Whole-Genome Sequencing and Comparative Analysis of Yersinia pestis, the Causative Agent of a Plague Outbreak in Northern Peru.

The plague is a zoonotic disease caused by the bacterium Yersinia pestis. Here, we report the complete genome sequence of the Y. pestis strain INS, which was isolated from swollen lymph gland aspirate (bubo aspirate) of an infected patient from a pneumonic outbreak in 2010 in northern Peru.

Whole Genome Sequencing and Comparative Analysis of Bartonella bacilliformis Strain INS, the Causative Agent of Carrion's Disease.

Bartonella bacilliformis is the etiological agent of human bartonellosis, which is highly endemic to Peru. Here, we report the first genome that was sequenced and analyzed from an isolate of B. bacilliformis strain INS, which originally was isolated from the blood of an infected patient with an acute phase of Carrion's disease from Jaén-Cajamarca, Peru.

First Report of "Candidatus Liberibacter solanacearum" on Tomato in El Salvador.

In April of 2012, tomato plants (Solanum lycopersicum) grown near the town of Yuroconte in the municipality of La Palma, Chalatenango, El Salvador, were observed with symptoms resembling those of "Candidatus Liberibacter solanacearum" infection. The symptoms included overall chlorosis, severe stunting, leaf cupping, excessive branching of axillary shoots, and leaf purpling and scorching (1,2,3). Disease incidence in several fields in the area ranged from 40 to 60%. Heavy infestations of the potato/tomato psyllid, Bactericera cockerelli, were observed in the affected fields and this insect has been shown to transmit "Ca. L. solanacearum" to tomato and other solanaceous species (1,2,3). Leaf samples and psyllids were collected from one of the fields and total DNA was purified from the leaves of 8 and 10 symptomatic and asymptomatic plants, respectively (2,3). DNA was also extracted from the psyllids and the samples were tested by PCR for species confirmation. PCR oligonucleotide primers specific for both 16S rDNA (OA2 and OI2c) and a gene for a surface antigen for the outer membrane protein (OMB) (OMB 1482f and 2086r) of "Ca. L. solanacearum" were used to confirm the presence of the bacterium in infected tomatoes (1). Four of the eight symptomatic tomatoes (50%) tested positive for "Ca. L. solanacearum" using both primer pairs and all asymptomatic plants were negative for the bacterium. The collected psyllids were first identified through a morphological key, then verified using species-specific PCR primers (CO1 F3 and CO1 meltR) that generated a 94-bp fragment that was consistent with DNA from B. cockerelli (4). Amplicons generated with DNA from two plant samples with each primer pair were cloned and four clones of each of the four amplicons were sequenced. BLASTn analysis of the 16S rDNA consensus sequences from the clones (1,168 bp; deposited in GenBank as Accession Nos. KC768318 and KC768319) showed 100% identity to "Ca. L. solanacearum" sequences in GenBank (HM246509 and HM245242, respectively). Two OMB consensus sequences were 98% identical (deposited in GenBank as KC768326 and KC768327) and both sequences were 97 to 100% identical to a number of "Ca. L. solanacearum" sequences in GenBank (e.g., CP002371, FJ914617, JN848754, and JN848752). To our knowledge, this is the first report of "Ca. L. solanacearum" associated with tomato in El Salvador and the first formal report of the bacterium in the country. This bacterium has caused millions of dollars in losses to the tomato industry in New Zealand, Mexico and the United States (2,3). Tomatoes are an economically important commodity in Central America and are severely damaged by "Ca. L. solanacearum" infection. The confirmation of "Ca. L. solanacearum" infections in El Salvador alerts the agricultural sector to the presence of this serious pathogen. References: (1) J. M. Crosslin. Southwest. Entomol. 36:125, 2011. (2) L. W. Liefting et al. Plant Dis. 93:208, 2009. (3) J. E. Munyaneza et al. Plant Dis. 93:1076, 2009. (4) K. D. Swisher et al. Environ. Entomol. 41:1019, 2012.

Efecto de un anticonceptivo oral con drospirenona/etinilestradiol (3 mg - 30 mg) sobre el hiperandrogenismo clínico y bioquímico en pacientes con síndrome de ovario poliquístico / Effect of an oral contraceptive containing drospirenone / ethinyl estradiol (3 mg - 30 mg) on the clinical and biochemical hyperandrogenism in patients with polycystic ovary syndrome

El síndrome de ovario poliquístico es un desorden heterogéneo de etiología incierta, que cual afecta entre el 6% y 10% de las mujeres en edad reproductiva. Una de las opciones terapéuticas especificas es el uso de los anticonceptivos orales, con la progestina, drospirerona, la cual, es un análogo de la espironolactona que posee actividad antimineralcorticoides y antiandrogénica. El objetivo de este estudio fue determinar el efecto del anticonceptivo oral combinado (EE: 30 MG y DRP: 3 mg) en el perfil bioquímico y clínico en una población de mujeres venezolanas con síndrome de ovario poliquístico. De las 20 pacientes incluidas en el estudio, 18 completaron satisfactoriamente el estudio, con una buena tolerancia al tratamiento. Se observó una disminución del IMC de 23,94 en condición basal a 23,73 kg/m². Los niveles de andrógenos se encontraron disminuidos significativamente en comparación a la basal; testosterona total cayó de 1,4 ng/mL a 0,67 ng/mL; Testosterona libre bajo de 3 pg/mL a 1,38 pg/mL; DHEAS disminuyó de 1,65 µg/mL a 1,08 µg/mL y androstenediona de 2,50 ng/mL a 1,55 ng/mL. En conclusión nuestros resultados reportan que el uso de un anticonceptivo oral que contiene 30 mg de EE y 3 mg de progestina, drospirerona en una población de mujeres venezolanas con síndrome de ovario poliquístico condujo a una disminución de los niveles de andrógenos al mismo tiempo que se evidenció un incremento de la SHBG, así como una reducción no significativa del peso corporal de este grupo de pacientes y una mejoría clínica del hirsutismo.

Policystic ovarian syndrome is a heterogeneous disorder wich etiology remained uncertain and affects 6%-10% of reproductive age women. Most recommended therapy is oral contraceptives with progestins. Drospirenone is an espironolactone analogue exhibits a partial antiandrogenic action and has predominant anti-mineralocorticoid properties. This is a prospective trial to determine efficacy of a drospirenone-containing combined oral contraceptives in venezuelan women with polycystic ovary-syndrome. Twenty women were conducted into this trial, although 18 were treated. With treatment, BMI fell by 0,21 kg/m(2) in the study group. During therapy, the levels of testosterone, free testosterone, Delta (4)-androstenedione, and androstenedione decreased significantly, whereas sex hormone-binding globulin increased significantly. Treatment of women with polycystic ovary-syndrome with drospironene containing combined oral contraceptives formulations is effective in decreasing hirsutism, androgen levels and BMI.

Two different profiles of peach allergy in the north of Spain.

BACKGROUND: Peach allergy has two different patterns: central Europe with oral allergy syndrome (OAS) related to a primary sensitization to birch pollen Bet v 1 and profilins and southern Europe with mostly systemic symptoms, in many cases due to sensitization to lipid-transfer proteins. METHODS: Thirty peach-allergic patients with positive skin and food challenge tests and 29 control subjects were included. Skin prick tests (SPT) with inhalant allergens, commercial peach and apple extracts and native Pru p 3 were performed. In vitro specific immunoglobulin (Ig) E to grass pollen, birch pollen, peach, apple, rBet v 1, rBet v 2 and rPhl p 12 was determined by CAP, and rBet v 1, rMal d 1, rMal d 4, rMal d 3 and rPru p 3 using the ADVIA-Centaur platform. Basophil activation test (BAT) with commercial peach extract, commercial apple extract, nPru p 3, rMal d 3, rMal d 1 and rMal d 4 was also performed. RESULTS: Pru p 3 was the major allergen in the patient group from northern Spain. Sensitization to this allergen was found in 100% of the patients with systemic symptoms or contact urticaria. Only 60% of OAS patients were sensitized to Pru p 3, being all of them sensitized to profilins and 60% of them to allergens of the Bet v 1 family. Specific IgE determination and BAT using recombinant allergens (rPru p 3) show specificity and sensitivity values close to 100%. CONCLUSIONS: Most peach-allergic patients coming from the north of Spain present systemic symptoms after ingestion of peach, Pru p 3 being the main allergen. Patients with OAS present profilin-Bet v 1-related sensitization. Thus, in the north of Spain our patients show a mixed central-south Europe pattern with LTP-profilin-Bet v 1 sensitization depending on the symptoms presented. The use of natural and recombinant plant allergens, allows establishing the sensitization patterns to the different allergens studied.

Influence of the histidine tail on the structure and activity of recombinant chlorocatechol 1,2-dioxygenase.

We present two efficient expression systems for the chlorocatechol 1, 2-dioxygenase (CCD) from Pseudomonas putida. In the first, CCD (encoded by the clcA gene) was expressed in the pETCLCA vector with the addition of an N-terminal histidine tail. After purification, the enzyme (CCD 6xHis) was proteolytically cleaved with thrombin to remove the His tail. The CD spectra of the cleaved and uncleaved enzymes present only minor differences, indicative of correct protein folding. However, the activity of CCD 6xHis, over a wide range of pH, was typically five times lower. This may be the result of steric hindrance caused by the histidine tail. These data are consistent with results obtained using an alternative construct employing a vector which produces a protein product devoid of the His tail. These results suggest that the His tail may induce subtle effects close to the active site which compromise the recovery of full biological activity.

Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4.

Growth of Alcaligenes eutrophus JMP134 on 2,4-dichlorophenoxyacetate requires a 2,4-dichlorphenol hydroxylase encoded by gene tfdB. Catabolism of either 2,4-dichlorophenoxyacetate or 3-chlorobenzoate involves enzymes encoded by the chlorocatechol oxidative operon consisting of tfdCDEF, which converts 3-chloro- and 3,5-dichlorocatechol to maleylacetate and chloromaleylacetate, respectively. Transposon mutagenesis has localized tfdB and tfdCDEF to EcoRI fragment B of plasmid pJP4 (R. H. Don, A. J. Wieghtman, H.-J. Knackmuss, and K. N. Timmis, J. Bacteriol. 161:85-90, 1985). We present the complete nucleotide sequence of tfdB and tfdCDEF contained within a 7,954-base-pair HindIII-SstI fragment from EcoRI fragment B. Sequence and expression analysis of tfdB in Escherichia coli suggested that 2,4-dichlorophenol hydroxylase consists of a single subunit of 65 kilodaltons. The amino acid sequences of proteins encoded by tfdD and tfdE were found to be 63 and 53% identical to those of functionally similar enzymes encoded by clcB and clcD, respectively, from plasmid pAC27 of Pseudomonas putida. P. putida(pAC27) can utilize 3-chlorocatechol but not dichlorinated catechols. A region of DNA adjacent to clcD in pAC27 was found to be 47% identical in amino acid sequence to tfdF, a gene important in catabolizing dichlorocatechols. The region in pAC27 does not appear to encode a protein, suggesting that the absence of a functional trans-chlorodienelactone isomerase may prevent P. putida(pAC27) from utilizing 3,5-dichlorocatechol.