Spatial, temporal, and motivational changes due to the COVID-19 pandemic in a nature-based leisure activity - A global survey of birders.

Birdwatchers contribute an immense amount of data to citizen science databases. Thus, birding is important from the leisure perspective and from nature conservation. During the COVID-19 pandemic, we studied birdwatchers on a global scale in over 50 countries by applying the model of behavior change, which focuses on changes in opportunity (spatial, temporal), motivation, and capability (avoidance behavior). The sample consisted of 5051 participants (3437 men, 1575 women, mean age 49 years). Birders changed their spatial behavior to more local birding and to avoidance behavior by choosing different places and different clock times. Concerning motivation, being outdoors showed the highest increase and being with friends the strongest decrease. Higher specialized birders experienced a stronger shift toward regional birding. Birders that focused on new, local, or unrewarding places experienced an increase in motivation. Our study empirically supports the behavior change model and highlights the need to address the heterogeneity of the recreationists.

Cattle Farming and Plantation Forest are Associated with Bartonella Occurrence in Wild Rodents.

Bartonella spp. are intracellular hemotropic bacteria primarily transmitted by arthropod vectors to various mammalian hosts, including humans. In this study, we conducted a survey on wild populations of sigmodontine rodents, Akodon azarae and Oxymycterus rufus, inhabiting the Paraná River delta region. The study involved eight grids organized in a crossed 2 × 2 design, where four of the grids were exposed to cattle while the other four were not, and four grids were located in implanted forest while the remaining four were in natural grasslands. Our objective was to examine whether the occurrence of Bartonella spp. in rodents was associated with silvopastoral activities (cattle raising associated with timber production) conducted in the region. Additionally, we evaluated the associations between Bartonella infection and other environmental and host factors. We present compelling evidence of a significant positive association between Bartonella prevalence and the presence of implanted forests and cattle. Furthermore, we identified the presence of a Bartonella genotype related to the pathogen Bartonella rochalimaea, infecting both A. azarae and Ox. rufus. These findings suggest that anthropogenic land-use changes, particularly the development of silvopastoral practices in the region, may disrupt the dynamics of Bartonella.

Rickettsia spp., Ehrlichia sp. and Candidatus Midichloria sp. associated to ticks from a protected urban area in Buenos Aires City (Argentina).

The aim of this study was to determine the infection with Rickettsiales in ticks and birds from the main protected urban area of Buenos Aires City (Argentina). One Amblyomma aureolatum (0.2%) and one Ixodes auritulus (0.1%) were positive by PCR targeting Rickettsia 23S-5S rRNA intergenic spacer. Phylogenetic analysis shows to findings in A. aureolatum are closely to Rickettsia bellii and for I. auritulus are related to 'Candidatus Rickettsia mendelii'. One I. auritulus (0.1%) and three A. aureolatum (0.6%) were positive by PCR for a fragment of the 16S rRNA gene of the Anaplasmataceae family. The sequences obtained from A. aureolatum were phylogenetically related to Midichloriaceae endosymbionts. The sequence from I. auritulus s.l. had 100% identity with Ehrlichia sp. Magellanica from Chile and two genotypes of Ehrlichia sp. from Uruguay. The results of our study show that Rickettsia and Ehrlichia are present in ticks in the main protected urban area of Buenos Aires City.

Evolving determinants of carotid atherosclerosis vulnerability in asymptomatic patients from the MAGNETIC observational study.

MRI can assess plaque composition and has demonstrated an association between some atherosclerotic risk factors (RF) and markers of plaque vulnerability in naive patients. We aimed at investigating this association in medically treated asymptomatic patients. This is a cross-sectional interim analysis (August 2013-September 2016) of a single center prospective study on carotid plaque vulnerability (MAGNETIC study). We recruited patients with asymptomatic carotid atherosclerosis (US stenosis > 30%, ECST criteria), receiving medical treatments at a tertiary cardiac rehabilitation. Atherosclerotic burden and plaque composition were quantified with 3.0 T MRI. The association between baseline characteristics and extent of lipid-rich necrotic core (LRNC), fibrous cap (CAP) and intraplaque hemorrhage (IPH) was studied with multiple regression analysis. We enrolled 260 patients (198 male, 76%) with median age of 71-y (interquartile range: 65-76). Patients were on antiplatelet therapy, ACE-inhibitors/angiotensin receptor blockers and statins (196-229, 75-88%). Median LDL-cholesterol was 78 mg/dl (59-106), blood pressure 130/70 mmHg (111-140/65-80), glycosylated hemoglobin 46 mmol/mol (39-51) and BMI 25 kg/m2 (23-28); moreover, 125 out of 187 (67%) patients were ex-smokers. Multivariate analysis of a data-set of 487 (94%) carotid arteries showed that a history of hypercholesterolemia, diabetes, hypertension or smoking did not correlate with LRNC, CAP or IPH. Conversely, maximum stenosis was the strongest independent predictor of LRNC, CAP and IPH (p < 0.001). MRI assessment of plaque composition in patients on treatment for asymptomatic carotid atherosclerosis shows no correlation between plaque vulnerability and the most well-controlled modifiable RF. Conversely, maximum stenosis exhibits a strong correlation with vulnerable features despite treatment.

A novel Ehrlichia strain (Rickettsiales: Anaplasmataceae) detected in Amblyomma triste (Acari: Ixodidae), a tick species of public health importance in the Southern Cone of America.

The aim of this work was to report the detection of a putative novel Ehrlichia strain associated with the tick Amblyomma triste. Free-living adult ticks determined as A. triste were collected by drag-sampling in Argentina and Uruguay. Molecular detection of Ehrlichia agents was performed targeting three different loci: 16S rRNA gene, dsb gene and a fragment of groESL heat shock operon. In total, 164 adults of A. triste (38 from INTA E.E.A Delta del Paraná in Argentina and 126 from Toledo Chico in Uruguay) were analyzed. One tick (0.6%) collected in INTA E.E.A. Delta del Paraná (Argentina) was positive. The phylogenetic analyses show that the Ehrlichia strain found in this study (named Ehrlichia sp. strain Delta) represents an independent lineage within the genus Ehrlichia, close to E. chaffeensis and E. muris. This is also the first report of an Ehrlichia agent infecting the tick A. triste. The medical and veterinary significance of Ehrlichia sp. strain Delta remains to be demonstrated. However, it is important to mention that adults of A. triste are aggressive to humans and domestic mammals. Therefore, the potential role of A. triste in the transmission of Ehrlichia agents to humans or domestic animals across its distributional range should be highlighted, even more considering that Ehrlichia sp. strain Delta is phylogenetically related to the zoonotic E. chaffeensis, which is recognized as pathogenic to both humans and animals.

Bartonella spp. associated with rodents in an urban protected area, Buenos Aires (Argentina).

At least 15 of the 30 Bartonella species are involved in human pathologies, and several of them are associated with rodents and their fleas. The aims of this study were detect and molecularly characterize the Bartonella infections in rodents from an urban protected area of ​Buenos Aires City (Argentina). A total of 186 rodents were captured and identified. For PCR of the 16S rRNA fragment, 23.7 % of the samples tested positive, and two groups (GrA and GrB) were identified. Likewise, the comparison between the sequences obtained for the gltA gene determined the presence of three genotypes, closely related to Bartonella spp. detected in sigmodontine rodents and their fleas in the Americas, which form a well-separated clade. The high prevalence of Bartonella in rodents from an urban protected area of ​Buenos Aires city is relevant from a public health perspective.

Molecular detection and identification of Bartonella in the cat flea Ctenocephalides felis felis collected from companion animals in a border area in northeastern Argentina.

Molecular methods were used to detect and identify Bartonella species in the cat fleas Ctenocephalides felis felis from Puerto Iguazú, a border area in northeastern Argentina. The fleas were collected from 12 household animals, 9 dogs (Canis lupus familiaris) and 3 cats (Felis silvestris catus) during July 2016. Out of 15C. f. felis analyzed for PCR, only one flea collected from a cat was positive (6.66%) in screened for Bartonella spp. based on the gltA gene. Bartonella clarridgeiae was identified in the genetic analyses, this specimen clustered monophyletically with others B. clarridgeiae isolated from different geographical origins (1.0 PP), even, all shared the same haplotype. The results obtained provide evidence of the presence of B. clarridgeiae in cat fleas from Argentina suggesting the probable presence of related flea-borne diseases in the region and the role of cat fleas in the transmission of Bartonella among mammals including humans.

Borrelia spp. in ticks and birds from a protected urban area in Buenos Aires city, Argentina.

This study was aimed to know epidemiological aspects of Borrelia spp. in a protected urban area of Buenos Aires city, Argentina, where thousands of people visit this area for recreational purposes. Ticks were collected from vegetation, birds and dogs. Three hundred and forty birds belonging to 43 species, 41 genera, 18 families and six orders were captured (90.3% corresponded to the order Passeriformes). One hundred and twenty ticks were collected from 47 birds (13.8%) belonging to 10 species (23.2%), all of them from to the order Passeriformes (Emberizidae, Furnariidae, Parulidae, Thraupidae, Troglodytidae, Turdidae). Ticks were identified as Ixodes auritulus (56 larvae, 44 nymphs and 8 females) and Amblyomma aureolatum (1 larva and 11 nymphs). One thousand and ninety-one ticks collected from vegetation, 100 ticks collected from birds, and 89 ticks from dogs were tested for Borrelia infection by PCR trials targeting the flagellin (fla) and 16S rRNA genes. In addition, 101 blood and 168 tissue samples from birds were analyzed. Nine nymphs of A. aureolatum (2.1%) and four nymphs of I. auritulus (0.7%) collected from vegetation were positive. Five nymphs of A. aureolatum (45.4%), and five pools of larvae (minimum infection rate 13.5%), 18 nymphs (40.9%) and one female (14.3%) of I. auritulus collected on birds were also positive. The remaining samples were negative. The phylogenetic tree generated with fla sequences shows that seven of the eight different haplotypes of Borrelia detected in I. auritulus conform an independent lineage within the Borrelia burgdorferi sensu lato complex together with sequences of Borrelia sp. detected in I. auritulus from Canada and Uruguay. The fla sequences of Borrelia obtained from A. aureolatum and one sequence of a single specimen of I. auritulus conform a phylogenetic group with Borrelia turcica, Borrelia sp. isolated from a tortoise in Zambia, Borrelia spp. detected in Amblyomma maculatum from USA and Amblyomma longirostre from Brazil. The epidemiological risk that implies the infection with Borrelia genospecies associated with I. auritulus seems to be low because this tick is not aggressive to humans, but it helps to maintain borrelial spirochetes in the enzootic transmission cycles. The pathogenicity to humans of the Borrelia found in A. aureolatum is unknown; however, adults of this tick species are known to bite humans. This is the first report of the presence of Borrelia in A. aureolatum. Further investigations are necessary to know the risk of transmission of borreliosis by hard ticks in the study area.

The presence of Borrelia theileri in Argentina.

The presence of Borrelia theileri in Argentina is confirmed after recording the spirochete from a bovine in northern Argentina. The analysis of sequences of the flagellin gene (fla) and length of Borrelia spp. specimens on thick blood films shows that the local isolate clusters within a well-supported clade with B. theileri isolates from different geographical origins, confirming the presence of B. theileri in Argentina. The mean length of 30 specimens of B. theileri was 12.89 µm (standard deviation 2.88 µm, range 9.35-20.16 µm). The only known vector of Borrelia theileri in northern Argentina is the cattle tick Rhipicephalus microplus, therefore Borrelia infection should be regarded as a potential complication of other cattle tick-borne diseases such as babesiosis, especially on cattle introduced from areas free of R. microplus. The possibility of serologic cross-reaction with B. theileri must not be minimized in studies of other spirochaetes in the R. microplus infested region of Argentina.

Especies de garrapatas duras en un área urbana protegida de la Ciudad Autónoma de Buenos Aires / Hard Tick Species in a Protected Urban Area of Buenos Aires City

INTRODUCCIÓN: Las garrapatas (Acari, Ixodida) son artrópodos vectores de gran diversidad de patógenos virales, bacterianos y protozoarios, muchos de los cuales son zoonóticos y emergentes. En Argentina se encuentran numerosas especies de garrapatas duras (familia Ixodidae) que también parasitan al hombre. Las áreas urbanas protegidas son ecosistemas naturales que están dentro de grandes urbes o limitan con ellas. En la Reserva Ecológica Costanera Sur (RECS) de la Ciudad Autónoma de Buenos Aires (CABA) circulan especies silvestres que no tienen presencia en otras áreas de la ciudad, con potenciales hospedadores y vectores. OBJETIVOS: Determinar las especies de garrapatas presentes en la RECS (CABA). MÉTODOS: Se realizó un estudio cuantitativo descriptivo transversal mediante muestreos para colectar garrapatas de vegetación y de hospedadores como perros y distintas especies de roedores. RESULTADOS: En total se recolectaron 1090 garrapatas de la vegetación (454 de la especie Amblyomma aureolatum, 635 Ixodes auritulus y 1 Amblyomma triste) y 67 de los perros (64 A. aureolatum, 2 Rhipicephalus sanguineus sensu lato y 1 A. triste). No se detectaron garrapatas en 200 roedores revisados. CONCLUSIONES: Las especies de garrapatas detectadas en este estudio tienen importancia en salud pública, tanto porque parasitan a humanos (A. aureolatum, A. triste y R. sanguineus s. l.) como porque participan en el ciclo y transmisión de distintos patógenos zoonóticos en distintas regiones del mundo, incluso Argentina.

INTRODUCTION: Ticks (Acari, Ixodida) are arthropod vectors of great diversity of viral, bacterial and protozoan pathogens, many of which are zoonotic and emerging. In Argentina, there are numerous hard tick species (Ixodidae family) which also parasitize humans. Protected urban areas are natural ecosystems located within or near large urban centers. The Costanera Sur Ecological Reserve (RECS) of Buenos Aires city shows a circulation of wild species that have no presence in other areas of the city. There are potential hosts and vectors. OBJECTIVES: A quantitative cross sectional study was carried to detect the species of ticks present in the RECS of Buenos Aires city. METHODS: A quantitative cross - sectional study was carried out by sampling to collect ticks from vegetation and hosts (rodents and dogs). RESULTS: In total, 1090 ticks were collected from the vegetation (454 of the species Amblyomma aureolatum, 635 Ixodes auritulus and 1 Amblyomma triste) and 67 from dogs (64 A. aureolatum, 2 Rhipicephalus sanguineus sensu lato and 1 A. triste). No ticks were detected in 200 examined rodents. CONCLUSIONS: Ticks species detected in this study are important for public health, because they are parasites of humans (A. aureolatum, A. triste and R. sanguineus s. l.) and participate in the cycle and transmission of different zoonotic pathogens in different regions of the world, including Argentina.

Neorickettsia risticii, Rickettsia sp. and Bartonella sp. in Tadarida brasiliensis bats from Buenos Aires, Argentina.

Bats are potential reservoirs of many vector-borne bacterial pathogens. The aim of the present study was to detect species of Anaplasma, Ehrlichia, Neorickettsia, Rickettsia, Borrelia and Bartonella in Brazilian free-tailed bats (Tadarida brasiliensis, Molossidae) from Buenos Aires city, Argentina. Between 2012 and 2013, 61 T. brasiliensis from urban areas of Buenos Aires city were studied. The samples were molecularly screened by PCR and sequencing. Five bats (8.2%) were positive to Neorickettsia risticii, one (1.6%) was positive to Rickettsia sp. and three bats (4.9%) to Bartonella sp. For molecular characterization, the positive samples were subjected to amplification and sequencing of a fragment of p51 gene for N. risticii, a fragment of citrate synthase gene (gltA) for Rickettsia genus and a fragment of gltA for Bartonella genus. Phylogenetic tree was constructed using the maximum-likelihood method. Phylogenetic analysis of N. risticii detect in our study revealed that it relates to findings in the USA West Coast; Rickettsia sp. detected is phylogenetically within R. bellii group, which also includes many other Rickettsia endosymbionts of insects; and Bartonella sp. found is related to various Bartonella spp. described in Vespertilionidae bats, which are phylogenetically related to Molossidae. Our results are in accordance to previous findings, which demonstrate that insectivorous bats could be infected with vector-borne bacteria representing a potential risk to public health. Future research is necessary to clarify the circulation of these pathogens in bats from Buenos Aires.

Molecular characterization of Ehrlichia canis infecting dogs, Buenos Aires.

Canine monocytic ehrlichiosis (CME) is a worldwide potentially fatal tick-borne rickettsial disease of dogs caused by Ehrlichia canis and transmitted by Rhipicephalus sanguineus sensu lato. CME diagnosis includes indirect (serology) and direct (e.g. blood smears and PCR) methods. PCR is more sensitive and specific than direct microscopic examination and positive PCR results confirm infection, whereas positive serologic test results only confirm exposure. The aim of the present study was to perform a molecular characterization of E. canis from canine samples of the Metropolitan Area of Buenos Aires. We studied 223 blood samples of dogs submitted to our institute for CME diagnoses. The samples were initially screened for Anaplasmataceae family by PCR, resulting in 30 positive dogs (13.4%). Subsequently, positive DNAs were analyzed by nested PCR 16S rRNA specific for E. canis or Anaplasma platys, resulting in 15 (6.7%) and 16 (7.2%) positive dogs, respectively. For molecular characterization, samples positive for E. canis were subjected to amplification of a fragment of the dsb and p28 genes. The nucleotide sequences obtained for the dsb fragment resulted in 100% identity with others E. canis found in dogs from different regions of worldwide. The nucleotide sequences obtained for p28 gene resulted in 100% of identity with each other and closely with E. canis str. Jaboticabal (Brazil). Identity with others sequences of E. canis ranged from 76.9 to 79.7%. The occurrence of canine cases molecularly confirmed in Metropolitan Area of Buenos Aires highlights the need for more studies in order to understand epidemiological factors associated with CME, especially the disease transmission dynamic in South America given the existence of two lineages of R. sanguineus sensu lato with different vectorial capacity for transmission of E. canis.

Tricuspid Annular Plane Systolic Excursion in Acute Decompensated Heart Failure: Relevance for Risk Stratification.

BACKGROUND: Although the prognostic value of right ventricular dysfunction in chronic heart failure (HF) has been studied extensively, it remains insufficiently characterized in the setting of acute decompensated HF (ADHF). We sought to assess whether measurement of tricuspid annular plane systolic excursion (TAPSE) or TAPSE-to-estimated pulmonary arterial systolic pressure (ePASP) ratio allows improvement of risk prediction in ADHF. METHODS: Four hundred ninety-nine patients with ADHF were studied. Cox regression analyses were used to analyze the association of TAPSE and TAPSE-to-ePASP ratio with 1-year mortality and logistic regression analyses to analyze the association of the 2 variables of interest with adverse in-hospital outcome (AiHO) (in-hospital death plus worsening HF). RESULTS: During the 365-day follow-up, 143 patients (28.7%) died. At univariable analysis, both TAPSE (P = 0.026) and TAPSE-to-ePASP ratio (P < 0.0001) were significantly associated with 1-year mortality. At multivariable Cox analysis, age (P = 0.0270), ischemic heart disease (P = 0.020), systolic blood pressure (P = 0.006), log N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels (P < 0.0001), serum sodium levels (P = 0.001), and hemoglobin levels (P = 0.001) at admission were independently associated with 1-year mortality. Adjusting for these covariates, neither TAPSE (P = 0.314) nor TAPSE-to-ePASP ratio (P = 0.237) remained independently associated with 1-year mortality. Eighty-three patients (16.6%) had an AiHO. At multivariable logistic regression analysis, the TAPSE-to-ePASP ratio was independently associated with an AiHO (P = 0.024). The association of TAPSE alone or ePASP alone was not statistically significant. CONCLUSIONS: Our data strongly suggest that early assessment of TAPSE or TAPSE-to-ePASP ratio does not improve prediction of 1-year mortality over other key risk markers in ADHF. Nonetheless, the TAPSE-to-ePASP ratio did appear to be independently associated with AiHO.

Serological evidence of Rickettsia and Coxiella burnetii in humans of Buenos Aires, Argentina.

In Buenos Aires city (Argentina), the circulation of these agents has been detected mainly in vectors and animals, few human cases having been described. The aim of our study was to determine the seroprevalence of Rickettsia (spotted fever--SFG--and typhus--TG--groups) and Coxiella burnetii (Q fever agent) in residents of Buenos Aires city. The study involved 99 participants. Rickettsia IgG antibodies against SFG and TG were detected by IFA in 28.3% and 16.2% of serum samples, respectively. SFG titers were mostly 1/64 (53.6%) with a maximum of 1/512 (3.5%) whereas TG titers ranged between 1/64 (62.5%) and 1/256 (6.3%). Only one sample showed a titer of 1/32 for C. burnetii (phases I and II). The circulation of these pathogens in urban areas such as the city of Buenos Aires should be considered by health services, especially at the primary care level.

C-MYC modulation induces responsiveness to paclitaxel in adrenocortical cancer cell lines.

C-MYC is overexpressed in many types of cancer linked to poor prognosis. We examined the c-Myc protein expression in adrenocortical cancer (ACC) cells to investigate the role of this protein in the neoplasm, its involvement in chemotherapy and finally to determine whether c-Myc could be considered a prognostic factor in patients with ACC. H295R and SW13 cell lines were treated with paclitaxel. c-Myc overexpressing cell clones were achieved by transfecting the H295R cell line with the pcDNA3-hMYC plasmid expressing the full-lengh C-MYC coding sequence. The SW13 cell line was transfected with siRNA oligonucleotides for C-MYC. Cell cycle analysis was evaluated by flow cytometry. c-Myc, cyclin B1 and pro caspase expression levels were evaluated by western blot analysis. We found that expression of c-Myc was highly expressed in the SW13 cells, whereas the protein was undetectable in the H295R cells. Different doses of paclitaxel were required in the two ACC cell line to induce a block in the G2 phase, characterized by increased cyclin B1 levels and to induce apoptosis by pro-caspase-3 activation. Interestingly, the silencing of C-MYC mRNA prevented paclitaxel induced apoptosis in SW13 cells, whereas in the H295R cells the overexpression of C-MYC rendered the cells more prone to growth inhibition after paclitaxel exposure. The present study directly demonstrates that C-MYC plays a central role in controlling proliferation in ACC cells after paclitaxel treatment and that c-Myc could be considered as a marker for predicting response to chemotherapeutic agents in ACC cell lines.

Lung function efficacy and symptomatic benefit of olodaterol once daily delivered via Respimat® versus placebo and formoterol twice daily in patients with GOLD 2-4 COPD: results from two replicate 48-week studies.

Two replicate, multicenter, randomized, double-blind, placebo-controlled, parallel-group, Phase III studies investigated the long-term efficacy and safety of once-daily olodaterol via Respimat® versus placebo and formoterol over 48 weeks in patients with moderate to very severe chronic obstructive pulmonary disease receiving usual-care background therapy. Patients received once-daily olodaterol 5 or 10 µg, twice-daily formoterol 12 µg, or placebo. Co-primary end points were forced expiratory volume in 1 second (FEV1) area under the curve from 0-3 hours response, FEV1 trough response, and Mahler transition dyspnea index total score after 24 weeks; secondary end points included St George's Respiratory Questionnaire. Overall, 904 (Study 1222.13) and 934 (Study 1222.14) patients received treatment. Olodaterol significantly improved FEV1 area under the curve from 0-3 hours versus placebo in both studies (with olodaterol 5 µg, 0.151 L and 0.129 L; with olodaterol 10 µg, 0.165 L and 0.154 L; for all comparisons P<0.0001) and FEV1 trough responses versus placebo (0.053-0.085 L; P<0.01), as did formoterol. Primary analysis revealed no significant difference in transition dyspnea index focal score for any active treatment versus placebo. Post hoc analysis using pattern mixture modeling (accounting for discontinuations) demonstrated statistical significance for olodaterol versus placebo. St George's Respiratory Questionnaire total score was significantly improved with olodaterol, but not formoterol, versus placebo. No safety signals were identified from adverse-event or other safety data. Once-daily olodaterol 5 µg and 10 µg is efficacious in patients with moderate to very severe chronic obstructive pulmonary disease on usual-care maintenance therapy, with a satisfactory safety profile.

Molecular characterization of Rickettsia massiliae and Anaplasma platys infecting Rhipicephalus sanguineus ticks and domestic dogs, Buenos Aires (Argentina).

Rickettsioses, ehrlichioses and anaplasmoses are emerging diseases that are mainly transmitted by arthropods and that affect humans and animals. The aim of the present study was to use molecular techniques to detect and characterize those pathogens in dogs and ticks from Buenos Aires city. We studied 207 Rhipicephalus sanguineus ticks and 52 canine blood samples from poor neighborhoods of Buenos Aires city. The samples were molecularly screened for the genera Rickettsia, Ehrlichia, and Anaplasma by PCR and sequencing. DNA of Rickettsia massiliae (3.4%) and Anaplasma platys (13.5%) was detected in ticks and blood samples, respectively. For characterization, the positive samples were subjected to amplification of a fragment of the 190-kDa outer membrane protein gene (spotted fever group rickettsiae) and a fragment of the groESL gene (specific for A. platys). A phylogenetic tree was constructed using the neighbor-joining method, revealing that the sequences were closely related to those of strains from other geographic regions. The results indicate that human and animal pathogens are abundant in dogs and their ticks in Buenos Aires city and portray the potentially high risk of human exposure to infection with these agents, especially in poor neighborhoods, where there is close contact with animals in an environment of poor health conditions.

Hyper-activation of Notch3 amplifies the proliferative potential of rhabdomyosarcoma cells.

Rhabdomyosarcoma (RMS) is a pediatric myogenic-derived soft tissue sarcoma that includes two major histopathological subtypes: embryonal and alveolar. The majority of alveolar RMS expresses PAX3-FOXO1 fusion oncoprotein, associated with the worst prognosis. RMS cells show myogenic markers expression but are unable to terminally differentiate. The Notch signaling pathway is a master player during myogenesis, with Notch1 activation sustaining myoblast expansion and Notch3 activation inhibiting myoblast fusion and differentiation. Accordingly, Notch1 signaling is up-regulated and activated in embryonal RMS samples and supports the proliferation of tumor cells. However, it is unable to control their differentiation properties. We previously reported that Notch3 is activated in RMS cell lines, of both alveolar and embryonal subtype, and acts by inhibiting differentiation. Moreover, Notch3 depletion reduces PAX3-FOXO1 alveolar RMS tumor growth in vivo. However, whether Notch3 activation also sustains the proliferation of RMS cells remained unclear. To address this question, we forced the expression of the activated form of Notch3, Notch3IC, in the RH30 and RH41 PAX3-FOXO1-positive alveolar and in the RD embryonal RMS cell lines and studied the proliferation of these cells. We show that, in all three cell lines tested, Notch3IC over-expression stimulates in vitro cell proliferation and prevents the effects of pharmacological Notch inhibition. Furthermore, Notch3IC further increases RH30 cell growth in vivo. Interestingly, knockdown of Notch canonical ligands JAG1 or DLL1 in RMS cell lines decreases Notch3 activity and reduces cell proliferation. Finally, the expression of Notch3IC and its target gene HES1 correlates with that of the proliferative marker Ki67 in a small cohort of primary PAX-FOXO1 alveolar RMS samples. These results strongly suggest that high levels of Notch3 activation increase the proliferative potential of RMS cells.

Pharmacological inhibition of EZH2 as a promising differentiation therapy in embryonal RMS.

BACKGROUND: Embryonal Rhabdomyosarcoma (RMS) is a pediatric soft-tissue sarcoma derived from myogenic precursors that is characterized by a good prognosis in patients with localized disease. Conversely, metastatic tumors often relapse, leading to a dismal outcome. The histone methyltransferase EZH2 epigenetically suppresses skeletal muscle differentiation by repressing the transcription of myogenic genes. Moreover, de-regulated EZH2 expression has been extensively implied in human cancers. We have previously shown that EZH2 is aberrantly over-expressed in RMS primary tumors and cell lines. Moreover, it has been recently reported that EZH2 silencing in RD cells, a recurrence-derived embryonal RMS cell line, favors myofiber-like structures formation in a pro-differentiation context. Here we evaluate whether similar effects can be obtained also in the presence of growth factor-supplemented medium (GM), that mimics a pro-proliferative microenvironment, and by pharmacological targeting of EZH2 in RD cells and in RD tumor xenografts. METHODS: Embryonal RMS RD cells were cultured in GM and silenced for EZH2 or treated with either the S-adenosylhomocysteine hydrolase inhibitor 3-deazaneplanocin A (DZNep) that induces EZH2 degradation, or with a new class of catalytic EZH2 inhibitors, MC1948 and MC1945, which block the catalytic activity of EZH2. RD cell proliferation and myogenic differentiation were evaluated both in vitro and in vivo. RESULTS: Here we show that EZH2 protein was abnormally expressed in 19 out of 19 (100%) embryonal RMS primary tumors and cell lines compared to their normal counterparts. Genetic down-regulation of EZH2 by silencing in GM condition reduced RD cell proliferation up-regulating p21Cip1. It also resulted in myogenic-like differentiation testified by the up-regulation of myogenic markers Myogenin, MCK and MHC. These effects were reverted by enforced over-expression of a murine Ezh2, highlighting an EZH2-specific effect. Pharmacological inhibition of EZH2 using either DZNep or MC inhibitors phenocopied the genetic knockdown of EZH2 preventing cell proliferation and restoring myogenic differentiation both in vitro and in vivo. CONCLUSIONS: These results provide evidence that EZH2 function can be counteracted by pharmacological inhibition in embryonal RMS blocking proliferation even in a pro-proliferative context. They also suggest that this approach could be exploited as a differentiation therapy in adjuvant therapeutic intervention for embryonal RMS.

Efficacy and safety of the oral p38 inhibitor PH-797804 in chronic obstructive pulmonary disease: a randomised clinical trial.

RATIONALE: Chronic obstructive pulmonary disease (COPD) is a common lung disease leading to progressive decline in lung function. Inhibition of release of inflammatory mediators by p38 inhibitors may be a useful treatment for chronic inflammation of the airways thought to underlie the pathogenesis of the disease. OBJECTIVES: To evaluate the efficacy and safety of PH-797804, a potent and selective p38 inhibitor, in adults with moderate to severe COPD (Global Initiative for Chronic Obstructive Lung Disease stage II/III). METHODS: This was a randomised, adaptive design, double-blind, placebo-controlled, parallel-group, multicentre trial. Patients were initially randomised to placebo, 0.5, 3, 6 or 10 mg PH-797804 once daily and treated for 6 weeks following a 2-week run-in. MEASUREMENTS AND MAIN RESULTS: The primary endpoint was change from baseline in trough forced expiratory volume in 1 s (FEV1) compared with placebo after 6 weeks of treatment. Secondary endpoints included other spirometric parameters, transition dyspnoea index, rescue mediation use, high sensitivity C-reactive protein and symptoms. A total of 230 patients were assigned to treatment; placebo (n=45), 0.5 mg (n=20), 3 mg (n=47), 6 mg (n=70) and 10 mg (n=48). PH-797804 showed a statistically significant improvement in trough FEV1 at week 6 compared with placebo of 0.086 litre (95% Bayesian CI 0.008 to 0.164) and 0.093 litre (95% CI 0.018 to 0·166) at 3 and 6 mg PH-797804, respectively. PH-797804 3 mg and 6 mg showed an improvement in the baseline dyspnoea index/transition dyspnoea index total focal score at week 6. PH-797804 was well tolerated at all doses studied. CONCLUSIONS: PH-797804 demonstrated improvements over placebo in lung function parameters and dyspnoea in patients with moderate to severe COPD. TRIALREGNO: NCT00559910.