Molecular subtyping methods have previously shown that there is a nonrandom distribution of Escherichia coli O157:H7 strains among clinical and nonclinical isolates. Two examples include the lineage-specific polymorphism assay (LSPA) and clade typing assay. The clade typing method was previously used to identify a phylogenetic group of E. coli O157:H7 designated clade 8, which is believed to be more virulent than non-clade 8 isolates, and the LSPA previously indicated that clade 8 isolates are LSPA genotype 211111. Published screens have suggested that LSPA 211111 comprise anywhere from 3.9% to greater than 46% of clinical isolates. To determine the prevalence of such isolates within Pennsylvania, we applied LSPA and screened 52 clinical isolates. We found that 31% of isolates were LSPA 211111 and that 13/16 of these could be classified as clade 8. A rapid polymerase chain reaction screen for clade 8 isolates was developed and shown to have a specificity and sensitivity of 0.92 and 1.0, respectively. Polymerase chain reaction screens indicated that all isolates carried hlyA and eaeA and that all but one of the isolates carried katP. The most common LSPA genotype seen within our collection was 111111, and 29 of 30 of these carried both stx1 and stx2. Clade 8 isolates were more diverse, with four different Shiga toxin profiles observed. We conclude that E. coli O157:H7 of LSPA 211111 and clade 8 are common to clinical isolates in Pennsylvania and suggest that further studies are needed to determine whether their prevalence is increasing as observed elsewhere.

A total number of 1105 cases with viral conjunctivitis, mainly epidemic kerato-conjunctivitis (EKC) consisting of 354 cases from Sapporo, Japan, of 628 from Kaohsiung, Taiwan and of 123 from Busan, Korea, encountered during the periods, 1980-81 and/or 1983, were studied aetiologically and epidemiologically. Patients were aged from 27 days to 88 years with a peak in the 20-29 year age group. Aetiological diagnoses were established in 610 cases (55%), consisting of 536 cases (49%) caused by adenoviruses and of 74 (7%) by EV 70. The most frequently detected agent was Ad 8 (57%), followed by EV 70 (12%), Ad 3 (9%) and Ad 19 (7%). The aetiological profiles of viral conjunctivitis were generally similar in three cities of East Asia. EKC (70%) was mainly caused by Ad 8, Ad 19 and Ad 37, but AHC (13%) by EV 70 and PCF (5%) by Ad 3 and Ad 11. Ad 19 and Ad 37 isolates in three cities were compared with the cleavage patterns with restriction endonucleases, BamHl, Smal, Sall and Hindlll. All of the Ad 19 isolates tested were identical to Ad 19a reported by Wadell et al, and all but three isolates of Ad 37 were identical to the Ad 37 prototype prevalent in Europe and the US. The three isolates of Ad 37, different from the prototype in cleavage pattern with Hindlll, designated as Ad 37A, were detected in Sapporo and Kaohsiung in 1980. From the cleavage patterns with four restriction endonucleases, Hindlll, BamHl, Sa1l and Sstl, the Ad 8 isolates in 1983 were divided into three subtypes, which were associated with the cities isolated.(ABSTRACT TRUNCATED AT 250 WORDS)

The Society for Translational Medicine and The Chinese Society for Thoracic and Cardiovascular Surgery conducted a systematic review of the literature in an attempt to improve our understanding in the postoperative management of chest tubes of patients undergoing pulmonary lobectomy. Recommendations were produced and classified based on an internationally accepted GRADE system. The following recommendations were extracted in the present review: (I) chest tubes can be removed safely with daily pleural fluid of up to 450 mL (non-chylous and non-sanguinous), which may reduce chest tube duration and hospital length of stay (2B); (II) in rare instances, e.g., persistent abundant fluid production, the use of PrR <0.5 when evaluating fluid output to determine chest tube removal might be beneficial (2B); (III) it is recommended that one chest tube is adequate following pulmonary lobectomy, except for hemorrhage and space problems (2A); (IV) chest tube clearance by milking and stripping is not recommended after lung resection (2B); (V) chest tube suction is not necessary for patients undergoing lobectomy after first postoperative day (2A); (VI) regulated chest tube suction [-11 (-1.08 kPa) to -20 (1.96 kPa) cmHO depending upon the type of lobectomy] is not superior to regulated seal [-2 (0.196 kPa) cmHO] when electronic drainage systems are used after lobectomy by thoracotomy (2B); (VII) chest tube removal recommended at the end of expiration and may be slightly superior to removal at the end of inspiration (2A); (VIII) electronic drainage systems are recommended in the management of chest tube in patients undergoing lobectomy (2B).

The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.

As one of the branched-chain amino acids, L-valine is an essential nutrient for most mammalian species. In this study, the L-valine producer Corynebacterium glutamicum ΔppcΔaceEΔalatΔpqo was first constructed. Additionally, an improved biosensor based on the Lrp-type transcriptional regulator and temperature-sensitive replication was built. Then, the C. glutamicum strain was mutagenized by atmospheric and room temperature plasma. A sequential three-step procedure was carried out to screen L-valineproducing strains, including the fluorescence-activated cell sorting (FACS), 96-well plate screening, and flask fermentation. The final mutant HL2-7 obtained by screening produced 3.20 g/L of L-valine, which was 21.47% higher than the titer produced by the starting strain. This study demonstrates that the L-valine-producing mutants can be successfully isolated based on the Lrp sensor system in combination with FACS screening after random mutagenesis.