Determination of the welfare status of freeroaming dogs in two urban centres in Chile. / Determinación del estado de bienestar en perros callejeros de dos centros urbanos de Chile.

Free-roaming dogs are not only a public health and ethical problem, they are also an environmental and economic one. Although the general belief is that freeroaming dogs are not in good condition, there have been insufficient studies in Chile to address and analyse the issue. The objective of this research was to assess the welfare of free-roaming dogs in the centre of the cities of Santiago and Valparaíso. The evaluation was carried out using an observational method and assessed a total of 554 dogs. The following variables were analysed: body condition, motor impairment, skin condition, respiratory disease, reaction to humans and other variables. In addition, spatial distribution was considered, and age, sex and social behaviour were estimated. When the results were analysed in terms of frequency, it was found that, in Valparaíso (n = 204), 37% of the dogs assessed had compromised welfare (poor or fair), while 63% had positive welfare (good or optimal). In contrast, in Santiago (n = 350), only 21.7% had compromised welfare, while 78.3% had positive welfare. With respect to social behaviour, 55% of the dogs assessed in Valparaíso and 68% of those assessed in Santiago led a solitary lifestyle. Although most of the individuals were in good condition, a high percentage were unable to meet the requirements for them to live in harmony with their environment.

Les chiens de rue ou errants posent des problèmes non seulement de santé publique et d'éthique, mais aussi environnementaux et économiques. La condition générale de ces chiens est intuitivement perçue comme n'étant pas bonne mais au Chili le sujet n'a guère fait l'objet d'analyses approfondies. Les auteurs présentent les résultats d'une étude visant à évaluer le bien-être animal des chiens errants du centre-ville de Santiago et de Valparaíso. Cette évaluation a reposé sur l'observation de 554 chiens au total et sur l'analyse d'un certain nombre de variables dont la condition physique, les difficultés motrices, l'état de la peau et du pelage, la présence de maladies respiratoires et les réactions face à l'être humain. La distribution spatiale des chiens examinés a été consignée, ainsi que l'âge estimé, le sexe et le comportement social de chaque animal. Les résultats ont été analysés en termes de fréquence. À Valparaíso, les observations ont fait état d'un niveau de bien-être compromis (allant de mauvais à médiocre) chez 37 % des chiens (n = 204) et d'un pourcentage de 63 % de chiens présentant un niveau de bien-être satisfaisant (bon à optimal). En revanche, à Santiago (n = 350), 21 % seulement des chiens observés présentaient un état de bien-être compromis tandis que 78,3 % présentaient un état de bien-être satisfaisant. En ce qui concerne le comportement social, 55 % des chiens observés à Valparaíso et 68 % de ceux observés à Santiago avaient un mode de vie solitaire. Si la majorité des chiens se trouvaient en bon état physique, un pourcentage élevé d'entre eux ne parvenaient pas à satisfaire les besoins leur permettant de vivre en équilibre avec leur environnement.

Los perros callejeros constituyen no solo un problema de salud pública y ético, sino también, medioambiental y económico. Si bien se cree que estos perros no se encuentran en buenas condiciones, en Chile los estudios que abordan y analizan este tema son insuficientes. El objetivo de esta investigación fue evaluar el bienestar en perros callejeros en el centro de las ciudades de Santiago y Valparaíso. La evaluación se realizó a través de un método observacional en un total de 554 perros, y se analizaron las siguientes variables: condición corporal, dificultad motora, estado de la piel, enfermedad respiratoria, reacción hacia el humano y otras variables. Además, se consideró la distribución espacial y se estimó la edad, el sexo y la conducta social. Los resultados fueron analizados en términos de frecuencia, y se observó que en Valparaíso (n = 204), el 37% de los perros presentaba un bienestar comprometido (malo - escaso) mientras que el 63% presentaba un bienestar favorable (bueno u óptimo). En cambio, en Santiago (n = 350), sólo un 21,7% presentaba un estado de bienestar comprometido mientras que el 78,3% presentaba un estado de bienestar favorable. Asimismo, respecto a la conducta social, un 55% de los perros evaluados en Valparaíso y un 68% de los evaluados en Santiago mostraron un estilo de vida solitario. Si bien los individuos en su mayoría se encontraban en buenas condiciones, un alto porcentaje no lograba satisfacer las necesidades para estar en equilibrio con su entorno.

[Ejaculation capacity in spinal cord injured patients carrying an endo-urethral stent for incontinentation: Descriptive study]. / Capacité éjaculatoire chez les sujets blessés médullaires porteurs d'une prothèse endo-urétrale d'incontinentation : étude descriptive.

OBJECTIVES: To assess the impact of endo-urethral stent (EUP) of incontinentation in spinal cord injured (SCI) men on ejaculation capacity. FIRST CRITERIA: Presence of ejaculation after EUP. SECONDARY CRITERIA: Modification on ejaculation type, stimulation mode, possibility of sperm conservation in the same population. POPULATION AND METHODS: A descriptive monocentric study including SCI subjects which underwent placement or change of temporary (Mémokath™ or Allium™) or definitive (Mémotherm™) EUP between January 2004 and January 2014 with at least one ejaculatory test. Are identified for each patient: the presence of ejaculation, type of ejaculation, stimulation mode used, number of frozen specimen and results from semen analysis. RESULTS: Over 11 men with tetraplegia, complete for motricity, there were 8 (72%) for whom ejaculation was possible after laying EUP. Of these, 6 (55%) had an antegrade or antero-retrograde ejaculation. It was not possible to make a freezing straw in four of them due to infection of semen (2) or terato-oligo-astenospermia (1) or absence of reproductive project (1). CONCLUSION: In this small sample, it was found that over 70% ejaculation have been preserved after laying EUP and more than 50% with antegrade or antero-retrograde ejaculation. A larger study would identify how EUP may alter the ejaculatory capacity or quality of the ejaculate. LEVEL OF EVIDENCE: 4.

Novel Acanthamoeba 18S rRNA gene sequence type from an environmental isolate.

The free-living amoebae, Acanthamoeba, can act as opportunistic parasites on a wide range of vertebrates and are becoming a serious threat to human health due to the resistance of their cysts to harsh environmental conditions, disinfectants, some water treatment practices, and their ubiquitous distribution. Subgenus classification based on morphology is being replaced by a classification based on the sequences of the 18S rRNA gene with a total of 18 different genotypes (T1-T18). A new environmental strain of Acanthamoeba isolated from a waste water treatment plant is presented in this study as a candidate for the description of the novel genotype T19 after phylogenetic analysis.

Cardiac safety of dual anti-HER2 blockade with pertuzumab plus trastuzumab in early HER2-positive breast cancer in the APHINITY trial.

BACKGROUND: Trastuzumab increases the incidence of cardiac events (CEs) in patients with breast cancer (BC). Dual blockade with pertuzumab (P) and trastuzumab (T) improves BC outcomes and is the standard of care for high-risk human epidermal growth factor receptor 2 (HER2)-positive early BC patients. We analyzed the cardiac safety of P and T in the phase III APHINITY trial. PATIENTS AND METHODS: Left ventricular ejection fraction (LVEF) ≥ 55% was required at study entry. LVEF assessment was carried out every 3 months during treatment, every 6 months up to month 36, and yearly up to 10 years. Primary CE was defined as heart failure class III/IV and a significant decrease in LVEF (defined as ≥10% from baseline and to <50%), or cardiac death. Secondary CE was defined as a confirmed significant decrease in LVEF, or CEs confirmed by the cardiac advisory board. RESULTS: The safety analysis population consisted of 4769 patients. With 74 months of median follow-up, CEs were observed in 159 patients (3.3%): 83 (3.5%) in P + T and 76 (3.2%) in T arms, respectively. Most CEs occurred during anti-HER2 therapy (123; 77.4%) and were asymptomatic or mildly symptomatic decreases in LVEF (133; 83.6%). There were two cardiac deaths in each arm (0.1%). Cardiac risk factors indicated were age > 65 years, body mass index ≥ 25 kg/m2, baseline LVEF between 55% and <60%, and use of an anthracycline-containing chemotherapy regimen. Acute recovery from a CE based on subsequent LVEF values was observed in 127/155 patients (81.9%). CONCLUSIONS: Dual blockade with P + T does not increase the risk of CEs compared with T alone. The use of anthracycline-based chemotherapy increases the risk of a CE; hence, non-anthracycline chemotherapy may be considered, particularly in patients with cardiovascular risk factors.

Molecular characterization of Acanthamoeba isolated in water treatment plants and comparison with clinical isolates.

A total of 116 samples (44 clinical specimens and 72 environmental samples) have been analyzed for the presence of Acanthamoeba. The environmental samples (ESs) were collected from four drinking water treatment plants (DWTP, n=32), seven wastewater treatment plants (n=28), and six locations of influence (n=12) on four river basins from the central area of Spain (winter-spring 2008). Water samples were concentrated by using the IDEXX Filta-Max(®) system. Acanthamoeba was identified in 65 of the 72 ESs by culture isolation (90.3%) and 63 by real-time PCR (87.5%), resulting in all sampling points (100%) positive for Acanthamoeba when considering both techniques and all the time period analyzed. Nine of the 44 clinical specimens were positive for Acanthamoeba. Seventeen Acanthamoeba strains (eight from four DWTP and nine from clinical samples) were also established in axenic-PYG medium. Twenty-four of the ESs and the 17 Acanthamoeba sp. strains were genotyped as T4/1, T4/8, and T4/9. The eight strains isolated from the DWTP samples were inoculated in nude mouse to ascertain their potential pathogenicity in this model. Animals that were inoculated died or showed central nervous system symptoms 9 days post-inoculation. Examination of immunofluorescence-stained brain and lung tissue sections showed multiple organisms invading both tissues, and re-isolation of throphozoites was successful in these tissues of all infected animals. For the first time, potentially pathogenic Acanthamoeba T4 has been detected in 100% of different types of water samples including tap water and sewage effluents in the central area of Spain suggesting a potential health threat for humans especially for the contact lens wearers.

Late recurrence of cervical cancer: a report of 16 cases.

PURPOSE: To analyze the clinical characteristics and outcome of cervical cancer patients presenting late recurrence. MATERIALS AND METHODS: The medical records of 16 patients who were treated between 1974 and 1999 at the Institution and whose cancer recurred after a five-year disease-free interval were reviewed. RESULTS: Mean time from initial therapy to recurrence was 162.5 months (60-360 mean). Smear abnormalities, atypical genital bleeding, abdominal and lumbar pain, and respiratory findings were the most common symptoms and signs associated with late recurrence. Fourteen patients were diagnosed by physical examination. Three of the six patients with local recurrence who were re-irradiated developed a vesico-vaginal fistula. At a median follow-up time of 12.5 months (4-38 mean), 12 patients were alive and the median survival time was 30 months. CONCLUSIONS: Cervical cancer patients surviving free of disease after the fifth year post-treatment are still at risk for relapse and in most of them, the recurrence is suspected by clinical examination alone.

First cases of microsporidiosis in transplant recipients in Spain and review of the literature.

Microsporidia are currently considered emerging pathogens responsible for life-threatening infections in organ transplant recipients. Here, we describe the first cases of intestinal microsporidiosis by Enterocytozoon bieneusi genotype D in two non-HIV-infected renal transplant recipients from Spain. Previously reported cases of microsporidiosis in organ transplant recipients have also been reviewed, highlighting the necessity of considering organ transplant recipients a risk group for microsporidiosis. A systematic search for these parasites is recommended in cases of persistent diarrhea and in the differential diagnosis of other syndromes, such as chronic fever of unknown etiology.

High-level resistance of Nosema ceranae, a parasite of the honeybee, to temperature and desiccation.

Resistance of Nosema ceranae to different exposure conditions has been evaluated by using Sytox green and DAPI (4',6-diamidino-2-phenylindole) to test spore viability. High thermotolerance at 60 and 35 degrees C and resistance to desiccation were observed. However, a significant decrease in viability after freezing and a rapid degeneration of spores maintained at 4 degrees C were also detected.

Follow-up of antibody avidity in BALB/c mice infected with Toxocara canis.

In human Toxocara canis infection, an association has been shown between high IgG avidity in the chronic phase and low IgG avidity in recently acquired toxocarosis. The evolution of the antibody response in terms of avidity has been carried out through a T. canis infection in BALB/c mice. Infection with T. canis embryonated eggs (EE) was carried out with single doses (SD) of 6, 12, 50, 100, 200 or 1000 EE/mouse and with multiple doses (MD) of 200 and 1000 EE. Specific antibodies against T. canis (IgM+G, IgG, IgG1 and IgM) were detected by ELISA and Western Blot (WB) techniques in the presence and absence of urea. With the ELISA method, an increase in the avidity index (AI) of around 50% was detected from days 40-80 p.i. to the end of the study, with all the doses studied. The WB method showed the presence of high avidity antibodies bound to 100 kDa and 75 kDa T. canis proteins in all the cases when the IgM+G and the IgG1 antibodies were investigated. Antibodies of variable avidity were observed in those sera that recognized the group of low molecular weight proteins, between 37 kDa and 25 kDa.

Encephalitozoon microsporidia modulates p53-mediated apoptosis in infected cells.

Microsporidia are intracellular obligate parasites which have recently been found to be related to fungi. They have a unique extrusion apparatus that is able to inject the sporoplasm directly into the target cell without using receptors. Encephalitozoon microsporidia are a source of morbidity and mortality in humans. It has been suggested that microsporidia may modulate the host cell cycle and apoptosis. We report here that caspase-3 cleavage is inhibited at different times of Vero cell infection by Encephalitozoon microsporidia and that the phosphorylation and translocation of p53 to the nucleus, previous steps for the activation of this protein, do not occur after infection of Vero cells. Consequently, the transcriptional function of p53 is impaired during the infection cycle as demonstrated by luciferase reporter assays. Thus, to our knowledge, for the first time it is shown that an intracellular parasite may be able to multiply in the host cell without activating the p53 apoptotic pathway of that cell. However, changes in the expression of Bcl-2 or Bax levels were not observed.

Vectorial role of Acanthamoeba in Legionella propagation in water for human use.

Legionella spp. is the causative agent of Legionnaires' disease and is transmitted through aerosols emanating from man-made water systems. Legionella resistance to water treatments has been related to its association with environmental amoebae such as Acanthamoeba. Due to the high presence of this protozoon in Spain and the high rate of notification of Legionnaires' disease of this country, the aims of this work were to study the coexistence of these bacteria and protozoa in water as well as their interaction. The usefulness of Acanthamoeba co-culture for the isolation of environmental Legionella was also studied. For this purpose, 70 water samples were collected in 2011 from three Drinking Water Treatment Plants, three Wastewater Treatment Plants and five Natural Pools in Spain. Acanthamoeba was found by PCR in 87.1% (61/70) samples and, by culture in 85.7% (60/70) samples. Legionella was detected by PCR in 58.6% (41/70) of water samples, in 5.7% (4/70) by agar culture and 75.7% (53/70) by Acanthamoeba co-culture. From the 54 Acanthamoeba water isolates, Legionella was detected in 43 of them independently of Acanthamoeba's genotype (T3, T4 and T11). Legionella feeleii, Legionella birminghamiensis, Legionella gresilensis/berliardensis, Legionella fairfieldensis, Legionella drozanski and Legionella falloni were identified. In conclusion, our results showed that environmental Acanthamoeba is infected by Legionella to a high percentage, and due to its ubiquity, high resistance and its pathogenic potential per se, new methods for its elimination should be studied. Also, the high effectivity of Acanthamoeba co-culture for Legionella detection has been shown.

Idiotypic replica of a Toxocara canis excretory/secretory antigen epitope.

This study describes the production, characterization and use of an anti-idiotype serum raised against the monoclonal antibody TC-1 which recognizes a T. canis excretory/secretory antigen (ES Ag) epitope. Anti-idiotypic (anti-Id or Ab2) antibodies were produced in rabbits using TC-1 F(ab')2 fragments; these anti-Id inhibited ES Ag binding to biotinylated TC-1, and also inhibited a larval microprecipitation assay using TC-1. Assays show that the Ab2 beta or "internal image" of a T. canis ES Ag epitope was obtained. The antibodies have been used as an idiotypic copy of ES Ag in a diagnostic ELISA for murine toxocariosis. Affinity-purified anti-Id antibodies were used to raise a homologous anti-anti-Id (Ab3) response in rabbits. Antibody formation was followed in the sera of BALB/c mice inoculated with embryonated eggs of T. canis during a 12-month infestation. A 3-week latency period was observed before specific anti-TC-1 epitope antibodies were detected. High levels were reached at 7 weeks post-inoculation with a maximum at the ninth month, and were then maintained until the end of the experiment. The results show the possible utility of anti-Id antibodies as an ES Ag molecular replica.

Persistence of immune response in human toxocariasis as measured by ELISA.

The antibody titer was followed in a group of patients, clinically diagnosed with toxocariasis, during a 5 year period. We observed that larvae can survive for at least 5 years in humans. Antigenic stimulation was enough to keep high levels of immunoglobulins over this period. Antibody levels decreased slowly and this pattern is similar to that shown by animal models.

The role of brain peptides in neuroimmunomodulation.

Since neuroimmunomodulation is brought about in part, at least, by secretion of pituitary hormones involved in stress and immune responses, we review briefly the hypothalamic control of the release of ACTH, growth hormone, and prolactin. The release of ACTH is controlled particularly by corticotropin-releasing factor (CRF), but vasopressin has intrinsic releasing activity and potentiates the action of CRF at both hypothalamic and pituitary levels. Oxytocin may even potentiate the action of CRF, but has little, if any, ACTH-releasing activity by itself. In addition, epinephrine may augment responses to the CRFs. In contrast, growth hormone is under dual control by growth-hormone-releasing factor (GRF) and somatostatin, and prolactin is under multifactorial control by a series of inhibitors and stimulators. Dopamine is accepted as a physiological prolactin-inhibiting factor (PIF), but probably GABA and possibly acetylcholine as well are PIFs. There is good evidence for a peptide PIF as well. There are a number of prolactin-releasing factors (PRFs) which include oxytocin, vasoactive intestinal polypeptide, PHI and TRH. Several other peptides can also release prolactin, including angiotensin II. In response to stress there is a complex interaction of peptides intrahypothalamically. CRF augments its own release by an ultra short-loop positive feedback, and there is negative ultra short-loop feedback of GRF and somatostatin. Vasopressin appears to augment CRF release as well as to act directly on the pituitary, and there are complex interactions of various peptides to influence prolactin and GH release.

Intracerebroventricular injection of naloxone blocks melatonin-dependent brain [3H]flunitrazepam binding.

The pineal hormone melatonin modulates the brain benzodiazepine binding sites and its circadian rhythm. In the present study the effects of intracerebroventricular (i.c.v.) administration of naloxone (10-20 ng), alone or in association with melatonin and/or beta-endorphin, on [3H]flunitrazepam ([3H]FNZ) binding to the rat cerebral cortex of hypophysectomized rats was investigated. Melatonin (10-20 ng), beta-endorphin (10-20 ng), and melatonin + beta-endorphin (10-20 ng of each compound) all increased [3H]FNZ binding to a similar extent and in a dose-related manner. The effects of melatonin (10 ng) on [3H]FNZ binding were prevented by simultaneous injection with the specific opioid antagonist naloxone. Naloxone also blocks, although to a lesser extent, the effects of beta-endorphin and of melatonin + beta-endorphin injections. Moreover, naloxone blocks the hypophysectomy-dependent increase in [3H]FNZ binding. These results implicate the modulation of melatonin-dependent changes on brain benzodiazepine receptors by opioid peptides.

Suppressive effect of simultaneous injection of ACTH1-10 and beta-endorphin on brain [3H]flunitrazepam binding.

Seven-day hypophysectomized rats were intracerebroventricularly (i.c.v.) injected with beta-endorphin, ACTH1-10 or beta-endorphin+ACTH1-10 (10-20 ng of each compound) and the [3H]flunitrazepam ([3H])FNZ) binding to the rat cerebral cortex of hypophysectomized rats was assayed one hour later. The i.c.v. injection of ACTH1-10 (10-20 ng) or beta-endorphin (10-20 ng) significantly increased [3H]FNZ binding to a similar extent. The effect of i.c.v. injection of ACTH1-10 on brain binding was blunted by simultaneous beta-endorphin administration at the same doses. The i.c.v. naloxone injection (10-20 ng) did not modify the effect of ACTH1-10 (10 ng) on [3H]FNZ binding, but counteracted, in a dose-related manner, the blocking effect of beta-endorphin on ACTH1-10-dependent brain [3H]FNZ binding. The results suggest the existence of an opioid-melanopeptide integration to control brain benzodiazepine receptors.

Characterization of ouabain high-affinity binding to rat cerebral cortex. Modulation by melatonin.

High-affinity [3H]ouabain binding to membrane preparations of rat cerebral cortex was examined using a rapid filtration procedure. At 37 degrees C, binding reached equilibrium in about 60 min. Scatchard analyses of the data at equilibrium revealed a single population of binding sites with a dissociation constant of KD = 3.1 +/- 0.36 nM and a binding site concentration of Bmax = 246.4 +/- 18.4 fmol/mg protein. Kinetic analyses of the association and dissociation curves indicated a kinetic KD = 4.6 nM, which is in good agreement with the value obtained at equilibrium. When various digitalis compounds were tested for their ability to inhibit [3H]ouabain binding, the following Ki values (nM) were obtained: ouabain (3.9); digoxin (18); acetyl-digitoxin (66); k-strophanthin (95); digitoxin (236). When melatonin was added to the incubation medium, the ability of ouabain to inhibit [3H]ouabain binding increased in a dose-related manner to yield the following Ki values (nM): melatonin 10 nM (2); melatonin 20 nM (1.2); melatonin 40 nM (0.8). These data suggest the existence in the rat cerebral cortex of high-affinity ouabain binding sites which may be a locus for the molecular action of melatonin.

Melatonin counteracts pinealectomy-dependent decreases in rat brain [3H]flunitrazepam binding through an opioid mechanism.

The effect of intracerebroventricular (i.c.v.) injection of melatonin and/or beta-endorphin on the [3H]flunitrazepam binding sites in the cerebral cortex of pinealectomized or superior cervical ganglionectomized rats was studied. Pinealectomy decreased the maximum concentration of benzodiazepine receptors (Bmax) without affecting the dissociation constant (KD), while melatonin, ineffective in control animals, counteracted the effect of pinealectomy. Intracerebroventricular injection of beta-endorphin increases Bmax in both control and pinealectomized animals, the effect being significantly higher in the latter. Simultaneous i.c.v. injection of melatonin + beta-endorphin did not further increase Bmax in any group, whereas i.c.v. injection of naloxone significantly blocked the effects of melatonin and/or beta-endorphin administration. Pineal sympathetic denervation produced a significant increase in Bmax and KD, whereas i.c.v. injection of melatonin further increased the former, restoring KD to control values. Neither i.c.v. administration of beta-endorphin or melatonin + beta-endorphin significantly modified the ganglionectomy-dependent increase in Bmax, although both treatments restored KD to control values. Naloxone administration had no effect on beta-endorphin- and melatonin + beta-endorphin-treated ganglionectomized groups, but counteracted the increased effect of melatonin on Bmax in ganglionectomized animals.

Pinealectomy increases ouabain high-affinity binding sites and dissociation constant in rat cerebral cortex.

The effect of the pineal gland on the ouabain high-affinity binding sites (Kd = 3.1 +/- 0.4 nM, Bmax = 246.4 +/- 18.4 fmol/mg protein) in rat cerebral cortex was studied. Pinealectomy increased Bmax (940.7 +/- 42.8 fmol/mg protein) and Kd (7.6 +/- 1.5 nM) while melatonin injection (100 micrograms/kg b.wt.) counteracted these effects, restoring kinetic parameters (Kd = 1.9 +/- 0.05 nM; Bmax = 262.2 +/- 29.6 fmol/mg prot) to control values. Melatonin activity on ouabain binding in vitro did not depend upon a direct effect on the binding sites themselves. However, in competition experiments, melatonin increased binding affinity of ouabain as shown by the decreased IC50 values.

Cross-reactivity between Anisakis simplex sensitization and visceral larva migrans by Toxocara canis.

The aim of this work was to study cross-reactivity in the diagnosis of two related ascaridosis. Nineteen patients diagnosed with recidivous acute urticaria (RAU) caused by Anisakis simplex and 26 patients diagnosed with visceral larva migrans (VLM) caused by Toxocara canis were studied employing commercial diagnostic kits and "in house" assay kits. Cross-reactivity observed was greater when using "in house" assay kits, suggesting that T. canis excretory-secretory antigens were not only recognized by antibodies from patients with RAU but with greater intensity compared to the A. simplex excretory-secretory antigens.