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Improved method for the isolation of RNA from bacteria refractory to disruption, including S. aureus producing biofilm

Atshan SS, Shamsudin MN, Lung LT, Ling KH, Sekawi Z, Pei CP, et al.

Gene, 2012 Feb 25;494(2):219-24.
PMID: 22222139 DOI: 10.1016/j.gene.2011.12.010

The development of fast, reliable and inexpensive phenol protocol is described for the isolation of RNA from bacterial biofilm producers. The method was tested on Staphylococcus aureus (S. aureus) and other biofilm-producing gram-negative microorganisms and provided the highest integrity of RNA recovery in comparison to other methods reported here. In parallel experiments, bacterial lysis with Qiagen, NucleoSpin RNAII, InnuREP RNA Mini, Trizol and MasterPure RNA extraction Kits using standard protocols consistently gave low RNA yields with an absence of integrity. The boiling method presented here yielded high concentration of RNA that was free from 16S and 23S rRNA, contained 5S RNA. Higher yields due to improved biofilm bacterial cell lysis were achieved with an added hot phenol incubation step without the need for a bead mill or the enzyme. This method when used in conjunction with the Qiagen RNeasy Mini kit, RNA isolation was a success with greater integrity and contained undegraded 16S and 23S rRNA and did not require further purification. Contaminating DNA was a problem with the RNA processing samples; we used quantitative real-time PCR (RT-qPCR) to measure the recovery of RNA from bacterial biofilm cells using the method described here.

Matched MeSH terms: Staphylococcus aureus/genetics*
Phylogenetic diversity of acidophilic actinomycetes from Malaysia

Muramatsu H, Murakami R, Ibrahim ZH, Murakami K, Shahab N, Nagai K

J Antibiot (Tokyo), 2011 Sep;64(9):621-4.
PMID: 21792208 DOI: 10.1038/ja.2011.57
Matched MeSH terms: Actinobacteria/genetics*
Antioxidative peptides from food proteins: a review

Sarmadi BH, Ismail A

Peptides, 2010 Oct;31(10):1949-56.
PMID: 20600423 DOI: 10.1016/j.peptides.2010.06.020

Bioactive peptides, as products of hydrolysis of diverse food proteins, are the focus of current research. They exert various biological roles, one of the most crucial of which is the antioxidant activity. Reverse relationship between antioxidant intake and diseases has been approved through plenty of studies. Antioxidant activity of bioactive peptides can be attributed to their radical scavenging, inhibition of lipid peroxidation and metal ion chelation properties of peptides. It also has been proposed that peptide structure and its amino acid sequence can affect its antioxidative properties. This paper reviews bioactive peptides from food sources concerning their antioxidant activities. Additionally, specific characteristics of antioxidative bioactive peptides, enzymatic production, methods to evaluate antioxidant capacity, bioavailability, and safety concerns of peptides are reviewed.

Matched MeSH terms: Peptides/genetics
Taxonomic status of three types of Fejervarya cancrivora from Indonesia and other Asian countries based on morphological observations and crossing experiments

Kurniawan N, Djong TH, Islam MM, Nishizawa T, Belabut DM, Sen YH, et al.

Zoolog Sci, 2011 Jan;28(1):12-24.
PMID: 21186942 DOI: 10.2108/zsj.28.12

Although the crab-eating frog Fejervarya cancrivora is one of the most widely distributed species in Asian region, taxonomic relationships among different populations remain unclarified. In this study, we attempted to elucidate the taxonomic status of F. cancrivora from Indonesian and other Asian populations. Five populations of F. cancrivora from Selangor (Malaysia), Cianjur (Java, Indonesia), Trat (Thailand), Khulna (Bangladesh), and Makassar (Sulawesi, Indonesia) were morphologically observed and subjected to crossing experiments. Principal component and clustering analyses revealed that these five populations could be organized into three groups corresponding to three observed morphological types: a Selangor and Cianjur group (large-type), a Trat and Khulna group (mangrove-type), and a Makassar group (Sulawesi-type). The limited crossing experiments revealed that hybrids between Selangor females and Cianjur and Trat males developed normally, whereas hybrids between Selangor females and Khulna males showed incomplete gametic isolation. Histological observations of the testes of mature males revealed the presence of pycnotic nuclei in the hybrids between Selangor females and Khulna males in addition to normal bundles of spermatozoa. In contrast, no pycnotic nuclei were observed in the Selangor controls. Although meiotic metaphases in the controls were normal, those in hybrids showed several abnormalities, such as the appearance of univalents and an increase in rod-shaped bivalents. Based on our findings from the morphological observations and crossing experiments, we conclude that each of three identified types represents a distinct species. We propose that the large-type is F. cancrivora, the mangrove-type is F. moodiei, and the Sulawesi-type represents an undescribed species.

Matched MeSH terms: Ranidae/genetics
Fulltext Improving the potency of DNA vaccine against chicken anemia virus (CAV) by fusing VP1 protein of CAV to Marek's Disease Virus (MDV) type-1 VP22 protein

Moeini H, Omar AR, Rahim RA, Yusoff K

Virol J, 2011;8:119.
PMID: 21401953 DOI: 10.1186/1743-422X-8-119

Studies have shown that the VP22 gene of Marek's Disease Virus type-1 (MDV-1) has the property of movement between cells from the original cell of expression into the neighboring cells. The ability to facilitate the spreading of the linked proteins was used to improve the potency of the constructed DNA vaccines against chicken anemia virus (CAV).

Matched MeSH terms: Herpesvirus 2, Gallid/genetics; Recombinant Fusion Proteins/genetics; Viral Proteins/genetics; Viral Vaccines/genetics; Chicken anemia virus/genetics; Vaccines, DNA/genetics; Capsid Proteins/genetics
Gene expression profiling and cancer-related pathways in type I endometrial carcinoma

Saghir FS, Rose IM, Dali AZ, Shamsuddin Z, Jamal AR, Mokhtar NM

Int. J. Gynecol. Cancer, 2010 Jul;20(5):724-31.
PMID: 20973258

INTRODUCTION: Malignant transformation of type I endometrium involves alteration in gene expression with subsequent uncontrolled proliferation of altered cells.
OBJECTIVE: The main objective of the present study was to identify the cancer-related genes and gene pathways in the endometrium of healthy and cancer patients.
MATERIALS AND METHODS: Thirty endometrial tissues from healthy and type I EC patients were subjected to total RNA isolation. The RNA samples with good integrity number were hybridized to a new version of Affymetrix Human Genome GeneChip 1.0 ST array. We analyzed the results using the GeneSpring 9.0 GX and the Pathway Studio 6.1 software. For validation assay, quantitative real-time polymerase chain reaction was used to analyze 4 selected genes in normal and EC tissue.
RESULTS: Of the 28,869 genes profiled, we identified 621 differentially expressed genes (2-fold) in the normal tissue and the tumor. Among these genes, 146 were up-regulated and 476 were down-regulated in the tumor as compared with the normal tissue (P < 0.001). Up-regulated genes included the v-erb-a erythroblastic leukemia viral oncogene homolog 3 (ErbB3), ErbB4, E74-like factor 3 (ELF3), and chemokine ligand 17 (CXCL17). The down-regulated genes included signal transducer and activator transcription 5B (STAT5b), transforming growth factor A receptor III (TGFA3), caveolin 1 (CAV1), and protein kinase C alpha (PKCA). The gene set enrichment analysis showed 10 significant gene sets with related genes (P < 0.05). The quantitative polymerase chain reaction of 4 selected genes using similar RNA confirmed the microarray results (P < 0.05).
CONCLUSIONS: Identification of molecular pathways with their genes related to type I EC contribute to the understanding of pathophysiology of this cancer, probably leading to identifying potential biomarkers of the cancer.

Matched MeSH terms: Endometrial Neoplasms/genetics*
Initiating a Human Variome Project Country Node

AlAama J, Smith TD, Lo A, Howard H, Kline AA, Lange M, et al.

Hum Mutat, 2011 May;32(5):501-6.
PMID: 21305654 DOI: 10.1002/humu.21463

Genetic diseases are a pressing global health problem that requires comprehensive access to basic clinical and genetic data to counter. The creation of regional and international databases that can be easily accessed by clinicians and diagnostic labs will greatly improve our ability to accurately diagnose and treat patients with genetic disorders. The Human Variome Project is currently working in conjunction with human genetics societies to achieve this by establishing systems to collect every mutation reported by a diagnostic laboratory, clinic, or research laboratory in a country and store these within a national repository, or HVP Country Node. Nodes have already been initiated in Australia, Belgium, China, Egypt, Malaysia, and Kuwait. Each is examining how to systematically collect and share genetic, clinical, and biochemical information in a country-specific manner that is sensitive to local ethical and cultural issues. This article gathers cases of genetic data collection within countries and takes recommendations from the global community to develop a procedure for countries wishing to establish their own collection system as part of the Human Variome Project. We hope this may lead to standard practices to facilitate global collection of data and allow efficient use in clinical practice, research and therapy.

Matched MeSH terms: Genome, Human/genetics*
Fulltext Progressive familial intrahepatic cholestasis in Malaysian patients--a report of five cases

Lee WS, Chai PF, Looi LM

Med J Malaysia, 2009 Sep;64(3):216-9.
PMID: 20527271

Progressive familial intrahepatic cholestasis (PFIC) is characterized by early onset cholestasis, progressive liver cirrhosis, pruritus, poor growth and inexorable progression to liver cirrhosis in early childhood. The serum level of gamma-glutamyl transferase is low or normal, which is discordant with severe cholestasis. Five Malaysian patients with PFIC, who all had typical features of PFIC with early onset of severe and progressive cholestasis, pruritus, cirrhosis and liver failure, were described. Three patients died as a result of the disease, while another one died due to post-liver transplant complication. The only survivor has compensated liver cirrhosis. Patients with severe cholestasis but has spuriously low yGT should be suspected of having PFIC. Liver transplant, which is life-saving in a majority of patients with PFIC, should be considered in all patients with PFIC.

Matched MeSH terms: Cholestasis, Intrahepatic/genetics*
Flowering response and crop duration of aromatic rices in diverse environments

Shahidullah SM, Hanafi MM, Ashrafuzzaman M, Salam MA, Khair A

C. R. Biol., 2009 Oct;332(10):909-16.
PMID: 19819411 DOI: 10.1016/j.crvi.2009.07.003

Crop duration of a rice plant, essentially dictated by flowering response, is an important selection criterion. It is determined by the interaction of genotype and environment. A field experiment was conducted with 40 rice genotypes to assess the fluctuation and/or stability of crop duration in a series of 16 environmental conditions. The effects of genotype, environment and all the components of G x E interaction were highly significant. Among the genotypes Benaful and Gandho kasturi were most sensitive to environmental changes, and indicating lower adaptability over the environments. Crop durations of 17 genotypes were comparatively stable against environmental changes. Four genotypes viz. Basmati PNR346, BR28, Neimat and Sarwati showed only nonlinear sensitivity and thus unpredictable fluctuation. Seventeen genotypes indicated average stability over the environments. The AMMI analysis identified Badshabhog, Basmati Tapl-90, Bhog ganjia, BR38, Elai, Jata katari and Radhuni pagal as most stable genotypes over the environment series. It also advocated three comparatively stable environments for all the genotypes.

Matched MeSH terms: Oryza/genetics
Biomass accumulation and energy conversion efficiency in aromatic rice genotypes

Shahidullah SM, Hanafi MM, Ashrafuzzaman M, Razi Ismail M, Salam MA, Khair A

C. R. Biol., 2010 Jan;333(1):61-7.
PMID: 20176338 DOI: 10.1016/j.crvi.2009.10.002

A field experiment was conducted to evaluate photosynthetic efficiency along with different growth parameters of aromatic rice genotypes. Forty genotypes including three non-aromatic checks exhibited enormous variations for leaf area index (LAI), crop growth rate (CGR), relative growth rate (RGR), net assimilation rate (NAR), grain yield, total dry matter, harvest index and photosynthetic efficiency or energy use efficiency (Emu) at panicle initiation and heading stages. Minimum LAI-value was 0.52 in Khazar at PI stage and maximum was 4.91 in Sakkor khora at heading stage. The CGR-value was in the range of 4.80-24.11 g m(-2) per day. The best yielder BR39 produced grain of 4.21 t ha(-1) and the worst yielder Khazar gave 1.42 t ha(-1). Total dry matter (TDM) yield varied from 4.04 to 12.26 t ha(-1) where genotypes proved their energy use efficiency a range between 0.58 to 1.65%. Emu showed a significant positive relation with TDM (r=0.80(**)), CGR (r=0.72(**)) and grain yield (r=0.66(**)). A negative correlation was established between TDM and harvest index and LAI and RGR. Path analysis result showed that NAR at heading stage exerted highest positive direct effect (0.70) on Emu.

Matched MeSH terms: Oryza/genetics*
Fulltext Iris mammillations in two female siblings with congenital adrenal hyperplasia

Peyman M, Ong MJ, Iqbal T, Subrayan V

BMJ Case Rep, 2010;2010.
PMID: 22802477 DOI: 10.1136/bcr.08.2010.3266

Iris mammillations are dark brown, smooth, mound- or dome-shaped protuberances that are typically found on the anterior iris surface and are presumed to be congenital in origin. This congenital anomaly is usually unilateral and can be hereditary or sporadic. Lisch nodules in neurofibromatosis, tapioca melanoma of the iris, inflammatory iris granulomata and Cogan-Reese syndrome should be considered in the differential diagnosis. In this case report, the authors present a case of a bilateral iris mammillations in two siblings with congenital adrenal hyperplasia (CAH). To our knowledge, this is the first case where bilateral iris mammilations have been found to be associated with a systemic condition. Iris mammillations can be considered as one of the clinical signs in CAH in view of the pathogenesis discussed. Detailed ocular examination in CAH may reveal an increased incidence.

Matched MeSH terms: Adrenal Hyperplasia, Congenital/genetics*
Fulltext Construction of a polycystic ovarian syndrome (PCOS) pathway based on the interactions of PCOS-related proteins retrieved from bibliomic data

Mohamed-Hussein ZA, Harun S

Theor Biol Med Model, 2009;6:18.
PMID: 19723303 DOI: 10.1186/1742-4682-6-18

Polycystic ovary syndrome (PCOS) is a complex but frequently occurring endocrine abnormality. PCOS has become one of the leading causes of oligo-ovulatory infertility among premenopausal women. The definition of PCOS remains unclear because of the heterogeneity of this abnormality, but it is associated with insulin resistance, hyperandrogenism, obesity and dyslipidaemia. The main purpose of this study was to identify possible candidate genes involved in PCOS. Several genomic approaches, including linkage analysis and microarray analysis, have been used to look for candidate PCOS genes. To obtain a clearer view of the mechanism of PCOS, we have compiled data from microarray analyses. An extensive literature search identified seven published microarray analyses that utilized PCOS samples. These were published between the year of 2003 and 2007 and included analyses of ovary tissues as well as whole ovaries and theca cells. Although somewhat different methods were used, all the studies employed cDNA microarrays to compare the gene expression patterns of PCOS patients with those of healthy controls. These analyses identified more than a thousand genes whose expression was altered in PCOS patients. Most of the genes were found to be involved in gene and protein expression, cell signaling and metabolism. We have classified all of the 1081 identified genes as coding for either known or unknown proteins. Cytoscape 2.6.1 was used to build a network of protein and then to analyze it. This protein network consists of 504 protein nodes and 1408 interactions among those proteins. One hypothetical protein in the PCOS network was postulated to be involved in the cell cycle. BiNGO was used to identify the three main ontologies in the protein network: molecular functions, biological processes and cellular components. This gene ontology analysis identified a number of ontologies and genes likely to be involved in the complex mechanism of PCOS. These include the insulin receptor signaling pathway, steroid biosynthesis, and the regulation of gonadotropin secretion among others.

Matched MeSH terms: Polycystic Ovary Syndrome/genetics
Circadian and solar clocks interact in seasonal flowering

Yeang HY

Bioessays, 2009 Nov;31(11):1211-8.
PMID: 19795408 DOI: 10.1002/bies.200900078

The plant maintains a 24-h circadian cycle that controls the sequential activation of many physiological and developmental functions. There is empirical evidence suggesting that two types of circadian rhythms exist. Some plant rhythms appear to be set by the light transition at dawn, and are calibrated to circadian (zeitgeber) time, which is measured from sunrise. Other rhythms are set by both dawn and dusk, and are calibrated to solar time that is measured from mid-day. Rhythms on circadian timing shift seasonally in tandem with the timing of dawn that occurs earlier in summer and later in winter. On the other hand, rhythms set to solar time are maintained independently of the season, the timing of noon being constant year-round. Various rhythms that run in-phase and out-of-phase with one another seasonally may provide a means to time and induce seasonal events such as flowering.

Matched MeSH terms: Arabidopsis/genetics
Prevalence and quantification of Vibrio parahaemolyticus in raw salad vegetables at retail level

Tunung R, Margaret S, Jeyaletchumi P, Chai LC, Tuan Zainazor TC, Ghazali FM, et al.

J Microbiol Biotechnol, 2010 Feb;20(2):391-6.
PMID: 20208446

The purpose of this study was to investigate the biosafety of Vibrio parahaemolyticus in raw salad vegetables at wet market and supermarket in Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was applied to detect the presence of V. parahaemolyticus and to enumerate their density in the food samples. The study analyzed 276 samples of common vegetables eaten raw in Malaysia (Wild cosmos = 8; Japanese parsley = 21; Cabbage = 30; Lettuce = 16; Indian pennywort = 17; Carrot = 31; Sweet potato = 29; Tomato = 38; Cucumber = 28; Four winged bean = 26; Long bean = 32). The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). The occurrence of V. parahaemolyticus detected was 20.65%, with higher frequency of V. parahaemolyticus in vegetables obtained from wet markets (Wet market C = 27.27%Wet Market D = 32.05%) compared to supermarkets (Supermarket A = 1.64%; Supermarket B = 16.67%). V. parahaemolyticus was most prevalent in Indian pennywort (41.18%). The density of V. parahaemolyticus in all the samples ranged from <3 up to >2400 MPN/g, mostly <3 MPN/g concentration. Raw vegetables from wet markets contained higher levels of V. parahaemolyticus compared to supermarkets. V. parahaemolyticus were present in raw vegetables although in low numbers. The results suggest that raw vegetables act as a transmission route for V. parahaemolyticus. This study will be the first biosafety assessment of V. parahaemolyticus in raw vegetables in Malaysia.

Matched MeSH terms: Vibrio parahaemolyticus/genetics
Rapid and effective method of RNA isolation from green microalga Ankistrodesmus convolutus

Thanh T, Omar H, Abdullah MP, Chi VT, Noroozi M, Ky H, et al.

Mol Biotechnol, 2009 Oct;43(2):148-53.
PMID: 19507070 DOI: 10.1007/s12033-009-9182-8

The rapid and effective method for the isolation of RNA from green microalga Ankistrodesmus convolutus based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride is developed. This procedure avoids the use of toxic chaotropic agents and phenol while high concentration of dithiothreitol is used to inhibit RNase activity and prevent oxidative cross-linking of nucleic acids by phenolics. The extraction procedure was able to produce high quality and intact RNA from A. convolutus. The yield of total RNA was 0.69-0.73 mg/g of fresh weight, with A(260)/A(280) ratio of 1.79-1.86. The obtained RNA was of sufficient quality and suitable for downstream application such as RT-PCR and cDNA library construction. The procedure may also have wider applicability for total RNA isolation from other green microalgae species.

Matched MeSH terms: Chlorophyta/genetics*
Re-emergence of Chikungunya virus in South-east Asia: virological evidence from Sri Lanka and Singapore

Hapuarachchi HC, Bandara KB, Sumanadasa SD, Hapugoda MD, Lai YL, Lee KS, et al.

J Gen Virol, 2010 Apr;91(Pt 4):1067-76.
PMID: 19955565 DOI: 10.1099/vir.0.015743-0

Chikungunya fever swept across many South and South-east Asian countries, following extensive outbreaks in the Indian Ocean Islands in 2005. However, molecular epidemiological data to explain the recent spread and evolution of Chikungunya virus (CHIKV) in the Asian region are still limited. This study describes the genetic Characteristics and evolutionary relationships of CHIKV strains that emerged in Sri Lanka and Singapore during 2006-2008. The viruses isolated in Singapore also included those imported from the Maldives (n=1), India (n=2) and Malaysia (n=31). All analysed strains belonged to the East, Central and South African (ECSA) lineage and were evolutionarily more related to Indian than to Indian Ocean Islands strains. Unique genetic characteristics revealed five genetically distinct subpopulations of CHIKV in Sri Lanka and Singapore, which were likely to have emerged through multiple, independent introductions. The evolutionary network based on E1 gene sequences indicated the acquisition of an alanine to valine 226 substitution (E1-A226V) by virus strains of the Indian sublineage as a key evolutionary event that contributed to the transmission and spatial distribution of CHIKV in the region. The E1-A226V substitution was found in 95.7 % (133/139) of analysed isolates in 2008, highlighting the widespread establishment of mutated CHIKV strains in Sri Lanka, Singapore and Malaysia. As the E1-A226V substitution is known to enhance the transmissibility of CHIKV by Aedes albopictus mosquitoes, this observation has important implications for the design of vector control strategies to fight the virus in regions at risk of chikungunya fever.

Matched MeSH terms: Chikungunya virus/genetics
Clinical manifestations in trisomy 9

Kannan TP, Hemlatha S, Ankathil R, Zilfalil BA

Indian J Pediatr, 2009 Jul;76(7):745-6.
PMID: 19475342 DOI: 10.1007/s12098-009-0158-2

Complete trisomy 9 is a lethal diagnosis and most fetuses diagnosed thus die prenatally or during the early postnatal period and majority of such cases have been known to end in spontaneous abortion in the first trimester itself. One such rare survival of fetus ending in normal delivery and surviving until 20 days is reported here detailing the clinical manifestations of the child during the period of survival. The salient clinical features observed were small face, wide fontanel, prominent occiput, micrognathia, low set ears, upslanting palpebral fissures, high arched palate, short sternum, overlapping fingers, limited hip abduction, rocker bottom feet, heart murmurs and also webbed neck, characteristic of this trisomy 9 syndrome.

Matched MeSH terms: Abnormalities, Multiple/genetics*
Fulltext Chikungunya virus of Central/East African genotype detected in Malaysia

Soon YY, Junaidi I, Kumarasamy V, Chem YK, Juliana R, Chua KB

Med J Malaysia, 2007 Aug;62(3):214-7.
PMID: 18246910 MyJurnal

Since its isolation in Tanzania in 1953, chikungunya virus has caused periodic outbreaks in both tropical Africa and Asia. In the last decade, the virus has shown not only increased activity but has expanded its geographical locations, thus classical delineation of various genotypes of chikungunya virus to specific geographic locales no longer holds true. Rapid mass movement of people and the constant presence of the right vectors in this region could have contributed to the change in virus ecology. This paper documents the first detection of chikungunya virus of Central/East genotype in Malaysia from a patient who was most likely infected with the virus during her visit to India. Without good Aedes vector measures, only time will tell whether this genotype rather than the existing endemic genotype will subsequently cause the next chikungunya outbreak in Malaysia.

Matched MeSH terms: Chikungunya virus/genetics*
Detection of pbp2b and ermB genes in clinical isolates of Streptococcus pneumoniae

Kumari N, Navaratnam P, Sekaran SD

J Infect Dev Ctries, 2008 Jun 01;2(3):193-9.
PMID: 19738350

BACKGROUND: Streptococcus pneumoniae is a major human pathogen. The emergence of penicillin resistant strains since the 1970s has been life threatening and the evolution of the bacteria have enabled itself to develop resistance to many other antibiotics such as the macrolides and the fluoroquinolones. This study aims to characterize S. pneumoniae isolates for the presence of penicillin and macrolide resistance genes.
METHODOLOGY: One hundred and twenty clinical isolates of S. pneumoniae were obtained from patients of University Malaya Medical Centre (UMMC). The strains were screened using a multiplex real-time PCR method for the presence of alterations in the genes encoding the penicillin binding proteins: pbp2b, macrolide resistance determinant ermB and the pneumolysin gene, ply. Dual-labelled Taqman probes were used in the real-time detection method comprising three different genes labeled with individual fluorophores at different wavelengths. One hundred and twenty isolates from bacterial cultures and isolates directly from blood cultures samples were analyzed using this assay.
RESULTS: A multiplex PCR comprising the antibiotic resistance genes, ermB and and pneumolysin gene (ply), a S. pneumoniae species specific gene, was developed to characterize strains of S. pneumoniae. Out of the 120 pneumococcal isolates, 58 strains were categorized as Penicillin Sensitive Streptococcus pneumoniae (PSSP), 36 as Penicillin Intermediate Streptococcus pneumoniae (PISP) and 26 as Penicillin Resistant Streptococcus pneumoniae (PRSP). All the 58 PSSP strains harboured the pbp2b gene while the 36 PISP and 26 PRSP strains did not harbour this gene, thus suggesting reduced susceptibility to penicillin. Resistance to erythromycin was observed in 47 of the pneumococcal strains while 15 and 58 were intermediate and sensitive to this drug respectively. Susceptibility testing to other beta-lactams (CTX and CRO) also showed reduced susceptibility among the strains within the PISP and PRSP groups but most PSSP strains were sensitive to other antibiotics.
CONCLUSION: The characterization of pneumococcal isolates for penicillin and erythromycin resistance genes could be useful to predict the susceptibility of these isolates to other antibiotics, especially beta-lactams drugs. We have developed an assay with a shorter turnaround time to determine the species and resistance profile of Streptococcus pneumoniae with respect to penicillin and macrolides using the Real Time PCR format with fluorescent labeled Taqman probes, hence facilitating earlier and more definitive antimicrobial therapy which may lead to better patient management.

Matched MeSH terms: Bacterial Proteins/genetics; DNA, Bacterial/genetics; Streptococcus pneumoniae/genetics*; Streptolysins/genetics; Aminoacyltransferases/genetics*; Drug Resistance, Multiple, Bacterial/genetics; Penicillin-Binding Proteins/genetics*
Human nasal turbinates as a viable source of respiratory epithelial cells using co-culture system versus dispase-dissociation technique

Noruddin NA, Saim AB, Chua KH, Idrus R

Laryngoscope, 2007 Dec;117(12):2139-45.
PMID: 17891046

OBJECTIVE: To compare a co-culture system with a conventional dispase-dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application.
METHODS: Human respiratory epithelial cells were serially passaged using a co-culture system and a conventional dispase-dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin-18, a marker for ciliated and secretory epithelial cells; cytokeratin-14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular-weight cytokeratin 34 beta E12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells.
RESULTS: Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co-cultured respiratory epithelial showed a 2.6-times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co-cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques.
CONCLUSION: Co-culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.

Matched MeSH terms: DNA/genetics; Mucins/genetics; Nuclear Proteins/genetics; Intracellular Signaling Peptides and Proteins/genetics; Cell Growth Processes/genetics; Keratin-18/genetics; Keratin-14/genetics

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