A tuberculosis school outbreak in China, 2018: reaching an often overlooked adolescent population.

Adolescents have been largely neglected from tuberculosis control efforts. In low- to medium burden settings much of the tuberculosis burden in this age group occurs from school outbreaks. We report on a large tuberculosis outbreak in adolescents from a boarding high school in Jiangsu Province, China. From March to June 2018, a tuberculosis outbreak occurred in a boarding high school. We conducted an outbreak investigation involving clinical diagnostic tests and molecular analysis to determine the outbreak origin. Cases were detected through symptom screening, tuberculin skin testing (TST), chest radiography, sputum smear, solid sputum culture and GeneXpert MTB/RIF. Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) genotyping and spoligotyping methods were performed on Mycobacterium tuberculosis (M. tuberculosis) isolates to identify the outbreak origin. A total of 845 students and 131 teachers/staff attended a TST screening for tuberculosis infection. The prevalence of elevated tuberculin reactions at ≥5, ≥10 and ≥15 mm was 12.19% (119/976), 6.35% (62/976) and 3.28% (32/976), respectively. Radiographic abnormalities were present in 5.73% (56 of 976) individuals, 40 students and 16 teachers/staff. Of these, 12 students were diagnosed with confirmed tuberculosis. In total, 14 students (two index cases and 12 confirmed cases) were diagnosed and reported in the tuberculosis outbreak, an attack rate of 1.7% (14/847) among students (two index cases and 845 screened students). Results from MIRU-VNTR typing and spoligotyping analyses demonstrated that three M. tuberculosis strains belong to the Beijing family with corresponding MIRU-VNTR alleles. This school-based tuberculosis outbreak among adolescents demonstrates that transmission among individuals in this age group is common and must be prioritised. It suggests that identifying and timely diagnosis of smear-positive cases, especially in the early phase of outbreaks, is the key to preventing further spread among close contacts.

[Progress in research of bacillus calmette-guerin vaccination and repeated vaccination].

Tuberculosis (TB) is a serious infectious diseases threating human health, bacillus balmette-guerin vaccine (BCG) is the only available TB vaccine now, neonatal vaccination can significantly reduce the incidence and death of tuberculosis. However, due to its limited protection period, one dose vaccination after birth does not have a protective effect for adolescents and adults. Therefore, how to reduce the prevalence of TB in adolescents and adults effectively is essential for TB prevention and control. In this paper, we reviewed the literature from PubMed, CNKI, and Wanfang database to analyze and summarize the characteristics of BCG vaccine, immune effects and immunity endurance, the effects of BCG vaccination and repeated BCG vaccination in adolescents adults and discuse the change of attitude and trends of BCG use in the three documents issued by the World Health Organization on position of BCG.

Resveratrol transcriptionally regulates miRNA-18a-5p expression ameliorating diabetic nephropathy via increasing autophagy.

OBJECTIVE: To investigate the effects of resveratrol on autophagy in the chronically diabetic nephropathy and to study the effects of the different expression of microRNAs after resveratrol (RSV) treated in db/db mice (diabetic mice). MATERIALS AND METHODS: Db/m (non- diabetic) and db/db mice were randomly divided into intra gastric RSV treatment group or control group. Renal tissues were prepared for HE/PAS staining. In vitro, mouse podocytes cell lines were grown in different mediums with different dose of resveratrol treatment. microRNA (miRNA) gene chips assay was performed for differentially expressed miRNAs screening. Western blot was used to detect protein levels. RESULTS: In vivo, RSV significantly decreased urinary albumin, serum creatinine, mesangial area and glomerular size in db/db mice. After RSV treatment, LC3-II/LC3-I and synaptopodin were increased while cleaved-caspase 3 was decreased in kidney tissues. In vitro, podocytes treated with RSV exhibited significantly increased LC3-II/LC3-I and decreased cleaved caspase 3. Moreover, this effect of RSV can be enhanced by rapamycin (RAPA, an activator of autophagy) but partially reversed by 3-MA (an autophagy inhibitor). Further, we found that miR-18a-5p was significantly upregulated after RSV treatment in db/db mice. Overexpression of miR-18a-5p in podocytes resulted in significant inhibition of cleaved-caspase 3 protein, and increased the ratio of LC3-II/LC3-I. Dual luciferase report assay validated that Atactic telangiectasis mutation (ATM) was a target of miR-18a-5p. In podocytes, downregulation of cleaved caspase 3 and the enhanced ratio of protein LC3-II/LC3-I were detected in cells transfected with ATM siRNA. CONCLUSIONS: Role of miRNA-18a-5p in the regulation of autophagy via targeting ATM may represent a promising therapeutic target for preventing and attenuating diabetic nephropathy.

The impact of acute bouts of exercise on circulating cardiac natriuretic peptide and left ventricular function in untrained healthy young subjects.

BACKGROUND: To examine the impact of a single bout of exercise on cardiac function and the blood level of brain natriuretic peptide (BNP) in healthy subjects. METHODS: Thirty untrained, healthy young (aged between 20 and 29 years) volunteers (15 men and 15 women) were prospectively recruited. The plasma N-terminal pro-B-type BNP (NT-proBNP) level was checked, cardiac function was assessed using echocardiography in baseline conditions, and the values were compared with those obtained after treadmill exercise according to the Bruce protocol. RESULTS: Left ventricular (LV) ejection fraction decreased after exercise in men, but not in women. Parameters of LV diastolic function were likely to change unfavorably in both sexes after exercise. NT-proBNP levels were significantly elevated by exercise in both sexes (from 22.1±18.3 to 24.7±20.2 pg/mL, P=0.006 in men; from 41.1±21.5 to 46.5±24.9 pg/mL, P<0.001 in women). Changes in NT-proBNP levels were significantly associated with those in E wave velocity, E wave deceleration time, E/e' and E/propagation velocity (Vp) in men (P<0.05 for each) even after controlling for the effects of blood pressure and heart rate. In women, there were no significant associations between changes in parameters of LV diastolic function and those in NT-proBNP levels. CONCLUSIONS: LV systolic and diastolic functions may deteriorate, and proBNP levels could be elevated even by a single bout of treadmill exercise in untrained young subjects. Dynamic change in LV diastolic function is a major stimulus to NT-proBNP release following exercise in men but not in women.

Comparison of four facial muscles, orbicularis oculi, corrugator supercilii, masseter or mylohyoid, as best predictor of good conditions for intubation: a randomised blinded trial.

CONTEXT: Monitoring of facial muscles after neuromuscular blockade can give an early indication of respiratory muscle readiness for tracheal intubation. OBJECTIVE: To assess which facial muscle, the orbicularis oculi, corrugator supercilii, masseter or the mylohyoid, is the best predictor of readiness for intubation after rocuronium. DESIGN: Prospective, randomised, blinded trial. SETTING: Single centre: Seoul, Korea, from August 2012 to November 2012. PATIENTS: Two hundred and eighty-eight patients aged 22 to 64 years were randomised to one of eight study groups: orbicularis oculi, corrugator supercilii, masseter and mylohyoid for rocuronium 0.6 or 1.2 mg kg. INTERVENTION: The maximum twitch depression at the eyelid (orbicularis oculi), the superciliary arch (corrugator supercilii), the cheek (masseter) and the submental triangle (mylohyoid) was assessed after rocuronium 0.6 and 1.2 mg kg. Endotracheal intubation was performed after maximal neuromuscular blockade, and intubating conditions were appraised. MAIN OUTCOME MEASURES: The onset time of rocuronium and the quality of the intubation conditions were assessed. RESULTS: The onset times in the orbicularis oculi, corrugator supercilii and masseter were significantly faster than that in the mylohyoid (P < 0.001). 'Clinically acceptable' intubation conditions were significantly enhanced in the mylohyoid (94%) compared with those in the orbicularis oculi (80%) and masseter (78%) after rocuronium 0.6 mg kg (P < 0.05), and no difference with corrugator supercilii (92%). Despite differences in onset time of orbicularis oculi and masseter compared to mylohyoid (P < 0.05), intubating conditions were similar among the four muscles after rocuronium 1.2 mg kg. CONCLUSION: Following rocuronium 0.6 mg kg at similar depths of anaesthesia, the monitoring of the corrugator supercilii provided the best balance of a shorter onset time while maintaining 'clinically acceptable' intubation conditions. TRIAL REGISTRATION: IRB File No.: HYUH 2012-07-009.

Insertion sequence- and tandem repeat-based genotyping techniques for Xanthomonas citri pv. mangiferaeindicae.

Molecular fingerprinting techniques that have the potential to identify or subtype bacteria at the strain level are needed for improving diagnosis and understanding of the epidemiology of pathogens such as Xanthomonas citri pv. mangiferaeindicae, which causes mango bacterial canker disease. We developed a ligation-mediated polymerase chain reaction targeting the IS1595 insertion sequence as a means to differentiate pv. mangiferaeindicae from the closely related pv. anacardii (responsible for cashew bacterial spot), which has the potential to infect mango but not to cause significant disease. This technique produced weakly polymorphic fingerprints composed of ≈70 amplified fragments per strain for a worldwide collection of X. citri pv. mangiferaeindicae but produced no or very weak amplification for pv. anacardii strains. Together, 12 tandem repeat markers were able to subtype X. citri pv. mangiferaeindicae at the strain level, distinguishing 231 haplotypes from a worldwide collection of 299 strains. Multilocus variable number of tandem repeats analysis (MLVA), IS1595-ligation-mediated polymerase chain reaction, and amplified fragment length polymorphism showed differences in discriminatory power and were congruent in describing the diversity of this strain collection, suggesting low levels of recombination. The potential of the MLVA scheme for molecular epidemiology studies of X. citri pv. mangiferaeindicae is discussed.

Diphenhydramine modulates cytokines and induces apoptosis in experimental acute pancreatitis.

In this study, we investigated the therapeutic potential of diphenhydramine (DPH), a H(1) receptor antagonist, on taurocholate-induced acute pancreatitis and the underlying mechanisms involved. Rats were randomly divided into sham-operated, model, DPH-treated, octreotide-treated and the DPH plus octreotide combination therapy groups (n = 30 per group). Animals were sacrificed 3, 6 and 24 h after modeling and drug administration (n = 10 per time point) and sera, pancreas and lungs were harvested for further studies. DPH and octreotide monotherapy relieved histopathological injuries in multiple organs when compared to the model group. Combination therapy (DPH + octreotide) demonstrated better therapeutic potential than monotherapy. Data indicated that combination therapy had a better ability to reduce average mortality rates in rats, decrease the number of inflammatory cells, attenuate necrosis, upregulate the levels of amylase, TNF-alpha and IL-8 and downregulate the levels of IL-10 in the serum. Moreover, enhanced expression of Bax in the pancreas and lung were recorded suggesting a pro-apoptotic mechanism involved in the therapeutic potential of DPH. Our study demonstrated the therapeutic potential of DPH in acute pancreatitis and suggested a novel strategy for clinical management of this disease.

Polyphasic characterization of xanthomonads pathogenic to members of the Anacardiaceae and their relatedness to species of Xanthomonas.

We have used amplified fragment length polymorphism (AFLP), multilocus sequence analysis (MLSA) and DNA-DNA hybridization for genotypic classification of Xanthomonas pathovars associated with the plant family Anacardiaceae. AFLP and MLSA results showed congruent phylogenetic relationships of the pathovar mangiferaeindicae (responsible for mango bacterial canker) with strains of Xanthomonas axonopodis subgroup 9.5. This subgroup includes X. axonopodis pv. citri (synonym Xanthomonas citri). Similarly, the pathovar anacardii, which causes cashew bacterial spot in Brazil, was included in X. axonopodis subgroup 9.6 (synonym Xanthomonas fuscans). Based on the thermal stability of DNA reassociation, consistent with the AFLP and MLSA data, the two pathovars share a level of similarity consistent with their being members of the same species. The recent proposal to elevate X. axonopodis pv. citri to species level as X. citri is supported by our data. Therefore, the causal agents of mango bacterial canker and cashew bacterial spot should be classified as pathovars of X. citri, namely X. citri pv. mangiferaeindicae (pathotype strain CFBP 1716) and X. citri pv. anacardii (pathotype strain CFBP 2913), respectively. Xanthomonas fuscans should be considered to be a later heterotypic synonym of Xanthomonas citri.

First Report of Xanthomonas cucurbitae Causing Bacterial Leaf Spot of Watermelon in the Seychelles.

Bacterial leaf spot of cucurbits caused by Xanthomonas cucurbitae (4) can be a harmful disease of several cucurbit species in tropical environments, mainly within the Cucumis, Cucurbita, and Citrullus genera. The bacterium induces angular, water-soaked leaf spots, which sometimes become necrotic and have a chlorotic halo. Scab-like lesions on fruit can also be observed (2). Water-soaked, angular leaf lesions were collected from approximately 15 watermelon plants (Citrullus lanatus) in a production field located in Mahé, Seychelles in 2003. Yellow-pigmented Xanthomonas-like bacterial colonies were isolated on KC semiselective medium (yeast extract 7 g, peptone 7 g, glucose 7 g, agar 18 g, distilled water 1,000 ml, propiconazole 20 µg ml-1, cephalexin 40 mg liter-1, and kasugamycin 20 mg liter-1) from all isolation attempts (3). Amplified fragment length polymorphism (AFLP) analysis was performed on four watermelon strains together with reference strains of Xanthomonas cucurbitae (LMG 690 [type strain] and LMG 8663) and the type strain of all other valid Xanthomonas species using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (1). The four strains from watermelon showed identical fingerprints and were most closely related to X. cucurbitae. One strain from diseased watermelon (JZ88-1) was further analyzed by MultiLocus Sequence Analysis (MLSA) using portions of three housekeeping genes (atpD, dnaK, and gyrB) as described previously (1). This strain displayed a very high relatedness (99.8 and 98.9% with strain LMG 690 and LMG 8663, respectively) to the two reference strains of X. cucurbitae. AFLP and MLSA were useful for identifying strains at the species level that were consistent with previous results (1). Bottle-gourd (Lagenaria siceraria), pumpkin (Cucurbita maxima), squash cv. Aurore (Cucurbita pepo), cucumber cv. L-04 (Cucumis sativus), cantaloupe melon cv. Cezanne (Cucumis melo), and watermelon cv. Fou-nan (C. lanatus) leaves were infiltrated (10 inoculation sites per leaf and three replicates) with bacterial suspensions (JZ88-1, LMG 690 and LMG 8663) containing approximately 1 × 105 CFU ml-1 (approximately 1 × 102 CFU per inoculation site). Typical water-soaked lesions that developed into necrotic spots were observed 6 to 8 days after inoculation for all inoculated strains on all inoculated plant species. One month after inoculation, Xanthomonas was recovered from lesions and population sizes determined on KC semiselective medium (3) ranging from 1 × 106 to 9 × 106 CFU per lesion were typical of a compatible interaction. Bacterial leaf spot has appeared sporadically in Mahé, Seychelles since 2003, most often with limited incidence. However, growers need to be aware of the potential negative effect of this disease in tropical environments. References: (1) N. Ah-You et al. Int. J. Syst. Evol. Microbiol. 59:306, 2009. (2) J. F. Bradbury. Page 309 in: Guide to Plant Pathogenic Bacteria. CAB International, Slough, UK, 1986. (3) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (4) L. Vauterin et al. Int. J. Syst. Bacteriol. 45:472, 1995.

First Report in the Seychelles of Xanthomonas axonopodis Genetic Cluster 9.2 Causing Bacterial Leaf Spot of Avocado.

Small, black, angular leaf lesions, which sometimes coalesced, were collected from avocado (Persea americana Miller) leaves in a government nursery located at Grand Anse, Mahé, Seychelles archipelago in 2003. Patterns of diseased plants were highly clustered, suggesting local dispersal in the nursery. Yellow-pigmented Xanthomonas-like bacterial colonies were isolated on KC semiselective medium (3). Amplified fragment length polymorphism (AFLP) analysis was performed on two avocado strains together with reference strains of the genetic clusters of Xanthomonas axonopodis (4) and the type strain of all other valid Xanthomonas species using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (1). The two avocado strains showed identical fingerprints and were closely related to X. axonopodis genetic cluster 9.2 (4). One strain, JZ103-1, was further analyzed by MultiLocus Sequence Analysis (MLSA) using portions of three housekeeping genes (atpD, dnaK, and gyrB) as described previously (1). MLSA data confirmed that the xanthomonad associated with avocado was most closely related to X. axonopodis genetic cluster 9.2. No other strain in this genetic cluster shared an identical sequence type. Avocado cv. Grand collet leaves from the youngest growth flush were infiltrated with a needleless syringe (10 inoculation sites per leaf and three replicates) with bacterial suspensions. Typical, water-soaked lesions that developed into black necrotic spots appeared 6 to 8 days after infiltration on all inoculated leaves when suspensions containing ~1 × 106 CFU ml-1 were used (i.e., ~7 × 102 CFU per inoculation site), while no lesions developed on leaves inoculated with Tris buffer or with suspensions containing ~1 × 104 CFU ml-1. One month after inoculation, mean Xanthomonas population sizes determined on KC semiselective medium (3) from ~1 cm2 leaf fragments showing black lesions ranged from 2 × 106 to 4 × 106 CFU per lesion, typical of a compatible interaction. A few colonies that recovered from lesions obtained after inoculation were typed by AFLP and were identical to the inoculated strain. An extensive branch and trunk canker of avocado caused by a Xanthomonas sp. has been reported in California (2). This bacterium did not cause lesions of avocado leaves or fruit after inoculation (2). This appears to be the sole previous report of a xanthomonad being pathogenic to avocado and the symptoms observed in the Seychelles appear therefore very different from the ones reported in California. No major outbreak of bacterial leaf spot of avocado has been reported in the Seychelles archipelago since 2003. References: (1) N. Ah-You et al. Int. J. Syst. Evol. Microbiol. 59:306, 2009. (2) D. A. Cooksey et al. Plant Dis. 77:95, 1993. (3) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (4) J. Rademaker et al. Phytopathology 95:1098, 2005.

Development of 14 minisatellite markers for the citrus canker bacterium, Xanthomonas citri pv. citri.

We screened the genome of Xanthomonas citri pv. citri strain 306 for tandem repeats. A multiplex polymerase chain reaction protocol was used to assess the genetic diversity of 239 strains of X. citri pv. citri from Asia. The total number of alleles per locus ranged from three to 20. Using pooled data sets, 223 different haplotypes were identified. Successful amplifications were obtained at most loci for seven other X. citri pathovars. This typing scheme is expected to be useful at different spatial scales for population studies of pathovars of X. citri, several of which cause plant diseases of economic importance.

First Report of Xanthomonas cucurbitae Causing Bacterial Leaf Spot of Pumpkin on Réunion Island.

Bacterial leaf spot of cucurbits caused by Xanthomonas cucurbitae (3) can be of economic importance in tropical and subtropical production areas, most often within the Cucumis, Cucurbita, and Citrullus genera. The bacterium induces angular, water-soaked leaf spots, which sometimes turn necrotic with a chlorotic halo. Scab-like lesions on fruit may also be observed (1). During 2000, water-soaked, angular leaf lesions were collected from pumpkin (Cucurbita pepo) in a production field located at Petit Serré, Réunion Island. Yellow-pigmented Xanthomonas-like bacterial colonies were isolated on yeast peptone glucose agar. Amplified fragment length polymorphism analysis was performed on four pumpkin isolates together with reference strains of X. cucurbitae (LMG 690 [type strain] and LMG 8663) and the type strain of all other valid Xanthomonas species using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (N. Ah-You, L. Gagnevin, P. A. D. Grimont, S. Brisse, X. Nesme, F. Chiroleu, L. Bui Thi Ngoc, E. Jouen, P. Lefeuvre, C. Verniére, and O. Pruvost, personal communication). The four isolates from pumpkin showed identical fingerprints and were most closely related to X. cucurbitae, with evolutionary genome divergences ≤0.05 (N. Ah-You et al., personal communication). One strain from diseased pumpkin (JW210-1) was further analyzed by multilocus sequence analysis using three housekeeping gene portions (atpD, dnaK, and gyrB) as described previously (N. Ah-You et al., personal communication). Although not fully identical, this strain displayed a similarity of >99% to the two reference strains of X. cucurbitae. Pumpkin and bottle-gourd (C. maxima), squash cv. aurore (C. pepo), cucumber cv. L-04 (Cucumis sativus), melon cv. cezanne (Cucumis melo), and watermelon cv. fou-nan (Citrullus lanatus) leaves were infiltrated (10 inoculation sites per leaf; three replicates) with bacterial suspensions prepared from strains JW210-1, LMG 690, and LMG 8663 and containing approximately 1 × 105 CFU ml-1. Negative controls consisted of leaves infiltrated with sterile tris buffer. Typical, water-soaked lesions that developed into necrotic spots were observed 6 to 8 days after inoculation for all inoculated plant species-strain combinations, but not for negative controls. One month after inoculation, mean Xanthomonas population sizes recovered from leaf lesions on KC semiselective medium (2) ranged from 1 × 107 to 1 × 108 CFU per lesion, typical of a compatible interaction. The reported outbreak was circumscribed to a single field and did not affect the local industry. No major outbreak of bacterial leaf spot of cucurbits has been reported on Réunion Island since 2000 on any host species of X. cucurbitae. References: (1) J. F. Bradbury. Page 309 in: Guide to Plant Pathogenic Bacteria. CAB International, Slough, UK, 1986. (2) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (3) L. Vauterin et al. Int. J. Syst. Bacteriol. 45:472, 1995.

Polymorphisms of 5,10-methylenetetralydrofolate reductase (MTHFR), fruit and vegetable intake, and the risk of stomach cancer.

Stomach cancer is a serious public health problem in China. 5,10-Methylenetetralydrofolate reductase (MTHFR) may be involved in both DNA methylation and DNA synthesis. Folate deficiency is associated with cancer risk that may be modulated by a genetic variation in the MTHFR gene in folate metabolism. The main goal of this study was to evaluate the association between polymorphisms of the MTHFR gene and the risk of stomach cancer. This study also explored the modification effects of fruit and vegetable intake (one of the main constituents is folate) on the risk of this disease. A population-based case-control study was conducted in Taixing, China, consisting of 206 newly diagnosed cases with primary stomach cancer and 415 healthy population controls. Polymorphisms of MTHFR C677T and A1298C were assayed by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) techniques. The data were analysed using the logistic regression model. No obvious association between the MTHFR A1298C polymorphism and the risk of stomach cancer was observed in this study. The frequencies of 677 C/C, C/T, and T/T were 34.5, 50.9, and 14.6%, respectively, in controls. The frequency of the MTHFR 677 wild homozygotic genotype was 25.8% in cases, which was lower than that in controls (34.5%). The adjusted odds ratio (OR) for the MTHFR 677 any T genotype was 2.05 (95% confidence interval (CI), 1.26-3.34) when compared with the C/C genotype. In the low fruit and vegetable intake group an increasing trend was observed with the T allele exposure, p = 0.0056. The adjusted ORs were 1.68 (95% CI = 0.86-3.29) for the C/T genotype and 3.58 (95% CI = 1.46-8.75) for the T/T genotype, respectively. The MTHFR 677 any T genotype was associated with an increased risk of primary stomach cancer among the Chinese population. Folate deficiency might modify the MTHFR gene polymorphism and influence the risk of stomach cancer.

Pathological Variations Within Xanthomonas campestris pv. mangiferaeindicae Support Its Separation Into Three Distinct Pathovars that Can Be Distinguished by Amplified Fragment Length Polymorphism.

ABSTRACT Bacterial black spot, caused by Xanthomonas campestris pv. mangiferaeindicae, is an important disease of mango (Mangifera indica). Several other plant genera of the family Anacardiaceae were described as host species for xanthomonads. We studied pathological variations among strains in a worldwide collection from several Anacardiaceae genera. Strains were classified into three pathogenicity groups. Group I strains (from the Old World) multiplied markedly in leaf tissue of mango and cashew (Anacardium occidentale). Group II strains (from Brazil) multiplied markedly in cashew leaf tissue, but not in mango. Moreover, mango leaves inoculated with group I and group II strains exhibited lesions with different morphologies, consistent with variations in symptomology previously reported on mango under field conditions. Group I strains produced black, raised lesions, consistent with the original description of the pathovar, whereas group II strains produced brownish, flat lesions. Group III strains produced a unique syndrome on ambarella (Spondias dulcis) and mombin (Spondias mombin). Based on evolutionary genome divergence derived from amplified fragment length polymorphism (AFLP) data, the three groups were genetically distinct and were related to groups 9.5, 9.6, and 9.4 of X. axonopodis identified by Rademaker, respectively. As each group was characterized by unique symptomology and/or host range, we propose that X. campestris pv. mangiferaeindicae be split into three pathovars of X. axonopodis: X. axonopodis pv. mangiferaeindicae, X. axonopodis pv. anacardii, and X. axonopodis pv. spondiae. Within pv. mangiferaeindicae sensu novo, AFLP data were consistent with that previously published for restriction fragment length polymorphism groups and suggested long-distance movement of the pathogen, likely through propagative material.

First Report in Myanmar of Xanthomonas axonopodis pv. mangiferaeindicae Causing Mango Bacterial Canker on Mangifera indica.

Bacterial canker of mango (or bacterial black spot) caused by Xanthomonas axonopodis pv. mangiferaeindicae (1) is a disease of economic importance in tropical and subtropical producing areas. X. axonopodis pv. mangiferaeindicae can cause severe infection in a wide range of mango cultivars and induces raised, angular, black leaf lesions, sometimes with a chlorotic halo. Several months after infection, leaf lesions dry and turn light brown or ash gray. Severe leaf infection may result in abscission. Fruit symptoms appear as small, water-soaked spots on the lenticels. These spots later become star shaped, erumpent, and exude an infectious gum. Often, a "tear stain" infection pattern is observed on the fruit. Severe fruit infections will cause premature fruit drop. Twig cankers are potential sources of inoculum and weaken branch resistance to winds (2). Suspected leaf lesions of bacterial canker were collected from mango nursery stock cv. Yin Kwe at a nursery in Yangon, Myanmar during March 2007. Nonpigmented Xanthomonas-like bacterial colonies were isolated on KC and NCTM3 semiselective agar media (4). Amplified fragment length polymorphism analysis was performed on three isolates from Myanmar and additional reference isolates of xanthomonads originating from Anacardiaceae (X. axonopodis pv. anacardii, X. axonopodis pv. mangiferaeindicae, X. axonopodis pv. spondiae, and X. translucens strains from pistachio) using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (1). On the basis of multidimensional scaling (1), the Myanmar isolates were identified as X. axonopodis pv. mangiferaeindicae and were most closely related to group B strains that were isolated from mango in India and Eastern Asia (2). Mango cv. Maison Rouge leaves, inoculated as previously reported (1) with the Myanmar isolates, showed typical symptoms of bacterial canker 1 week after inoculation. One month after inoculation, mean X. axonopodis pv. mangiferaeindicae population sizes ranging from 5 × 106 to 8 × 106 CFU per lesion were recovered from leaf lesions, typical of a compatible interaction (1). Mangifera indica L. probably evolved in the area that includes northwestern Myanmar (3) and to our knowledge, this is the first confirmed detection of X. axonopodis pv. mangiferaeindicae from Myanmar. Further surveys and strain collection will be necessary to evaluate its geographic distribution and prevalence in the country. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) L. Gagnevin and O. Pruvost. Plant Dis. 85:928, 2001. (3) S. K. Mukherjee. Page 1 in: The Mango, Botany, Production and Uses. R. E. Litz, ed. CAB International, Wallingford, UK, 1997. (4) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005.

Regulation of growth-regulated oncogene alpha expression by estrogen in human endothelial cells.

AIM: To study the effect of estrogen on expression of growth-regulated oncogene alpha (GROalpha) in human umbilical vein endothelial cells (HUVEC) in vitro. METHODS: Expressions of CXC chemokine GROalpha mRNA and protein were measured by Northern blotting assay and ELISA, respectively. The physiological significance of GROalpha expression was tested by static cell adhesion assay. RESULTS: Both the GROalpha mRNA and protein levels decreased markedly after HUVEC were exposured to 17beta-estradiol (2 0.05 micromol/L. Moreover, the inhibition of the protein was depended on the concentration of 17beta-estradiol. Tamoxifen (0.1 micromol/L), an estrogen receptor alpha antagonist, alone did not affect GROalpha protein expression, but can reverse the E2 -induced inhibition of GROalpha protein expression (by up to 50 %) and the binding of U937 cells to E2-treated HUVEC (by up to 40 %). CONCLUSION: Estrogen might functionally down-regulates GROalpha expression through estrogen receptor alpha on endothelial cells.

Abnormal liver function in workers exposed to low levels of ethylene dichloride and vinyl chloride monomer.

We investigated whether exposure to ethylene dichloride (EDC) and vinyl chloride monomer (VCM) resulted in increased risk of liver damage. Epidemiological information, including occupational, medical, smoking, and drinking history, was obtained by interview from 251 male workers. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyltransferase (GGT) were used as indicators of liver damage. Exposure to moderate or low levels of ECD and VCM resulted in a higher risk of developing abnormal ALT levels than did exposure to lower levels of the chemicals. Results were similar for AST. GGT was not associated with EDC or VCM exposure. Combined exposure to EDC and VCM showed a dose-response relationship in association with abnormal ALT levels. We concluded that relatively low concentrations of VCM and EDC cause liver damage.

Retrospective comparison of infusional 5-fluorouracil, doxorubicin, and mitomycin-C (modified FAM) combination chemotherapy versus palliative therapy in treatment of advanced gastric cancer.

About one-third of patients with gastric cancer are unresectable at the time of diagnosis. Their median survival is < 6 months, with a grave prognosis. The purpose of this study was to assess the efficacy of a modified FAM (mFAM) regimen in advanced gastric cancer. We retrospectively reviewed the clinical records of 409 advanced gastric cancer patients who had not received curative surgery. Among 409 patients, 202 patients were treated with an mFAM regimen (infusional 5-FU + doxorubocin + mitomycin-C), and 207 patients received no chemotherapy (control group). No differences were found in clinical parameters between the two groups. The 1-year survival rates were 34.1% for the mFAM-treated group and 22.5% for the control group (p = 0.0135). In subset analysis, a higher 1-year survival rate was demonstrated in patients with mFAM and palliative surgery. Of the 154 evaluable patients in the mFAM-treated group, the response rate was 17.5%. In these patients, median response duration was 30 weeks, and progression-free survival was 23 weeks. Overall toxicity of mFAM regimen was relatively tolerable and reversible. In conclusion, FAM combination chemotherapy, which has been used as a standard therapy, prolonged survival after modification of the administration schedule and combination with palliative surgery. A prospective randomized study is warranted to confirm this conclusion from our retrospective study.

Correlation between K-ras gene mutation and prognosis of patients with nonsmall cell lung carcinoma.

BACKGROUND: Mutations at codons 12, 13, and 61 of the three ras genes, H-ras, K-ras, and N-ras, convert these genes into active oncogenes. It appears that ras gene mutations can be found in a variety of tumor types. The purpose of this study was to evaluate the clinical significance of K-ras gene mutation in nonsmall cell lung carcinoma (NSCLC). METHODS: The authors analyzed 58 NSCLC patients for mutations at codons 12, 13, and 61 of the K-ras gene and correlated the findings with the tumor stage and patient survival. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and the direct nucleotide sequencing method were used to detect mutations after amplification of ras specific sequences by PCR. RESULTS: Fourteen mutations (24%) of ras genes were found, all at codon 12 of the K-ras gene. GGT to GAT transition was the predominant mutational pattern. There was a significant association between K-ras mutation and the tumor stage (i.e., the higher the stage, the higher the mutation rate) (P = 0.014). Using univariate analysis, the presence of K-ras mutation in paraffin embedded tissue from patients who received treatment with curative intent was associated with a shorter survival (P = 0.039). The median survival duration for patients with or without K-ras mutation was 9 and 30 months, respectively. The Cox proportional hazards model also predicted a higher risk for patients with K-ras mutations (P = 0.047). CONCLUSIONS: K-ras mutations, present in a subset of NSCLC, are associated with tumor progression and shortened patient survival.