
Conclusions High HPV positivity at 92.6% was found among ICC patients. Factors thought to influence HPV positivity were not significant. The top three HPV genotypes were type 16 followed by type 33 and 18. However, local women HPV serotypes findings need to be replicated in a larger population sample.

Matched MeSH terms: Paraffin
Fulltext Fluid-solid phase transition of n-alkane mixtures: Coarse-grained molecular dynamics simulations and diffusion-ordered spectroscopy nuclear magnetic resonance

Shahruddin S, Jiménez-Serratos G, Britovsek GJP, Matar OK, Müller EA

Sci Rep, 2019 Jan 30;9(1):1002.
PMID: 30700804 DOI: 10.1038/s41598-018-37799-7

Wax appearance temperature (WAT), defined as the temperature at which the first solid paraffin crystal appears in a crude oil, is one of the key flow assurance indicators in the oil industry. Although there are several commonly-used experimental techniques to determine WAT, none provides unambiguous molecular-level information to characterize the phase transition between the homogeneous fluid and the underlying solid phase. Molecular Dynamics (MD) simulations employing the statistical associating fluid theory (SAFT) force field are used to interrogate the incipient solidification states of models for long-chain alkanes cooled from a melt to an arrested state. We monitor the phase change of pure long chain n-alkanes: tetracosane (C24H50) and triacontane (C30H62), and an 8-component surrogate n-alkane mixture (C12-C33) built upon the compositional information of a waxy crude. Comparison to Diffusion Ordered Spectroscopy Nuclear Magnetic Resonance (DOSY NMR) results allows the assessment of the limitations of the coarse-grained models proposed. We show that upon approach to freezing, the heavier components restrict their motion first while the lighter ones retain their mobility and help fluidize the mixture. We further demonstrate that upon sub-cooling of long n-alkane fluids and mixtures, a discontinuity arises in the slope of the self-diffusion coefficient with decreasing temperature, which can be employed as a marker for the appearance of an arrested state commensurate with conventional WAT measurements.

Matched MeSH terms: Paraffin
Fulltext Fleet enema: a straight up method of management in tar burn injuries

Farrah-Hani, I., Mohamed Sam-Aan, H., K., Enda, Srijit D.

Medicine & Health, 2020;15(2):321-326.
MyJurnal

Lecuran tar, walaupun dikelaskan sebagai kecederaan kimia, disebabkan oleh perbezaan mekanisma kecederaan dan kesukaran untuk mengeluarkan agen daripada kulit tanpa menyebabkan kerosakan kulit dengan lebih lanjutnya, merupakan satu masalah yang sukar untuk diselesaikan. Terutamanya, apabila pesakit pediatrik terlibat. Lecuran tar biasanya merupakan kecederaan pekerjaan dan kebanyakannya berlaku kepada lelaki dewasa. Dua kes dibentangkan di sini, seorang budak lelaki berusia enam tahun dan seorang gadis berusia 15 tahun yang mengalami luka lecuhan tar dan berjaya dirawat dengan menggunakan enema minyak mineral Fleet® dan paraffin tulle Bactigras®, yang mudah didapati di kebanyakan hospital.

Matched MeSH terms: Paraffin
First Report of Lasiodiplodia theobromae Causing Stem Canker of Jatropha curcas in Malaysia

Sulaiman R, Thanarajoo SS, Kadir J, Vadamalai G

Plant Dis, 2012 May;96(5):767.
PMID: 30727556 DOI: 10.1094/PDIS-06-11-0482-PDN

Physic nut (Jatropha curcas L.) is an important biofuel crop worldwide. Although it has been reported to be resistant to pests and diseases (1), stem cankers have been observed on this plant at several locations in Peninsular Malaysia since early February 2008. Necrotic lesions on branches appear as scars with vascular discoloration in the tissue below the lesion. The affected area is brownish and sunken in appearance. Disease incidence of these symptomatic nonwoody plants can reach up to 80% in a plantation. Forty-eight samples of symptomatic branches collected from six locations (University Farm, Setiu, Gemenceh, Pulau Carey, Port Dickson, and Kuala Selangor) were surface sterilized in 10% bleach, rinsed twice with sterile distilled water, air dried on filter paper, and plated on water agar. After 4 days, fungal colonies on the agar were transferred to potato dextrose agar (PDA) and incubated at 25°C. Twenty-seven single-spore fungal cultures obtained from all locations produced white, aerial mycelium that became dull gray after a week in culture. Pycnidia from 30-day-old pure cultures produced dark brown, oval conidia that were two celled, thin walled, and oval shape with longitudinal striations. The average size of the conidia was 23.63 × 12.72 μm with a length/width ratio of 1.86. On the basis of conidial morphology, these cultures were identified as Lasiodiplodia theobromae. To confirm the identity of the isolates, the internal transcribed spacer (ITS) region was amplified with ITS1/ITS4 primers and sequenced. The sequences were deposited in GenBank (Accession Nos. HM466951, HM466953, HM466957, GU228527, HM466959, and GU219983). Sequences from the 27 isolates were 99 to 100% identical to two L. theobromae accessions in GenBank (Nos. HM008598 and HM999905). Hence, both morphological and molecular characteristics confirmed the isolates as L. theobromae. Pathogenicity tests were performed in the glasshouse with 2-month-old J. curcas seedlings. Each plant was wound inoculated by removing the bark on a branch to a depth of 2 mm with a 10-mm cork borer. Inoculation was conducted by inserting a 10-mm-diameter PDA plug of mycelium into the wound and wrapping the inoculation site with wetted, cotton wool and Parafilm. Control plants were treated with plugs of sterile PDA. Each isolate had four replicates and two controls. After 6 days of incubation, all inoculated plants produced sunken, necrotic lesions with vascular discoloration. Leaves were wilted and yellow above the point of inoculation on branches. The control plants remained symptomless. The pathogen was successfully reisolated from lesions on inoculated branches. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts and is common in tropical and subtropical regions of the world (2,3). To our knowledge, this is the first report of stem canker associated with L. theobromae on J. curcas in Malaysia. References: (1) S. Chitra and S. K. Dhyani. Curr. Sci. 91:162, 2006. (2) S. Mohali et al. For. Pathol. 35:385, 2005. (3) E. Punithalingam. Page 519 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, Surrey, UK. 1976.

Matched MeSH terms: Paraffin
First Report of Fusarium Wilt Caused by Fusarium oxysporum on Roselle in the United States

Ploetz RC, Palmateer AJ, Geiser DM, Juba JH

Plant Dis, 2007 May;91(5):639.
PMID: 30780734 DOI: 10.1094/PDIS-91-5-0639A

Roselle, Hibiscus sabdariffa var. sabdariffa, is an annual that is grown primarily for its inflated calyx, which is used for drinks and jellies. It is native from India to Malaysia, but was taken at an early date to Africa and is now widely grown in the tropics and subtropics (2). In late 2005, dying plants were noted by a producer in South Florida. Plants wilted, became chlorotic, and developed generally unthrifty, sparse canopies. Internally, conspicuous vascular discoloration was evident in these plants from the roots into the canopy. After 5 days on one-half-strength potato dextrose agar (PDA), salmon-colored fungal colonies grew almost exclusively from surface-disinfested 5 mm2 pieces of vascular tissue. On banana leaf agar, single-spored strains produced the following microscopic characters of Fusarium oxysporum: copious microconidia on monophialides, infrequent falcate macroconidia, and terminal and intercalary chlamydospores. Partial, elongation factor 1-α (EF1-α) sequences were generated for two of the strains, O-2424 and O-2425, and compared with previously reported sequences for the gene (3). Maximum parsimony analysis of sequences showed that both strains fell in a large, previously described clade of the F. oxysporum complex (FOC) that contained strains from agricultural hosts, as well as human clinical specimens (2; clade 3 in Fig. 4); many of the strains in this clade have identical EF1-α sequences. Strains of F. oxysporum recovered from wilted roselle in Egypt, O-647 and O-648 in the Fusarium Research Center collection, were distantly related to the Florida strains. We are not aware of other strains of F. oxysporum from roselle in other international culture collections. Roselle seedlings were inoculated with O-2424 and O-2425 by placing a mycelial plug (5 mm2, PDA) over a small incision 5 cm above the soil line and then covering the site with Parafilm. Parafilm was removed after 1 week, and plants were incubated under ambient temperatures (20 to 32°C) in full sun for an additional 5 weeks (experiment 1) or 7 weeks (experiment 2). Compared with mock-inoculated (wound + Parafilm) control plants, both O-2424 and O-2425 caused significant (P < 0.05) vascular disease (linear extension of discolored xylem above and below wound site) and wilting (subjective 1 to 5 scale); both isolates were recovered from affected plants. F. oxysporum-induced wilt of roselle has been reported in Nigeria (1) and Malaysia (4) where the subspecific epithet f. sp. rosellae was used for the pathogen. We are not aware of reports of this disease elsewhere. To our knowledge, this is the first report of F. oxysporum-induced wilt of roselle in the United States. Research to determine whether the closely related strains in clade 3 of the FOC are generalist plant pathogens (i.e., not formae speciales) is warranted. References: (1) N. A. Amusa et al. Plant Pathol. J. 4:122, 2005. (2) J. Morton. Pages 81-286 in: Fruits of Warm Climates. Creative Resource Systems, Inc., Winterville, NC, 1987. (3) K. O'Donnell et al. J. Clin. Microbiol. 42:5109, 2004. (4) K. H. Ooi and B. Salleh. Biotropia 12:31, 1999.

Matched MeSH terms: Paraffin
First Report of Bark Dieback on Blueberry Caused by Botryosphaeria dothidea in Korea

Choi IY

Plant Dis, 2011 Feb;95(2):227.
PMID: 30743439 DOI: 10.1094/PDIS-05-10-0371

This study was conducted to identify the causal organism of bark dieback disease of highbush blueberry (Vaccinium corymbosum L.) observed in Korea. Blueberry, a woody plant that is native to North America, belongs to the family Ericaceae and genus Vaccinium. Of the 400 species of blueberry in the world, most are distributed in the tropics of Malaysia and Southeast Asia. Highbush blueberry is abundantly grown in Canada and the United States and has become a popular commercial crop in Korea for products such as jam, wine, and sauce. Bark dieback disease of blueberry was found in Sunchang (<5% incidence), Jeollabuk-do, Korea in July 2009. Typical symptoms of the disease were blight and dieback on the stems with lesions extending along entire branches. Morphological examination revealed that the perithecia were of the globose type with a nipple, 155 to 490 (374.6) μm, and brown on the dead bark. Asci were bitunicate and clavate or cylindrical with dimensions of 63 to 125 × 16 to 20 μm and containing eight ascospores. Ascospores were of the long ovoid type with dimensions of 13.2 to 23.7 (17.98) × 25.4 to 41.1 (33.21) μm. From extracted genomic DNA, the internal transcribed spacer (ITS)-5.8S ribosomal DNA region was amplified with universal primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). A BLAST search of GenBank with the ITS sequence revealed that the Sunchang isolate (GenBank Accession No. HQ384217) had 99 to 100% sequence identity with the following Botryosphaeria dothidea accessions: FJ517657, AJ938005, FJ478129, FJ171723, and AJ938004. Phylogenetic analysis with the Sunchang isolate, B. dothidea strains, and related species revealed that the B. dothidea isolate and strains comprised a monophyletic group distinguished from other Botryosphaeria spp. including B. ribis, B. parva, B. protearum, B. lutea, B. australis, B. rhodina, B. obtuse, and B. stevensii (2). On the basis of morphological and molecular results, the isolate was identified as B. dothidea (Moug.) Ces. & De Not. A culture of B. dothidea isolate was grown on potato dextrose agar (PDA) for 10 days. A 5-mm plug was inoculated into stem wounds created with a No. 2 cork borer in 20 2-year-old disease-free blueberry plants grown in a greenhouse. Six plants inoculated with only PDA plugs served as noninoculated controls. The wounds were covered with Parafilm. After 3 months, the Parafilm was removed and black lesions were observed at the fungal inoculation sites, while no lesion was observed on the control plants. To complete Koch's postulates, the fungus was reisolated from the lesions and confirmed to be B. Dothidea (1). There is an urgent need to determine the spread of this disease in Korea, estimate the losses, and develop methods for reducing damage through biological and eco-friendly cultural control methods. References: (1) D. Jurc et al. Plant Pathol. 55:299, 2006. (2) B. Slippers et al. Mycologia 96:83, 2004.

Matched MeSH terms: Paraffin
Fulltext Feasibility of T-cell receptor gamma (TCRgamma) gene rearrangement on formalin-fixed, paraffin-embedded tissues by PCR assays

Tai YC, Peh SC

Singapore Med J, 2003 May;44(5):250-5.
PMID: 13677361

T- and B-lymphocytes are involved in recognition of foreign antigen by the specificity of their surface T-cell receptor and immunoglobulin, generated by gene rearrangement. Each T- and B-lymphocyte carries unique rearranged TCR or immunoglobulin gene, which has been applied to detect clonal from non-clonal T- and B-cell proliferation.

Matched MeSH terms: Paraffin Embedding
Fulltext Fast Melt Cocoa Butter Tablet: Effect of Waxes, Starch, and PEG 6000 on Physical Properties of the Preparation

Liew KB, Ming LC, Goh BH, Peh KK

Molecules, 2022 May 13;27(10).
PMID: 35630605 DOI: 10.3390/molecules27103128

A fast melt tablet (FMT) is well regarded as an alternative delivery system that might help resolve a patient's non-compliance issue. The main objective of this study was to develop a cocoa butter-based FMT. Additives, namely 5-15% of PEG 6000, beeswax, paraffin wax, and corn starch, were incorporated into the cocoa butter-based FMT to study the effects of these additives with the physical characteristic of a cocoa butter FMT. An optimum-based formulation was chosen according to the desired hardness and disintegration time and the taste masking property achieved with the model drug-dapoxetine. The analysis demonstrated that incorporating beeswax (15%) and paraffin wax (15%) could prolong the disintegration time by at least two-fold. On the contrary, the presence of corn starch was found to cause an increase in the hardness and reduction of the disintegration time. The disintegration mechanism might be presumed due to the synergistic effect of starch swelling and cocoa butter melting. The hardness value and in vitro disintegration time of the optimum formulation were recorded at 2.93 ± 0.22 kg and 151.67 ± 6.98 s. In terms of dissolution, 80% of dapoxetine was released within 30 min and the dissolution profile was comparable to the innovator product. The formulation was palatable and stable for at least 1 year. The exposure of the FMT formulation at 30 °C for 12 months was reported to be stable. Along with the sound palatability profile and high drug load capacity, the current formulation possesses the desired characteristics to be scaled up and marketed.

Matched MeSH terms: Paraffin*
Fulltext Fabrication and Evaluation of Transdermal Microneedles for a Recombinant Human Keratinocyte Growth Factor

Chellathurai MS, Ling VWT, Palanirajan VK

Turk J Pharm Sci, 2021 Feb 25;18(1):96-103.
PMID: 33634684 DOI: 10.4274/tjps.galenos.2020.21033

Objectives: Microneedle transdermal patches are a combination of hypodermic needles and transdermal patches used to overcome the individual limitations of both injections and patches. The objective of this study was to design a minimally invasive, biodegradable polymeric recombinant human keratinocyte growth factor (rHuKGF) microneedle array and evaluate the prepared biodegradable microneedles using in vitro techniques.
Materials and Methods: Biodegradable polymeric microneedle arrays were fabricated out of poly lactic-co-glycolic acid (PLGA) using the micromolding technique under aseptic conditions, and the morphology of the microneedles was characterized using light microscopy. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to rule out drug-polymer interactions. Standard procedures were used to analyze the prepared microneedle arrays for in vitro drug release and to perform a microneedle insertion test. Enzyme-linked immunosorbent assay was used to quantify rHuKGF.
Results: The PLGA polymer was safe for use in the fabrication of rHuKGF microneedles as there was no interaction between the drug and the polymer. The fabricated rHuKGF microneedle arrays had fully formed microneedles with a height of 600 µm and a base of 300 µm. The drug from the microneedle patch was released in vitro within 30 minutes. The strength of the microneedles in the patch was good, as they were able to reach a depth of 381±3.56 µm into parafilm without any structural change or fracture.
Conclusion: Microneedle transdermal patches were successfully prepared for rHuKGF, and their evaluation suggested excellent quality and uniformity of patch characteristics. This can have potential applications in the therapeutic arena, offering advantages in terms of reduced dosing frequency, improved patient compliance, and bioavailability.

Matched MeSH terms: Paraffin
FISH analysis using PPAR γ-specific probes for detection of PAX8-PPAR γ translocation in follicular thyroid neoplasms

Sharifah NA, Zakaria Z, Chia WK

Methods Mol Biol, 2013;952:187-96.
PMID: 23100233 DOI: 10.1007/978-1-62703-155-4_13

Fluorescence in situ hybridization (FISH) is increasingly gaining importance in clinical diagnostics settings. Due to the ability of the technique to detect chromosomal abnormalities in samples with low cellularity or containing a mixed population of cells even at a single-cell level, it has become more popular in cancer research and diagnosis. Here, we describe the FISH technique for detection of PAX8-PPARγ translocation in follicular thyroid neoplasms, and the optimal protocol for the detection of this fusion gene using in archival formalin-fixed paraffin-embedded (FFPE) thyroid tissue sections.

Matched MeSH terms: Paraffin Embedding
Expression of IGFBP-rP1 in ovarian and breast cancers in association with diabetes mellitus status

Mohd Nafi SN, Siti Azrin AH, Mat Zin AA, Othman NH, Che Jalil NA

Malays J Pathol, 2019 Apr;41(1):33-39.
PMID: 31025635

INTRODUCTION: Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) is an important component of the IGF system that regulates insulin resistance-related to tumour development. The aim of this study is to investigate the expression of IGFBP-rP1 among female cancer patients who are known or not known to have Type 2 Diabetes Mellitus (T2DM).
MATERIALS AND METHODS: Using a cross-sectional design, cases of ovarian and breast cancer with clinical status of T2DM were selected over a 10-year period in Hospital Universiti Sains Malaysia. Immunohistochemical staining for IGFBP-rP1 was performed on paraffin-embedded tissues and the results were correlated with the patient's demographic and clinicopathological data.
RESULTS: A total of 152 breast cancer patients were recruited into the current study with 33.5% (51/152) patients were positive T2DM. Most of the breast cancer patients with T2DM were IGFBP-rP1-negative (66.7%, 34/51). The IGFBP-rP1 expression was significantly difference between breast cancer subjects with and without T2DM (p<0.001). There was no significant association of IGFBP-rP1 expression with data on the demographic and clinicopathological profiles of patients with breast cancer. Meanwhile, positive IGFBP-rP1 expression was evident in 44 out of 108 (40.74%) ovarian cancer cases. Among these cases, 36 were T2DM. In contrast to breast cancer cases, IGFBP-rP1 was mostly expressed among ovarian cancer patients with T2DM (66.7%, 24/36, p < 0.001). However, the -positive expression was not significantly associated with any sociodemographic and clinicopathological features of ovarian cancers.
CONCLUSIONS: Majority of breast cancer patients with T2DM did not express IGFBP-rP1. In contrast, majority of the ovarian cancer patients with T2DM expressed IGFBP-rP1.

Matched MeSH terms: Paraffin Embedding
Fulltext Exploring Two Protocols of FISH Using Cytocell SYT-SSX Probe on Formalin Fixed Paraffin Embedded Tissue Sections

Yajid AI, Mohd Nafi SN, Salehan NA, Tuan Sharif SE

Asian Pac J Cancer Prev, 2020 May 01;21(5):1241-1245.
PMID: 32458628 DOI: 10.31557/APJCP.2020.21.5.1241

BACKGROUND: Chromosomal translocation t(X;18)(p11.2;q11.2) is the cytogenetic hallmark of synovial sarcoma and have been identified as an alternative diagnostic strategy in differentiating synovial sarcoma from other histologic mimics. This study was carried out to test the efficacy of two FISH protocols using the SYT-SSX break apart probe from Cytocell.
METHODOLOGY: Representative paraffin blocks of synovial sarcoma were utilized in this study. FISH study was performed on formalin-fixed paraffin embedded tissue sections using the SYT-SSX break apart probe from Cytocell, to detect two form of SYT-SSX transcript, SYT-SSX1 and SYT-SSX2. FISH protocol, including the hybridization was done following two different protocols, Cytocell FISH protocol and Optimized Dako FISH protocol.
RESULTS: Tissue samples subjected to FISH using Cytocell FISH protocol showed the absence of signal corresponding to the probe used. Utilizing Optimized Dako FISH protocol, the two signals (red and green) corresponding to the break-apart probes was detected. These findings suggested that Optimised Dako FISH protocol is more suited for use with the tested probe on paraffin embedded tissues in comparison to Cytocell FISH protocol.
CONCLUSION: Optimised Dako FISH protocol was noted to be more suited for detecting SYT-SSX FISH signals on paraffin embedded tissues in comparison to Cytocell FISH protocol.

Matched MeSH terms: Paraffin; Paraffin Embedding
Fulltext Evaluation of Seegene Anyplex II Performance for Detection of Human Papillomavirus Genotypes in Formalin-Fixed, Paraffin-Embedded Cervical Cancer Specimens

Haqshenas G, Molano M, Phillips S, Balgovind P, Garland SM, Hawkes D, et al.

Arch Pathol Lab Med, 2024 Mar 01;148(3):353-358.
PMID: 37226838 DOI: 10.5858/arpa.2022-0317-OA

CONTEXT.—: Detection of human papillomavirus (HPV) in formalin-fixed, paraffin-embedded (FFPE) tissues may identify the cause of lesions and has value for the development of new diagnostic assays and epidemiologic studies. Seegene Anyplex II assays are widely used for HPV screening, but their performance using FFPE samples has not been fully explored.
OBJECTIVE.—: To validate Anyplex II HPV HR Detection (Anyplex II, Seegene) using FFPE samples.
DESIGN.—: We used 248 stored DNA extracts from cervical cancer FFPE samples collected during 2005-2015 that tested HPV positive using the RHA kit HPV SPF10-LiPA25, v1 (SPF10, Labo Biomedical Products) HPV genotyping assay, manufacturer-validated for FFPE samples.
RESULTS.—: Of the selected 248 samples, 243 were used in our analysis. Consistent with SPF10 genotyping results, Anyplex II detected all 12 oncogenic types and had an overall HPV detection rate of 86.4% (210 of 243 samples). Anyplex II and SPF10 showed very high agreement for the detection of the 2 most important oncogenic genotypes: HPV 16 (219 of 226; 96.9%; 95% CI, 93.7-98.75) and HPV 18 (221 of 226; 97.8%; 95% CI, 94.9-99.3).
CONCLUSIONS.—: Overall results showed that both platforms produced comparable HPV genotyping results, indicating the suitability of Anyplex II for FFPE samples. The Anyplex II assay has the added convenience of being an efficient, single-well semiquantitative polymerase chain reaction assay. Further optimization of Anyplex II may enhance its performance using FFPE samples by improving the detection limit.

Matched MeSH terms: Paraffin Embedding/methods
Fulltext Evaluation of DNA and RNA extraction methods

Edwin Shiaw CS, Shiran MS, Cheah YK, Tan GC, Sabariah AR

Med J Malaysia, 2010 Jun;65(2):133-7.
PMID: 23756798 MyJurnal

This study was done to evaluate various DNA and RNA extractions from archival FFPE tissues. A total of 30 FFPE blocks from the years of 2004 to 2006 were assessed with each modified and adapted method. Extraction protocols evaluated include the modified enzymatic extraction method (Method A), Chelex-100 extraction method (Method B), heat-induced retrieval in alkaline solution extraction method (Methods C and D) and one commercial FFPE DNA Extraction kit (Qiagen, Crawley, UK). For RNA extraction, 2 extraction protocols were evaluated including the enzymatic extraction method (Method 1), and Chelex-100 RNA extraction method (Method 2). Results show that the modified enzymatic extraction method (Method A) is an efficient DNA extraction protocol, while for RNA extraction, the enzymatic method (Method 1) and the Chelex-100 RNA extraction method (Method 2) are equally efficient RNA extraction protocols.

Matched MeSH terms: Paraffin Embedding*
Endocan expression in placenta of women with hypertension

Chew BS, Ghazali R, Othman H, Ismail NAM, Othman AS, Laim NMST, et al.

J Obstet Gynaecol Res, 2018 Oct 10.
PMID: 30306675 DOI: 10.1111/jog.13836

AIM: The aim of our study was to determine the endocan-1 expression in placenta of hypertensive women, and its association with maternal and fetal outcomes.
METHODS: This was a cross-sectional study consisted of 21 pregnant women with hypertension and 23 without hypertension. The gestational age ranged from 28 to 39 weeks (hypertensive) and 32 to 40 weeks (normotensive). The paraffin embedded formalin fixed placenta tissue blocks were retrieved from the pathology archives. Endocan immunohistochemistry was performed on tissue sections of full thickness and maternal surface of the placenta. The endocan expression was determined in fetal endothelial cells, maternal endothelial cells, cytotrophoblasts, syncytiotrophoblasts and decidual cells. The differences in endocan expression in placenta between hypertensive and normotensive subjects were evaluated by Pearson chi-square test and t-test were used in the statistical analysis.
RESULTS: The endocan expression was significantly higher in fetal endothelial cells (P

Matched MeSH terms: Paraffin Embedding
Effects of Crude Palm Oil and Crude Palm Kernel Oil Upon Wax Inhibition

Ragunathan T, Husin H, Wood CD

ACS Omega, 2020 Aug 11;5(31):19342-19349.
PMID: 32803027 DOI: 10.1021/acsomega.0c00753

The ever-increasing demand for the finite source of oil has led oil production companies to produce and transport the produced crude oil as efficiently and economically as possible. One of the major concerns especially in waters like the South China Sea is the deposition of wax on the walls of the pipeline or wellbore, constricting and hindering the hydrocarbon flow. This is due to the low seabed temperatures, which can be below the wax appearance temperature (WAT), leading to the deposition of wax out of waxy crude oil through the molecular dispersion mechanism. Currently, many prevention and remedy methods are in place to overcome the problem, but most of the additives possess environmental threat, as most of the chemical solutions used are toxic, nonorganic, and costly. Hence, this paper aims to provide some insights into the effect of palm oil derivatives such as crude palm oil (CPO) and crude palm kernel oil (CPKO) on wax inhibition. The effect of aging time (i.e., immersion time) was also evaluated. A comparison was made between paraffin inhibition efficiency results (PIE %) obtained by CPO, CPKO, poly(ethylene-co-vinyl acetate) (EVA), and triethanolamine (TEA). It was observed that the average efficiency of 81.67% was obtained when 1% CPO was added to heavy crude oil. The wax inhibition performance reached a plateau after 1.5 h of aging time for all of the investigated samples.

Matched MeSH terms: Paraffin
Fulltext Effect of the miR-96-5p inhibitor and mimic on the migration and invasion of the SW480-7 colorectal cancer cell line

He PY, Yip WK, Jabar MF, Mohtarrudin N, Dusa NM, Seow HF

Oncol Lett, 2019 Aug;18(2):1949-1960.
PMID: 31423265 DOI: 10.3892/ol.2019.10492

The objectives of the present study were to identify the aberrant expression of microRNA (miRNA) in colorectal carcinoma (CRC) tissues from published miRNA profiling studies and to investigate the effects of the identified miRNA inhibitor and mimic miR-96-5p on CRC cell migration and invasion. The altered expression of the regulators of cytoskeleton mRNA in miR-96-5p inhibitor-transfected cells was determined. The miR-96-5p expression level in five CRC cell lines, HCT11, CaCo2, HT29, SW480 and SW620, and 26 archived paraffin-embedded CRC tissues were also investigated by reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR). Cell viability in response to the miR-96-5p inhibitor and mimic transfections was determined by an MTT assay. A Matrigel invasion assay was conducted to select the invasive subpopulation designated SW480-7, by using the parental cell line SW480. The effects of miR-96-5p mimic- or inhibitor-transfected SW480-7 cells on cell migration and invasion were evaluated using the Transwell and Matrigel assays, and the change in expression of the regulators of cytoskeleton mRNAs was identified by Cytoskeleton Regulators RT2-Profiler PCR array followed by validation with RT-qPCR. CRC tissues exhibited a significant increase in miR-96-5p expression, compared with their matched normal adjacent tissues, indicating an oncogenic role for miR-96-5p. The results demonstrated that the miR-96-5p inhibitor decreased the migration of SW480-7 cells, but had no effect on invasion. This may be due to the promotion of cell invasion by Matrigel, which counteracts the blockade of cell invasion by the miR-96-5p inhibitor. The miR-96-5p mimic enhanced SW480-7 cell migration and invasion, as expected. It was determined that there was a >2.5 fold increase in the expression of genes involved in cytoskeleton regulation, myosin light chain kinase 2, pleckstrin homology like domain family B member 2, cyclin A1, IQ motif containing GTPase activating protein 2, Brain-specific angiogenesisinhibitor 1-associated protein 2 and microtubule-actin crosslinking factor 1, in miR-96-5p inhibitor-transfected cells, indicating that they are negative regulators of cell migration. In conclusion, the miR-96-5p inhibitor blocked cell migration but not invasion, and the latter may be due to the counteraction of Matrigel, which has been demonstrated to stimulate cell invasion.

Matched MeSH terms: Paraffin Embedding
Effect of mixing process parameters and suitability of backbone polymer for aluminum powder injection molding feedstock

Abdullahi A, Choudhury I, Azuddin M, Nahar N

Sains Malaysiana, 2017;46:477-483.

A suitable and cost-effective microfabrication technique for processing aluminum micropart is required, as the choice
of aluminum microparts for aerospace, electronics and automobile components is preferred over other metals due to its
excellent properties. Meanwhile, powder injection molding (PIM) is identified as an economical manufacturing technique
for processing ceramic and micro-metal powders into microparts and or components. Therefore, this study investigates
formulation and processing of aluminum PIM feedstock using a custom-made machine. The investigation is focused on
the effect of mixing process parameters (powder loading, rotor speed and mixing temperature) and the suitability of
the backbone polymer. The formulated PIM feedstock constituents are paraffin wax (PW), stearic acid (SA), high-density
polyethylene (HDPE)/ medium-density polyethylene (MDPE) alternatively and aluminum micro-metal powder. Taguchi
method is used for the design of experiments (DOEs) and analysis. In addition, response surface methodology (RSM) is
employed to develop empirical viscosity models. The optimum powder-binder mixing ratio of 58:42 vol. % with rotor
speed of 43 rpm were determined for preparing aluminum PIM feedstock using mini-lab mixer developed. The empirical
model developed for aluminum PIM feedstock viscosity shows a good fit with R2
values of 0.84 using HDPE and 0.96 for
MDPE binder system. This investigation demonstrates preparation and suitability of aluminum PIM feedstock using waxbased
binder system.

Matched MeSH terms: Paraffin
ERG oncoprotein expression in prostate carcinoma patients of different ethnicities

Kelly GM, Kong YH, Dobi A, Srivastava S, Sesterhenn IA, Pathmanathan R, et al.

Molecular and clinical oncology, 2015 Jan;3(1):23-30.
PMID: 25469265

Overexpression of the erythroblast transformation-specific-related gene (ERG) oncoprotein due to transmembrane protease, serine 2 (TMPRSS2)-ERG fusion, the most prevalent genomic alteration in prostate cancer (CaP), is more frequently observed among Caucasian patients compared to patients of African or Asian descent. To the best of our knowledge, this is the first study to investigate the prevalence of ERG alterations in a multiethnic cohort of CaP patients. A total of 191 formalin-fixed paraffin-embedded sections of transrectal ultrasound-guided prostate biopsy specimens, collected from 120 patients treated at the Sime Darby Medical Centre, Subang Jaya, Malaysia, were analyzed for ERG protein expression by immunohistochemistry using the anti-ERG monoclonal antibody 9FY as a surrogate for the detection of ERG fusion events. The overall frequency of ERG protein expression in the population evaluated in this study was 39.2%. Although seemingly similar to rates reported in other Asian communities, the expression of ERG was distinct amongst different ethnic groups (P=0.004). Malaysian Indian (MI) patients exhibited exceedingly high expression of ERG in their tumors, almost doubling that of Malaysian Chinese (MC) patients, whereas ERG expression was very low amongst Malay patients (12.5%). When collectively analyzing data, we observed a significant correlation between younger patients and higher ERG expression (P=0.04). The prevalence of ERG expression was significantly different amongst CaP patients of different ethnicities. The higher number of ERG-expressing tumors among MI patients suggested that the TMPRSS2-ERG fusion may be particularly important in the pathogenesis of CaP amongst this group of patients. Furthermore, the more frequent expression of ERG among the younger patients analyzed suggested an involvement of ERG in the early onset of CaP. The results of this study underline the value of using ERG status to better understand the differences in the etiology of CaP initiation and progression between ethnic groups.

Matched MeSH terms: Paraffin Embedding
Dual variant of Epstein-Barr virus in Hodgkin/Reed-Sternberg cells: single-cell PCR study on latent membrane protein-1 gene

Kim LH, Peh SC, Poppema S

Int J Cancer, 2003 Nov 1;107(2):250-5.
PMID: 12949802

Isolation of single cells permits analysis of DNA or RNA from individual cells among heterogeneous populations. This technique is particularly useful in the study of classical Hodgkin's lymphoma (cHL) due to the scarcity of H/RS tumor cells among large numbers of reactive leukocytes. In a previous study, we found a high frequency of dual LMP-1 variant (concurrent presence of deleted and nondeleted variants) in cHL from whole-tissue sections. For the present study, we applied a single-cell isolation technique to determine the LMP-1 oncogene variant in EBV-associated H/RS cells. Five cases of EBV-infected cHL, containing nondeleted (n=1), deleted (n=1) and dual infection (n=3) based on whole-tissue section analysis, were selected for study. Paraffin-embedded tissue sections were stained with antibody to LMP-1 and positively stained H/RS cells isolated using a semiautomated micromanipulator. Each isolated single cell was subjected to PCR for amplification of the LMP-1 gene flanking the 30 bp deletion region and Xho1 restriction site. Cases with either nondeleted variant or the deleted variant showed similar LMP-1 variant expression in isolated single H/RS cells. However, 1 of the 3 cases with dual variants showed only the deleted variant in H/RS cells. The other 2 cases showed mixed patterns of deleted, nondeleted and dual LMP-1 variants in isolated single H/RS cells. All cases showed loss of the Xho1 restriction site, with the exception of the case with nondeleted LMP-1. Results of single-H/RS cell analysis of the Xho1 restriction site concur with those of whole-tissue section amplification. A mixed pattern of LMP-1 variants was observed in isolated H/RS cells, and it is speculated that this is due to the accumulation of mutation and deletion events.

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