Antiviral activity of maca (Lepidium meyenii) against human influenza virus

Objective: To investigate antiviral activity of maca to reduce viral load in kidney (MDCK) cells infected with influenza type A and B viruses (Flu-A and MFalud-inB-, Dreasrpbeyc ctiavneilny)e. Methods: Maca were extracted with methanol (1:2, v/v). The cell viability and toxicity of the eaxgtariancstts Fwluer-eA e avnaldu aFtleud- oBn v MirDusCeKs cwealsls a usssianyge dm uetshinogd aM TteTs ta sfosar yd. eAtenrtmiviinrainl ga ctthiev itiyn hoifb citoimonp oouf nthdes cytopathic effect on cell culture and multiplex RT-PCR. Results: The methanol extract of maca showed low cytotoxicity and inhibited influenza-induced cytopathic effect significantly, while viral load was reduced via inhibition of viral growth in MDCK infected cells. Maca contains potent inhibitors of Flu-A and Flu-B with a selectivity index [cytotoxic concentration 50%/IC50] of 157.4 and 110.5, respectively. Conclusions: In vitro assays demonstrated that maca has antiviral activity not only against Flu-A (like most antiviral agents) but also Flu-B viruses, providing remarkable therapeutic benefits.Financial support of this study was provided by AECID grants (PCI: C/033641/10) and AGAUR (MAT2009-11503, MAT2012-36205, 2009SGR-1208). JDVM support was provided by 1st Concurso Incentivo a la Investigación de la Universidad Peruana de Ciencias Aplicadas, Lima, Peru.Revisión por pare

Carrion's Disease: More Than a Sand Fly-Vectored Illness

Carrion’s disease is a biphasic illness (S1 Fig) caused by an infection of Bartonella bacilliformis, a bacterium that is transmitted through bites of certain phlebotomine sand flies in the Andean valleys of Peru and in some areas of Ecuador and southern Colombia [1,2]. The acute phase, called Oroya fever, is a serious, life-threating illness that mainly affects immunologically naïve populations, such as children. It is also of special concern in pregnant women, because high mortality rates have been described as well as miscarriages, preterm births, and fetal deaths [3]. In this acute phase, the absence or delay of antibiotic treatment may lead to fatal outcomes. In fact, it is considered that, in the pre-antibiotic era, the lethality of this illness ranked between 40% and 88% [1,2]. In the chronic phase, classically considered to occur in previously exposed inhabitants, B. bacilliformis induce endothelial cell proliferation, producing skin lesions called Peruvian warts. In this phase, the lethality is very low [1]. Additionally, the presence of asymptomatic carriers is frequent, although the real numbers remain uncertain because of the difficulty in detecting these subjects

Carrion's disease: an eradicable illness?

Carrion's disease is a neglected tropical disease caused by Bartonella bacilliformis, a vector-borne pathogen restricted to the Andean valleys of Peru, Ecuador and Colombia. Carrion's disease is a biphasic illness; in the acute phase the case-fatality rate can be as high as 88 %, related to high parasitemia, arriving to almost all erythrocytes, and secondary bacterial infections close related with the development of transient immunosuppression in the earlier illness phases. In addition, there are an undefined number of asymptomatic carriers that are reservoirs of the etiological agent of Carrion's disease in endemic areas, they make take into account due to they are the perpetuators of this disease. The actual scenario of Carrion's disease, in which the illness is arriving to new areas, due to the expansion of the vector's distribution, suggests that now may be a crucial time to design a strategy focusing on its elimination

Multi-Locus Sequence Typing of Bartonella bacilliformis DNA Performed Directly from Blood of Patients with Oroya's Fever During a Peruvian Outbreak.

Background Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected tropical poverty-linked illness. This infection is endemic of Andean regions and it is estimated that approximately 1.7 million of South Americans are at risk. This bacterium is a fastidious slow growing microorganism, which is difficult and cumbersome to isolate from clinical sources, thereby hindering the availability of phylogenetic relationship of clinical samples. The aim of this study was to perform Multi Locus Sequence Typing of B. bacilliformis directly in blood from patients diagnosed with Oroya fever during an outbreak in Northern Peru. Methodology/Principal Findings DNA extracted among blood samples from patients diagnosed with Oroya’s fever were analyzed with MLST, with the amplification of 7 genetic loci (ftsZ, flaA, ribC, rnpB, rpoB, bvrR and groEL) and a phylogenetic analysis of the different Sequence Types (ST) was performed. A total of 4 different ST were identified. The most frequently found was ST1 present in 66% of samples. Additionally, two samples presented a new allelic profile, belonging to new STs (ST 9 and ST 10), which were closely related to ST1. Conclusions/Significance The present data demonstrate that B. bacilliformis MLST studies may be possible directly from blood samples, being a promising approach for epidemiological studies. During the outbreak the STs of B. bacilliformis were found to be heterogeneous, albeit closely related, probably reflecting the evolution from a common ancestor colonizing the area. Additional studies including new samples and areas are needed, in order to obtain better knowledge of phylogenetic scenario B. bacilliformisThis work has been supported by the Spanish Network for the Research in Infectious Diseases [REIPI RD12/0015],by Generalitat de Catalunya, Departament d’Universitats, Recerca i Societat de la Informació [2014 SGR 26] and by by a grant of the Instituto de Salud Carlos III - Spain (PI11/ 00983) which included FEDER funds (JR). MJP has a postdoctoral fellowship from CONCYTEC/ FONDECYT (grant number: CG05-2013- FONDECYT). JR has a fellowship from the programRevisión por pare

Etiological and molecular diagnostic of Carrion’s disease in patients from Cajamarca - Perú

[EN] Poster presented in the poster session in the 15th ICID Abstracts. June 13th-16th 2012, Bangkok, Thailand. Session: Emerging Infectious Diseases. Date: Friday, June 15, 2012. Room: Poster & Exhibition Area.Background: Bartonellagenus is a group of facultative intracellular pathogens that posses able to survive and proliferate inside of erythrocytes. Classified within this genus,Bartonella bacilliformisis of special relevance. This microorganism is the etiological agent of the so called Carrion’s Disease (Human bartonellosis). Additionally the presence of sub-clinical cases (asymptomatyc carriers) is of special interest, because acts as a reservoir of this illness. Carrión’s Disease is an endemic illnes in Perú, affecting in a special manner the north interandean valleys. However, the current in use diagnostic techniques (Giemsa Stain) possess low sensitivity and specificity, and due to the fact thatB. bacilliformispossess a low growth (weeks), bacterial cultures lacks of clinical utility. Thus suspictious cases frequently are not confirmed, and the real relevance of this illness remains underestimated. This work is addressed to the direct identification from blood samples ofBartonella baciliformisusing a conventional PCR. All patients were from the Cajamarca area being enrolled by the Epidemiological Surveillance program of DIRESA. Methods: The samples were processed at arriving to the laboratory, by molecular and microbiological techniques. Thus samples were cultured in Blood Columbia Agar (10%), in anaerobic conditions at 28 ◦C for a period of 2 months. Positive cultures were both Giemsa stained and identified by the amplification of a fragment the 16S rRNA gene. Genetic material was directly extracted from blood samples using the Kit High Pure (Roche diagnostic), and a fragment of 438 bp of the 16S rRNA gene was amplified withBartonellagenus specific primers. All positive PCR were sequenced (Macrogen-Korea). Results: A total of 134 blood samples were processed, from this 12 (8.9%) grown in blood agar, while in 18 (13.4%), including the aforementioned 12, the 16 s rRNA gene was amplified. In all cases the sequence analysis showed the presence ofB. bacilliformis Conclusion: Although microbiological culture is the gold standard in the identification ofBartonellaspp., this technique possess strong limitations due to the low growth of these microorganisms. However, the PCR is a rapid technique, possessing a high sensibility and specificity that may be used as routine diagnostic tool for the identification of Carrion’s Disease.Revisión por pare

Carrion's disease after blood transfusion

Bartonella bacilliformis is a pathogen that is endemic in some areas of the Andean region of Peru, southern Ecuador and southern Colombia. This pathogen causes so-called Carrion's disease, a biphasic disease with acute and chronic phases (called Oroya fever and "Peruvian wart" respectively). In the absence or delay of antibiotic treatment, the mortality rate in the acute phase is up to 88%1. The acute phase is characterised by fever and severe anaemia and may be followed, several weeks or months later, by the chronic eruptive phase due to endothelial cell proliferation2. No animal reservoir has been identified to date and it is considered that healthy carriers act as a pathogen reservoir in endemic areas

Removal of Paracetamol Using Effective Advanced Oxidation Processes

Fenton, photo-Fenton, and photo-induced oxidation, were investigated and compared for the treatment of 0.26 mmol L-1 of paracetamol (PCT) in a deionised water matrix, during a reaction span of 120.0 min. Low and high Fenton reagent loads were studied. Particularly, the initial concentration of Fe2+ was varied between 0.09 and 0.18 mmol L-1 while the initial concentration of H2O2 was varied between 2.78 and 11.12 mmol L-1. The quantitative performance of these treatments was evaluated by: (i) measuring PCT concentration; (ii) measuring and modelling TOC conversion, as a means characterizing sample mineralization; and (iii) measuring cytotoxicity to assess the safe application of each treatment. In all cases, organic matter mineralization was always partial, but PCT concentration fell below the detection limit within 2.5 and 20.0 min. The adopted semi-empirical model revealed that photo induced oxidation is the only treatment attaining total organic matter mineralization (¿MAX= 100% in 200.0 min) at the expense of the lowest kinetic constant (k = 0.007 min-1). Conversely, photo-Fenton treatment using high Fenton reagent loads gave a compromise solution (¿MAX= 73% and k = 0.032 min-1). Finally, cytotoxicity assays proved the safe application of photo-induced oxidation and of photo-Fenton treatments using high concentrations of Fenton reagents.Peer ReviewedPostprint (published version

Identification of human papillomavirus as a preventive strategy for cervical cancer in asymptomatic women in the Peruvian Andes

Objective: To detect the most prevalent human papillomavirus (HPV) genotypes samples of asymptomatic Peruvian women by analyzing the correlation betwe ienn c Pearvpiacnailc somlaeoaur (PAP)-stained cervical tests and PCR-sequencing. Methods: A total of 254 women attending routine gynecological examinations were included in pthaitsh ostluogdiys.t Tahned scalmaspsliefise dw ebrye tahnea Blyeztehde sbdya PsAysPt etmec.h HnPiqVu ea manpdli feicxaatmioinn ewda su nddoenre au msinicgr othsceo pprei mbeyr as specific for E1 region and positive specimens were confirmed by direct sequencing. Results: The prevalence of HPV was investigated in 254 cervical scrape samples by PCR. PAP smear showed that 94.9% cases had normal morphology and 5.1% had an inflammatory pattern; 2p0r.e5v%a lwenert eg efonuontydp teo ibne c ionrfreeclatetido nw iwthit hH PchVa, ncgoems pinri scienrgv i2c0a dl icfyfetorelongt yg.enotypes. HPV16 was the most Conclusions: Our results suggest the HPV is very frequent even in women with negative PAP, eannddo PceCrRvi csaele smasm tpol ebs.e Itdheen tbifeicsat toiopnt ioofn t htoe HdePtVe rgmeinnoety tphee inc aaussyamtipvteo magateinc tw oofm HePnV m ianyf eaclltoiown t hine nimatpulreaml henisttaotriyo no fo tfh ea pdpisroepasreia aten dp rthope hsyulbascetqicu emnet adseuvreelso pwmheicnht omf acye rhviacvael ma adliirgencatn [email protected] work has been partially supported by Universidad Peruana de Ciencias Aplicadas (UPC), Instituto de Investigación Nutricional and Instituto de Investigación de Efrnomfe rtmheed pardoegsr aImnf ecciosas, Lima, Peru. JR has a fellowship I3, of the ISCIII (Grant No. CES11/012), and LJDV from the Generalitat de Catalunya (2009SGR1208).Revisión por pare

Development and characterisation of highly antibiotic resistant Bartonella bacilliformis mutants

The objective was to develop and characterise in vitro Bartonella bacilliformis antibiotic resistant mutants. Three B. bacilliformis strains were plated 35 or 40 times with azithromycin, chloramphenicol, ciprofloxacin or rifampicin discs. Resistance-stability was assessed performing 5 serial passages without antibiotic pressure. MICs were determined with/without Phe-Arg-beta-Napthylamide and artesunate. Target alterations were screened in the 23S rRNA, rplD, rplV, gyrA, gyrB, parC, parE and rpoB genes. Chloramphenicol and ciprofloxacin resistance were the most difficult and easiest (>37.3 and 10.6 passages) to be selected, respectively. All mutants but one selected with chloramphenicol achieved high resistance levels. All rifampicin, one azithromycin and one ciprofloxacin mutants did not totally revert when cultured without antibiotic pressure. Azithromycin resistance was related to L4 substitutions Gln-66 --> Lys or Gly-70 --> Arg; L4 deletion Delta62-65 (Lys-Met-Tyr-Lys) or L22 insertion 83::Val-Ser-Glu-Ala-His-Val-Gly-Lys-Ser; in two chloramphenicol-resistant mutants the 23S rRNA mutation G2372A was detected. GyrA Ala-91 --> Val and Asp-95 --> Gly and GyrB Glu474 --> Lys were detected in ciprofloxacin-resistant mutants. RpoB substitutions Gln-527 --> Arg, His-540 --> Tyr and Ser-545 --> Phe plus Ser-588 --> Tyr were detected in rifampicin-resistant mutants. In 5 mutants the effect of efflux pumps on resistance was observed. Antibiotic resistance was mainly related to target mutations and overexpression of efflux pumps, which might underlie microbiological failures during treatments