Nasopharynx as a Microbiologic Reservoir in Chronic Suppurative Otitis Media: Preliminary Study

ObjectivesThe present study was designed to identify the correlations of bacterial strains of the middle ear and the nasopharynx in chronic suppurative otitis media (CSOM) patients who were scheduled for operations.MethodsSixty-three patients with CSOM were enrolled in the study. Culture specimens were collected from the middle ear and nasopharynx of patients who were admitted for operation. Samples collections were performed 3 times; from the middle ear and nasophaynx at the admission day, from the middle ear during the operation, and from the external auditory canal post-operatively. Bacteria were identified by gram staining and biochemical tests. The correspondence rate of organisms which simultaneously exist in the middle ear and the nasopharynx was measured.ResultsSixty-eight organisms were isolated from the middle ear and 57 organisms from the nasopharynx among 63 patients. Of 68 bacteria identified in middle ear, 26.52% (18 bacteria) corresponded with those of nasopharynx. MRSA had the high correspondence rate, and of 18 methicillin-resistant Staphylococcus aureus (MRSA) isolated from middle ear, 33.3% (6 bacteria) corresponded with nasophaynx. Meanwhile, 3 organisms of MRSA were detected from the external auditory canal post-operatively, although they were only found in nasopharynx pre-operatively.ConclusionThe current trend of middle ear swab alone for bacterial detection would be insufficient to identify the potent MRSA and impede early antibiotic intervention for the effective middle ear surgery. Therefore, it is necessary to perform nasopharynx cultures together with conventional middle ear culture to control potent risk for infection pre-operatively

Cloning, Expression, Purification, and Characterization of Clostridium botulinum Neurotoxin Serotype F Domains

The use of recombinant BoNT domains has been proposed as a means to develop strategies to treat and prevent botulism. Here, details on the molecular cloning, protein expression, purification, and immunoreactivity of BoNT/F domains from Clostridium botulinum are presented. Initially, full-length synthetic genes encoding recombinant BoNT/F domains (catalytic, translocation, and receptor binding) were designed and cloned into Escherichia coli for expression. Recombinant proteins were then purified through GST affinity chromatography preceding elution of GST-free recombinant domains by thrombin protease. Soluble recombinant proteins encoding catalytic light chain and translocation N-terminal heavy chain were subsequently used to perform in vivo immunization. Polyclonal mouse antibodies specific to these domains were raised, confirmed by Western blot analysis and elevated immunoreactivity was identified through indirect ELISA. In conclusion, availability of the recombinant protein provides an effective system to study the immunological aspects of BoNT/F and corresponding applications in pathogen detection and vaccine candidacy. Keywords: Clostridium botulunium; Botulinum Neurotoxin Type F (BoNT/F) domains; cloning; recombinant protein expression; immunoreactivity DOI: http://dx.doi.org/10.3126/njb.v2i1.5634 Nepal Journal of Biotechnology Jan.2012, Vol.2(1): 1-1

Scenario-Driven Search for Pedestrians aimed at Triggering Non-Reversible Systems

Abstract-This paper presents the results of an innovative approach to pedestrian detection for automotive applications in which a non-reversible system is used; therefore the aim is to reach a very low false detection rate, ideally zero, by searching for pedestrians in specific areas only. The great advantages of such an approach are that pedestrian recognition is performed on limited image areas-therefore boosting its timing performance- and no assessment on the danger level is finally required before providing the result to either the driver or an on-board computer for automatic manoeuvres. This system has been extensively tested on two prototype vehicles equipped with one laserscanner, one camera, and brakeby-wire technology both in Italy and Korea; this paper describes the extensive tests and shows performance measurements. I

Redescription of Two Urostylid Ciliates (Ciliophora: Urostylida), Anteholosticha pulchra and Metaurostylopsis struederkypkeae from Korea

Two urostylid ciliates, Anteholosticha pulchra (Kahl, 1932) Berger, 2003 and Metaurostylopsis struederkypkeae Shao et al., 2008, new to Korea, were collected from the Yellow Sea and the East Sea, Korea, respectively. They were identified based on live observation and protargol impregnation. Taxonomical characters of A. pulchra are as follows: 190-300×30-55 μm size in vivo; contractile vacuole located on the left side of the posterior 1/4 of the cell; spherical-reddish granules at cirral bases and around dorsal bristles, somewhat sparsely distributed throughout the cell surface; four frontal and two frontoterminal cirri; four dorsal kineties; caudal cirri absent. Metaurostylopsis struederkypkeae is characterized as follows: 80-110×40-50 μm size in vivo; caudal cirri absent; two types of cortical granules: type 1, yellow-green arranged along the ventral cirral rows and dorsal kineties; type 2, small and reddish, with an irregular arrangement; four frontal, four to eight frontoterminal, and two to six transverse cirri; five to seven left and three to five right marginal rows. Sequences of small subunit ribosomal DNA were determined from both species, and pairwise distances with their relatives were analyzed

Morphology and Molecular Phylogeny of Pseudouroleptus jejuensis nov. spec., a New Soil Ciliate (Ciliophora, Spirotrichea) from South Korea

A new soil ciliate, Pseudouroleptus jejuensis, was discovered from Jeju Island, South Korea and described based on live observation, protargol impregnation, and SSU rRNA gene sequence analyses. Pseudouroleptus jejuensis differs from other congeneric species mainly by number of dorsal kineties (5 vs. 4). Based on our observation of late dividers, we confirm that the dorsal kinety anlage 3 forms 3 kineties (i.e., dorsal kineties 3–5), and the dorsal kinety anlagen 1–3 form 3–5/1–2/0 caudal cirri, respectively. Our gene trees support the assignment of this new species in Pseudouroleptus to full supporting values

Genome shotgun sequencing and development of microsatellite markers for gerbera (Gerbera hybrida H.) by 454 GS-FLX

The objective of this research was to develop and characterize microsatellite markers for gerbera. We used shotgun sequencing with Roche 454 GS-FLX Titanium technology to identify microsatellite loci in gerbera genomic DNA (Gerbera hybrida). The total length of non-redundant sequences obtained was 22,527,019 bp, which consisted of 3,085 contigs and 28,249 singletons. We assembled 61,958 reads into 3,085 contigs, of which 114 (3.70%) contained microsatellite repeats. The average G+C content was 39.3%. Functional annotation to known sequences yielded 14.7% unigenes in the ‘Raon’ cultivar. Analysis of the gerbera genome DNA (‘Raon’) general library showed that sequences of (AT), (AG), (AAG) and (AAT) repeats appeared most often, whereas (AC), (AAC) and (ACC) were the least frequent. Primer pairs were designed for 80 loci. Only eight primer pairs produced reproducible polymorphic bands in the 28 gerbera accessions analyzed. A total of 30 alleles were identified from the eight polymorphic SSR loci, with two to eight alleles per locus (average level of 3.75). These markers will be useful for investigating genetic diversity and differentiation in gerbera. Keywords: Genetic diversity, genomics, microsatellite isolation, pyrosequencing, SSRs. African Journal of Biotechnology Vol. 11(29), pp. 7388-7396, 10 April, 201