Surveillance of tuberculosis treatment outcomes of Singapore citizens and permanent residents, 2002-2011.

SETTING: Singapore, which had a tuberculosis (TB) incidence rate of 41 per 100,000 resident population in 2011. OBJECTIVE: To report the outcomes of Singapore citizens and permanent residents treated for TB from 2002 to 2011. METHODS: A computerised treatment surveillance module (TSM) was launched in 2001 to track the progress and outcome of TB patients nationally. Physicians were required to submit an electronic or paper return for every patient at each clinic visit. Treatment adherence, drugs prescribed, treatment delivery mode and final outcome, specified as 'completed treatment', 'lost to follow-up', 'death', 'transferred out', 'permanent cessation of treatment' and 'still on treatment/no final outcome', were captured. Quarterly cohort outcomes at 12-15 months after starting treatment were combined to generate annual treatment outcomes. RESULTS: Treatment completion rates increased from 73.4% to 82.8%. The proportion of patients lost to follow-up decreased from 3.4% to 1.7%, while that of patients still on treatment or with no final outcome decreased from 10.5% to 4.4%. The death rate ranged between 10.2% and 11.7%; the majority were not attributed to TB. CONCLUSION: TB treatment completion among Singapore citizens and permanent residents has improved since 2002 as the likely result of the TSM and other initiatives introduced over the past decade.

Viral agents associated with acute gastroenteritis in Seoul, Korea.

BACKGROUND: The present study was carried out to describe the epidemiologic characteristics of viral gastroenteritis and determine the phylogenetic composition of norovirus strains detected in hospitalized children with acute gastroenteritis in Seoul, Korea. METHODS AND RESULTS: In total, 10,603 stool samples were collected from 2004 to 2008 and tested by RT-PCR or ELISA. In 4,170 (39.3%) samples at least one viral pathogen was present. Rotavirus (RoV) (1,864, 17.5%) was found to be the causative agent followed by norovirus (NoV) (1,845, 17.4%), human adenovirus (HAdV) (266, 2.5%), human astrovirus (HAstV) (194, 1.8%), and sapovirus (SV) (1, 0.009%). Five GI genotypes (GI-1, GI-3, GI-4, GI-8, and GI-9) and eight GII genotypes (GII-2, GII-3, GII-4, GII-6, GII-7, GII-12, GII-16, and GII-17) of NoV were identified in acute gastroenteritis patients in 2008. CONCLUSIONS: The genetic characteristics of norovirus and the epidemiologic patterns of a viral pathogen from acute gastroenteritis patients may give potentially effective data for epidemiological studies in Seoul, Korea.

Enhanced degradation and toxicity reduction of dihexyl phthalate by Fusarium oxysporum f. sp. pisi cutinase.

AIMS: This research aims to investigate the efficiency of two lipolytic enzymes--fungal cutinase and yeast esterase--upon the biodegradation of dihexyl phthalate (DHP). METHOD AND RESULTS: During the enzymatic degradation of DHP dissolved in methanol, several degradation products were detected and their time-course changes were monitored using GC/MS. The DHP-degradation rate of cutinase was surprisingly high; i.e. almost 70% of the initial DHP (500 mg l(-1)) was decomposed within 4.5 h. Although the same amount of esterase was employed, more than 85% of the DHP remained after 3 days. Almost all the DHP was converted by cutinase into 1,3-isobenzofurandione (IBF), whereas hexyl methyl phthalate and IBF were abundantly produced by esterase. In addition, the toxicities of the DHP-degraded products by esterase were evaluated using various recombinant bioluminescent bacteria, which caused oxidative and protein damage, whereas the hydrolysis products from cutinase never caused any cellular damage in the methanol-containing reaction system. CONCLUSIONS: Cutinase starts to act as a DHP-degrader much earlier and faster than esterase, with high stability in ester-hydrolytic activity, therefore a plausible approach to the practical application of cutinase for DHP degradation in the DHP-contaminated environments may be possible. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes the enhanced degradation and detoxification of DHP using Fusarium oxysporum f. sp. pisi cutinase.

Molecular cloning of kpcA gene encoding a Kex2p-like endoprotease from Aspergillus nidulans.

We cloned and sequenced a gene, kpcA (Kex2p-like proprotein convertase A), from a genomic library of Aspergillus nidulans. The kpcA gene encodes an 820-residue protein, named KpcA, which contains a putative subtilisin-like catalytic domain (residues 136-466) homologous to that of the subtilisin serine protease family. KpcA shows 56, 73, and 47% amino acid identities with Saccharomyces cerevisiae Kex2p, Aspergillus niger KexB, and mouse furin within the subtilisin-like catalytic domain, respectively. The sequences around the proposed active site Asp, His, and Ser residues of KpcA are similar to those of other Kex2p family members. The KpcA mRNA transcript with an expected size of approximately 2.8 kb was detected in A. nidulans. The substrate specificity of KpcA, expressed in CHO cells, is similar to that of A. niger KexB and yeast Kex2p. We conclude that KpcA is a resident Kex2p-like proprotein that processes endoprotease in A. nidulans.

The nsdD gene encodes a putative GATA-type transcription factor necessary for sexual development of Aspergillus nidulans.

The ability to reproduce both sexually and asexually is one of the characteristics of the homothalic ascomycete Aspergillus nidulans. Unlike the other Aspergillus species, A. nidulans undergoes sexual development that seems to be regulated by internal and external stimuli. To begin to understand the sexual reproduction of A. nidulans we previously isolated and characterized several NSD (never in sexual development) mutants that failed to produce any sexual reproductive organs, and identified four complementation groups, nsdA, nsdB, nsdC, and nsdD. The nsdD gene has been isolated, and it is predicted to encode a GATA-type transcription factor with the type IVb zinc finger DNA-binding domain. The mRNA of the nsdD gene started to accumulate in the early phase of vegetative growth, and the level increased as sexual development proceeded. However, it decreased during asexual sporulation and no nsdD mRNA was detected in conidia. Deletion of nsdD resulted in no cleistothecia (fruiting bodies) formation, even under the conditions that preferentially promoted sexual development, indicating that nsdD is necessary for sexual development. In contrast, when the nsdD gene was over-expressed, sexual-specific organ (Hülle cell) was formed even in submerged culture, which normally completely blocked sexual development, and the number of cleistothecia was also dramatically increased on solid medium. These results lead us to propose that the nsdD gene functions in activating sexual development of A. nidulans. Multiple copies of the nsdD gene could suppress nsdB5 and veA1, indicating that either nsdD acts downstream of these genes or possibly functions in overlapping pathway(s).

Isolation and characterization of cultured human periodental ligament fibroblast-specific cDNAs.

The molecular mechanisms that control the function of periodontal ligament (PDL) fibroblasts remain unclear. We speculated that the character of differentiating PDL fibroblasts is defined by the altered expansion of specific genes not found in neighboring gingival fibroblasts in the periodontium. To expand this set, subtractive hybridization was applied between cultured human PDL and gingival fibroblasts to identify genes differentially expressed in PDL. Consequently five candidate clones, PDLs (periodontal ligament specific) 5, -17, -22, -25, and -31 were identified and characterized by homology search, Northern analysis, and in situ hybridization. Although the mRNAs of these clones were expressed by bone marrow cells and rarely by gingival fibroblasts, the highest expression was detected in the PDL cells, which were uniformly distributed throughout the whole PDL. Amongst the five candidate clones, we focused on PDLs17, because it is a hypothetical protein whose biological function has not been reported yet in the database. Polyclonal antiserum raised against PDLs17 peptide was made, and stained the PDL fibroblasts, osteoblast-like cells and stromal cells in the bone marrow, but not gingival fibroblasts. The results suggest that clones, PDLs5, -17, -22, -25, and -31 may be used as PDL fibroblast-specific markers, and that PDLs17 could act as an important factor in the differentiation process of PDL fibroblasts.

Differential expression of house-keeping genes of Aspergillus nidulans during sexual development.

The rpl3 gene and the rpl37 gene for Aspergillus nidulans ribosomal protein L3 (RPL3) and RPL37, which were identified as located on chromosome I and chromosome III, respectively, were isolated from chromosome-specific cosmid libraries. The nucleotide sequences of both of the rpl3 gene and the rpl37 gene identified the ORFs of 392 amino acids and 92 amino acids, respectively. Both of the two genes were present in a single copy. The expression of both genes together with two other house-keeping genes, the rps16 gene for RPS16 and the gene for gamma-actin, was analyzed during sexual development. All four genes showed nearly identical expression patterns in that each gene expression reached its maximum after 2 h, decreased thereafter, and increased again after 30-40 h of induction of sexual development.

Biceps load test: a clinical test for superior labrum anterior and posterior lesions in shoulders with recurrent anterior dislocations.

We describe the biceps load test for evaluating the integrity of the superior glenoid labrum in shoulders with recurrent anterior dislocations. With the shoulder in an abducted, externally rotated position and the forearm supinated, active flexion of the elbow against resistance relieves the discomfort of a standard apprehension test for anterior shoulder instability. A group of 75 patients with proven unilateral anterior shoulder dislocations were prospectively examined in a double-blind fashion with arthroscopic examination and the biceps load test. Sixty-three patients had a negative test and 62 of these had an intact biceps tendon-superior labrum complex; the remaining patient had a type II superior labral anterior and posterior lesion. Twelve patients had positive tests, and 10 had superior labral lesions; the other 2 patients had intact superior labra. Therefore, the biceps load test revealed a sensitivity of 90.9%, a specificity of 96.9%, a positive predictive value of 83%, a negative predictive value of 98%, and a kappa coefficient of 0.846.

The uvsI gene of Aspergillus nidulans required for UV-mutagenesis encodes a homolog to REV3, a subunit of the DNA polymerase zeta of yeast involved in translesion DNA synthesis.

Defects in the uvsI gene of Aspergillus nidulans resulted in high UV sensitivity and reductions of spontaneous and UV-induced reversion of certain alleles, uvsl;uvsA double mutants exhibited high methyl methane sulfonate (MMS)-sensitivity in contrast to the slight sensitivity of the component single mutants. Using such a double mutant as recipient, a clone complementing uvsI501 has been isolated from a chromosome III specific library. The deduced amino acid sequence from the 1.1-kb sequenced region, a part of the 5.2-kb DNA fragment showing uvsI-complementing activity, had a 62% identity with REV3 of yeast. Disruptants of the cloned gene demonstrated the same level of sensitivity to UV light as uvsI and failed to complement uvsI501 in heterozygous diploids.

Immunohistochemical expression of extracellular matrix components of normal and healing periodontal tissues in the beagle dog.

Periodontal regeneration requires formation of periodontal tissues lost due to periodontal disease. To better understand the formation of new periodontal tissues during periodontal repair and regeneration, immunohistochemical expression of extracellular matrix components of normal as well as healing periodontal tissues was evaluated and compared using the avidin-biotin complex immunohistochemical technique. For this purpose, horizontal furcation defects were created around mandibular P2 and P4 of 6 dogs after extraction of P1 and P3. The root surfaces were conditioned with citric acid and expanded polytetrafluoroethylene (ePTFE) membranes were placed and retained 0.5 mm above the cemento-enamel junction. The mucoperiosteal flaps were sutured in a coronal position. Two animals were sacrificed at 2, 4, and 8 weeks, and mesio-distal tissue slices containing normal or healing periodontal tissues were demineralized, dehydrated, and embedded in paraffin. Immunohistochemical localization of type I collagen (CI), fibronectin (FN), secreted protein, acidic and rich in cysteine (SPARC), vitronectin (VN), and bone sialoprotein (BSP) was performed on 6 microns thick sections. Morphological results demonstrated that at 2 weeks after defect creation, lesions were filled primarily with granulation tissue which was gradually replaced by newly-formed fibrous connective tissue, periodontal ligament (PDL), cementum, and bone between 4 and 8 weeks. The results of immunohistochemical study revealed that at 2 weeks the granulation tissue, especially in the intercellular spaces of inflammatory cells, was intensively stained for FN and VN. At 4 and 8 weeks, staining for CI, FN, and VN was found in fibrous connective tissue, the newly-formed PDL, cementum, and osteoid. Further the attachment zone of the PDL collagen fibers to cementum showed intense staining for FN. Immunostaining for SPARC was positive in the new PDL, cementum, and bone, while staining for BSP was restricted to the new cementum and bone. Interestingly, the PDL, especially in areas adjacent to active bone formation, demonstrated intense staining for BSP. However, fibrous connective tissue and PDL proper were unstained for BSP. These results indicate that FN and VN are involved in the early stages of periodontal repair, and periodontal regeneration is achieved through formation of periodontal tissues that are composed of different matrix components specific to different types of periodontal tissues.

Periodontal regeneration in class III furcation defects of beagle dogs using guided tissue regenerative therapy with platelet-derived growth factor.

We developed an effective regenerative therapy, referred to as platelet-derived growth factor-BB (PDGF-BB)-modulated guided tissue regenerative (GTR) therapy (P-GTR), capable of achieving periodontal regeneration of horizontal (Class III) furcation defects in the beagle dog. To determine its efficacy, repair and regeneration of horizontal furcation defects by P-GTR therapy and GTR therapy were compared. Chronically inflamed horizontal furcation defects were created around the second (P2) and fourth mandibular premolars (P4). After demineralization of the root surfaces with citric acid, the surfaces of left P2 and P4 were treated with PDGF-BB (P-GTR therapy) and those of contralateral teeth were treated with vehicle only (GTR therapy). Periodontal membranes were placed and retained 0.5 mm above the cemento-enamel junction for both groups. The mucoperiosteal flap was sutured in a coronal position and plaque control was achieved by daily irrigation with 2% chlorhexidine gluconate. At 5, 8, and 11 weeks, two animals each were sacrificed by perfusion with 2.5% glutaraldehyde through the carotid arteries, and the lesions were sliced mesio-distally, demineralized, dehydrated, and embedded. Periodontal healing and regeneration after GTR and P-GTR therapy were compared by histomorphometric as well as morphological analysis. Morphometric analysis for each time period was performed on the pooled samples of P2 and P4. Five weeks after both therapies, the lesions were filled primarily by tissue-free area, epithelium, inflamed tissue, and a small amount of newly formed fibrous connective tissue. At 8 and 11 weeks after P-GTR therapy, there was a statistically greater amount of bone and periodontal ligament formed in the lesions. The newly formed bone filled 80% of the lesion at 8 weeks and 87% at 11 weeks with P-GTR therapy, compared to 14% of the lesion at 8 weeks and 60% at 11 weeks with GTR therapy. Also, with P-GTR therapy there was less epithelium and tissue-free area, less inflamed tissue, and less connective tissue. Morphological analysis indicated that the defects around P2 revealed faster periodontal repair and regeneration than those around P4. While the lesions around P2 were effectively regenerated by 11 weeks even after GTR therapy, those around P4 failed to regenerate. On the other hand, P-GTR therapy further promoted periodontal repair and regeneration so that at 8 weeks the lesions around P2 and P4 demonstrated complete and nearly complete regeneration, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Efficacy of polyadenylic.polyuridylic acid in the treatment of chronic active hepatitis B.

In order to assess whether polyadenylic.polyuridylic acid [poly(A).poly(U)] can be used as a new therapeutic agent for the treatment of chronic hepatitis B, 19 patients with histologically proven chronic active hepatitis B were injected intravenously with 100-150 mg of poly(A).poly(U) weekly for six weeks. Changes in alanine aminotransferase (ALT) levels, 2',5'-oligoadenylate synthetase (2'.5'-AS) activities and HBV markers were sequentially checked during and after treatments. Serum ALT levels were decreased gradually and 2'.5'-AS activities were significantly increased after initiation of poly(A).poly(U) injections. At the end of this trial (24th week) we have observed the normalizations of elevated ALT levels in 14 (73.7%), negative conversion of HBeAg in 11 (57.9%) and loss of HBV-DNA in 12 out of 19 patients (63.1%). Complete responses which had both normalization of ALT levels and negative conversion of HBeAg were noted in 11 patients (57.9%) and partial responses showing either normalization of ALT levels or negative conversion of HBeAg alone were in four out of 19 patients (21.1%). No notable adverse effects were observed during the treatments and follow-up period. It can be concluded that poly(A).poly(U) seems to be effective in the treatment of chronic active hepatitis B and has an advantage of being free of significant side effects.