A minimally invasive procedure for blood extraction from Xenopus laevis allows follow up studies without euthanasia.

Blood extraction is extremely important for the development of scientific research; however, the existing methods for amphibian´s blood sampling are invasive, mainly leading to the euthanasia of the animal. Therefore, less intrusive methods that allow the obtention of multiple samples from the same individual, are needed as an alternative to the common methods available. Hence, the aim of this study was to propose a minimally invasive method for obtaining blood from the hind leg of Xenopus laevis, that allows continuous sampling without compromising the wellbeing of the organisms. With this method, it was possible to extract blood and plasma from adults and juveniles, and the amount of sample was enough to perform biochemical and molecular assays to assess the viability of the blood. The results also revealed that this method is a convenient alternative to obtain blood without affecting the welfare of the experimental organisms, avoiding the cull of the animals, and the samples are viable for their use in follow up studies.

Patient preferences in chronic immune-mediated inflammatory diseases potentially treated with biological drugs: discrete choice analysis using real-world data analysis.

OBJECTIVES: Immune-mediated inflammatory diseases (IMIDs) represent a high burden due to their chronicity, high prevalence, and associated comorbidities. Chronic patients' preferences must be considered in IMIDs treatment and follow-up. The objective of this study was to further understand patient's preferences in private settings. METHODS: A literature review was performed to choose the most relevant criteria for patients. A D-efficient discrete choice experiment was designed to elicit preferences of adult patients with IMIDs and potential biological treatment prescription. Participants were collected from private practices (rheumatology, dermatology, and gastroenterology) from February to May 2022. Patients chose between option pairs, characterized by six health-care attributes, as well as monthly out-of-pocket drug price. Responses were analyzed through a conditional logit model. RESULTS: Eighty-seven patients answered the questionnaire. The most frequent pathologies were Rheumatoid Arthritis (31%) and Psoriatic Arthritis (26%). The most relevant criteria were choosing the preferred physician (OR 2.25 [SD0.26]); reducing time until visit with specialist (OR 1.79 [SD0.20]), access through primary care (OR 1.60 [SD0.08]), and an increase in monthly out-of-pocket price from 100€ to 300€ (OR 0.55 [SD0.06]) and to 600€ (OR 0.08 [SD0.02]). CONCLUSIONS: Chronic IMIDs patients showed a preference toward a faster, personalized service, even with a trade-off in terms of out-of-pocket price.

Simultaneous nitrification/denitrification and desulfurization of wastewater polluted with ammonium, COD and sulfide: effectiveness of a new up-flow vertical hybrid reactor.

Industrial wastewater discharges pose an environmental risk. Here, the effectiveness of an up-flow vertical hybrid system, operating with synthetic and industrial wastewater was investigated, as a new approach to perform nitrification/denitrification and desulfurization within a single reactor. The hybrid reactor is divided in two reaction zones, the oxic and anoxic. The removal of chemical oxygen demand (COD), ammonium, and sulfide was investigated, highlighting changes in microbial diversity. The reactor was evaluated at hydraulic residence time (HRT) of 1.6 days, and its performance throughout 180 days is presented in four stages. In stages I-II, high COD and ammonium removal was obtained with synthetic wastewater. In stage-III, sulfide-rich synthetic wastewater did not alter the system, attaining COD, ammonium, and sulfide removal efficiencies of 81, 99.5, and 99.7%, respectively. In the last stage, a mixture of effluents was fed into the reactor at loading rates of 277 mg COD/L-d, 46.5 mg NH4 +-N /L-d, and 15 mg HS--S /L-d. Sulfide and ammonium removals were 100% and 99.9%, respectively. However, low COD removal was observed, being of 51%, and the system removed 97% in terms of BOD5. The structure and microbial diversity also changed. Sulfide feeding, induced the proliferation of sulfur oxidizers like Thiomiscropira and Thiobacillus. Industrial wastewater enhanced the abundance of Pseudomonas (15.53%) and favored the proliferation of new bacteria of the genus Truepera (2.98%) and Alicyclipilus (7.56%). This is the first study reporting simultaneous nitrification/denitrification and desulfurization to remove ammonium, COD and sulfide from complex industrial wastewater using an up-flow vertical hybrid reactor.

Tuberculosis in a Spanish cohort of children living with HIV: the CHOTIS study (Childhood HIV & TB study).

BACKGROUND: Tuberculosis (TB) is the leading opportunistic infection in children with human immunodeficiency virus (HIV), but is uncommon in low prevalence regions. We aim to describe the changing epidemiology and clinical presentation of TB-HIV co-infection in a cohort of HIV-infected children in Spain.METHODS: Children diagnosed with TB between 1995 and 2016 in the paediatric HIV cohort were identified. The incidence and clinical presentation were compared in three periods: 1995-1999 (P1, before initiation of combined antiretroviral therapy, cART), 2000-2009 (P2, increase in immigration), and 2010-2016 (P3, decrease in immigration).RESULTS: We included 29 TB cases among 1183 children aged <18 years (2.4%, 243/100 000 person-years). The proportion was stable in P1 and P2 (1.3%), but decreased in P3 (0.8%). The median age at TB diagnosis was 6.4 years (IQR 4-10.6); most children in P3 were aged >10 years (20% vs. 23.1% vs. 83.3%, P = 0.01). TB was diagnosed at HIV presentation in 11/29 children (37.9%). Foreign-born children accounted for respectively 0%, 8% and 67% of the total number of children in each period (P ≤ 0.0001). One third had extrapulmonary TB; four children died (13.8%).CONCLUSION: In our cohort, the incidence of TB-HIV co-infection decreased with decline in immigration. In regions with adequate cART coverage and low TB transmission, paediatric TB-HIV coinfection is uncommon, but associated with significant morbidity. Strategies for TB surveillance, diagnosis and treatment in this vulnerable population should be reinforced.

Clinical characteristics, prevalence, and factors related to delirium in children of 5 to 14 years of age admitted to intensive care. / Características clínicas, prevalencia y factores asociados al delirium en niños de 5 a 14 años de edad ingresados en cuidados intensivos.

OBJECTIVE: To evaluate the clinical characteristics, prevalence and factors associated with delirium in critical patients from 5 to 14 years of age. DESIGN: An analytical, cross-sectional observational study was made. Delirium was assessed with the Pediatric-Confusion Assessment Method for the Intensive Care Unit (pCAM-ICU) and motor classification was established with the Delirium Rating Scale Revised-98. SETTING: A pediatric Intensive Care Unit. PATIENTS: All those admitted over a one-year period were assessed during the first 24-72h, or when possible in deeply sedated patients. EXCLUSION CRITERIA: Patients in stupor or coma, with severe communication difficulty, subjected to deep sedation throughout admission, and those with denied consent. RESULTS: Twenty-nine of the 156 assessed patients suffered delirium (18.6%) and 55.2% were hypoactive. The neurocognitive alterations evaluated by the pCAM-ICU were similar in the three motor groups. Intellectual disability (OR=17.54; 95%CI: 3.23-95.19), mechanical ventilation (OR=18.80; 95%CI: 4.29-82.28), liver failure (OR=54.88; 95%CI: 4.27-705.33), neurological disease (OR=4.41; 95%CI: 1.23-15.83), anticholinergic drug use (OR=3.23; 95%CI: 1.02-10.26), different psychotropic agents (OR=4.88; 95%CI: 1.42-16.73) and tachycardia (OR=4.74; 95%CI: 1.21-18.51) were associated to delirium according to the logistic regression analysis. CONCLUSION: The frequency of delirium and hypoactivity was high. It is therefore necessary to routinely evaluate patients with standardized instruments. All patients presented with important neurocognitive alterations. Several factors related with the physiopathology of delirium were associated to the diagnosis; some of them are modifiable through the rationalization of medical care.

Impact of specific modes of circumferential resection margin involvement in rectal cancer local recurrence: A retrospective study.

PURPOSE: To compare the local recurrence rate and overall survival between patients with circumferential resection margin (CRM) involvement by direct tumor spread and by a tumor within a lymph node. METHODS: A total of 776 patients diagnosed with rectal cancer underwent rectal resection. Patients with CRM involvement by direct tumor spread were named group A. CRM involvement by tumor within a lymph node formed group B. Patient data, including sex, age, pT, pN, stage I-III versus IV, neoadjuvant radiotherapy, adjuvant chemotherapy, carcinoembryonic antigen, primary tumor location, lymph node retrieval, and need for abdominoperineal resection, were compared between both groups. RESULTS: In total, 10.5% of the patients had CRM involvement. Of these, in 57 cases (7.3%), it was by direct tumor spread and in 19 cases (2.4%) by tumor within a lymph node. Other types of CRM involvement were found in six patients. With a mean follow-up of 32.9 months, 18 patients from group A (31.5%) and one patient from group B (5.2%) suffered a local recurrence. Local recurrence-free survival was significantly higher in patients from group B (P = 0.049). Patients in stage I-III (P = 0.037) and from group B ( P = 0.049) had better overall survival. CONCLUSION: Patients with CRM involvement by tumor within a lymph node have a low risk of local recurrence and better overall survival than patients with CRM involvement by direct tumor spread.

Chronic stress of rainbow trout Oncorhynchus mykiss at high altitude: a field study.

The stress response of Oncorhynchus mykiss in high-altitude farms in central Mexico was investigated over two seasons: the cool (9·1-13·7° C) dry winter season, and the warmer (14·7-15·9° C), wetter summer season. Fish were subjected to an acute stress test followed by sampling of six physiological variables: blood cortisol, glucose, lactate, total antioxidant capacity, haemoglobin concentration and per cent packed cell volume (VPC %). Multivariate analyses revealed that lactate and total antioxidant capacity were significantly higher in the summer, when water temperatures were warmer and moderate hypoxia (4·9-5·3 mg l(-1) ) prevailed. In contrast, plasma cortisol was significantly higher in the winter (mean ± s.e.: 76·7 ± 4·0 ng ml(-1) ) when temperatures were cooler and dissolved oxygen levels higher (6·05-7·9 mg l(-1) ), than in the summer (22·7 ± 3·8 ng ml(-1) ). Haemoglobin concentrations (mg dl(-1) ) were not significantly different between seasons, but VPC % was significantly higher in the summer (50%) than in the winter (35%). These results suggest that in summer, effects of high altitude on farmed fish are exacerbated by stresses of high temperatures and hypoxia, resulting in higher blood lactate, increased total antioxidant capacity and elevated VPC % levels.

[Spanish Society for Pediatric Infectious Diseases guidelines on tuberculosis in pregnant women and neonates (i): Epidemiology and diagnosis. Congenital tuberculosis]. / Guía de la Sociedad Española de Infectología Pediátrica sobre tuberculosis en la embarazada y el recién nacido (I): epidemiología y diagnóstico. Tuberculosis congénita.

Tuberculosis (TB) screening in pregnancy using tuberculin skin test (TST) is recommended in case of symptoms of TB disease, close contact with a patient with infectious TB, or high risk of developing active disease. The new interferon gamma release assay (IGRA) tests are recommended in BCG-vaccinated pregnant women with positive TST and no known risk factors for TB, and in those immunocompromised, with clinical suspicion of TB but negative TST. TB diagnosis is difficult due to the non-specific symptoms, the increased frequency of extrapulmonary disease, the delay in radiological examinations, and the high rate of tuberculin anergy. Neonatal TB can be acquired in utero (congenital TB), or through airborne transmission after delivery (postnatal TB). Congenital TB is extremely rare and does not cause fetal malformations. It may be evident at birth, although it usually presents after the second week of life. In newborns with no family history of TB, the disease should be considered in cases of miliary pneumonia, hepatosplenomegaly with focal lesions, or lymphocytic meningitis with hypoglycorrhachia, especially in those born to immigrants from high TB-burden countries. TST is usually negative, and IGRAs have lower sensitivity than in older children. However, the yield of acid-fast smear and culture is higher, mostly in congenital TB. Molecular diagnosis techniques enable early diagnosis and detection of drug resistance mutations. There is a substantial risk of disseminated disease and death.

[Spanish Society for Pediatric Infectious Diseases guidelines on tuberculosis in pregnant women and neonates (ii): Prophylaxis and treatment]. / Guía de la Sociedad Española de Infectología Pediátrica sobre tuberculosis en la embarazada y el recién nacido (ii): profilaxis y tratamiento.

In pregnant women who have been exposed to tuberculosis (TB), primary isoniazid prophylaxis is only recommended in cases of immunosuppression, chronic medical conditions or obstetric risk factors, and close and sustained contact with a patient with infectious TB. Isoniazid prophylaxis for latent tuberculosis infection (LTBI) is recommended in women who have close contact with an infectious TB patient or have risk factors for progression to active disease. Otherwise, it should be delayed until at least three weeks after delivery. Treatment of TB disease during pregnancy is the same as for the general adult population. Infants born to mothers with disseminated or extrapulmonary TB in pregnancy, with active TB at delivery, or with postnatal exposure to TB, should undergo a complete diagnostic evaluation. Primary isoniazid prophylaxis for at least 12 weeks is recommended for those with negative diagnostic tests and no evidence of disease. Repeated negative diagnostic tests are mandatory before interrupting prophylaxis. Isoniazid for 9 months is recommended in LTBI. Treatment of neonatal TB disease is similar to that of older children, but should be maintained for at least 9 months. Respiratory isolation is recommended in congenital TB, and in postnatal TB with positive gastric or bronchial aspirate acid-fast smears. Separation of mother and infant is only necessary when the mother has received treatment for less than 2 weeks, is sputum smear-positive, or has drug-resistant TB. Breastfeeding is not contraindicated, and in case of mother-infant separation expressed breast milk feeding is recommended.

Bartonella chomelii is the most frequent species infecting cattle grazing in communal mountain pastures in Spain.

The presence of Bartonella spp. was investigated in domestic ungulates grazing in communal pastures from a mountain area in northern Spain, where 18.3% (17/93) of cattle were found to be positive by PCR combined with a reverse line blot (PCR/RLB), whereas sheep (n = 133) or horses (n = 91) were found not to be infected by this pathogen. Bartonella infection was significantly associated with age, since older animals showed a higher prevalence than heifers and calves. In contrast to other studies, B. chomelii was the most frequent species found in cattle (14/17), while B. bovis was detected in only three animals. Moreover, 18 B. chomelii isolates and one B. bovis isolate were obtained from nine animals. Afterwards, B. chomelii isolates were characterized by a multilocus sequence typing (MLST) method which was adapted in this study. This method presented a high discrimination power, identifying nine different sequence types (STs). This characterization also showed the presence of different STs simultaneously in the same host and that STs had switched over time in one of the animals. In addition, B. chomelii STs seem to group phylogenetically in two different lineages. The only B. bovis isolate was characterized with a previously described MLST method. This isolate corresponded to a new ST which is located in lineage I, where the B. bovis strains infecting Bos taurus subsp. taurus are grouped. Further studies on the dynamics of Bartonella infection in cattle and the potential ectoparasites involved in the transmission of this microorganism should be performed, improving knowledge about the interaction of Bartonella spp. and domestic ungulates.

Visual function assessment in simulated real-life situations in patients with age-related macular degeneration compared to normal subjects.

PURPOSE: To evaluate visual function variations in eyes with age-related macular degeneration (AMD) compared to normal eyes under different light/contrast conditions using a time-dependent visual acuity testing instrument, the Central Vision Analyzer (CVA). METHODS: Overall, 37 AMD eyes and 35 normal eyes were consecutively tested with the CVA after assessing best-corrected visual acuity (BCVA) using ETDRS charts. The CVA established visual thresholds for three mesopic environments (M1 (high contrast), M2 (medium contrast), and M3 (low contrast)) and three backlight-glare environments (G1 (high contrast, equivalent to ETDRS), G2 (medium contrast), and G3 (low contrast)) under timed conditions. Vision drop across environments was calculated, and repeatability of visual scores was determined. RESULTS: BCVA significantly reduced with decreasing contrast in all eyes. M1 scores for BCVA were greater than M2 and M3 (P<0.001); G1 scores were greater than G2 and G3 (P<0.01). BCVA dropped more in AMD eyes than in normal eyes between M1 and M2 (P=0.002) and between M1 and M3 (P=0.003). In AMD eyes, BCVA was better using ETDRS charts compared to G1 (P<0.001). The drop in visual function between ETDRS and G1 was greater in AMD eyes compared to normal eyes (P=0.004). Standard deviations of test-retest ranged from 0.100 to 0.139 logMAR. CONCLUSION: The CVA allowed analysis of the visual complaints that AMD patients experience with different lighting/contrast time-dependent conditions. BCVA changed significantly under different lighting/contrast conditions in all eyes, however, AMD eyes were more affected by contrast reduction than normal eyes. In AMD eyes, timed conditions using the CVA led to worse BCVA compared to non-timed ETDRS charts.

[Emergence of invasive pneumococcal disease caused by non-vaccine serotypes in the era of the 7-valent conjugate vaccine]. / Serotipos no vacunales emergentes causantes de enfermedad neumocócica invasora en la era de la vacuna 7-valente.

INTRODUCTION: There has been an increased incidence in invasive pneumococcal disease (IPD) produced by non-vaccine serotype (NVS) of Streptococcus pneumoniae after the introduction of PCV7. Our objective was to describe the epidemiological, clinical and microbiological characteristics of IPD caused by NVS in a tertiary hospital in Madrid. PATIENTS AND METHODS: Retrospective (1998-2004) and prospective (2005-2009) study evaluating IPD caused by NVS in children. The study was divided into three periods: P1 (1998-2001) when PCV7 was not commercialized; P2 (2002-2005) with 40% vaccine coverage among children; and P3 (2006-2009) when the vaccine was added to the Childhood Immunization Schedule in Madrid. RESULTS: We analyzed 155 cases of IPD. One hundred and fifty of these isolates were serotyped (100 were NVS). There was an increase in the prevalence of IPD from P1 (31%) to P2 (54%) and P3 (91%). The most relevant emerging serotypes were 19A, 7F, 1, 5, 3 and 15C. The most significant clinical syndromes produced by some specific serotypes were as follows: lower respiratory tract infection (LRTI) by serotypes 1, 3, 5 and 15C; LRTI, primary bacteremia and meningitis by serotype 19A; and primary bacteremia by serotype 7F (66%). The large majority (83.8%) of NVS were sensitive to penicillin. CONCLUSIONS: There has been an increased prevalence of IPD caused by NVS since the introduction of PCV7. These changes should prompt the introduction of new pneumococcal vaccines, which include most of the NVS, in the childhood immunization calendar to prevent IPD in children.

Influence of the brain sexual differentiation process on despair and antidepressant-like effect of fluoxetine in the rat forced swim test.

Sex differences exist in the depressive disorder prevalence and response to treatment. Several studies suggest that females respond better than males to the action of selective serotonin reuptake inhibitors (SSRIs), suggesting that gonadal hormones modulate mood and the response to these drugs. Sexual steroid hormones exert organizational actions (perennial and on early development) and activational effects (transient and on differentiated tissues). The aim of this study was to analyze sex differences in the forced swim test (FST) in animals without treatment and after fluoxetine (FLX, 0, 2.5, 5.0 and 10.0mg/kg). Initially, we compared male and female adult rats under control conditions or after altering their sexual differentiation process (at day 5 postnatally, PN, 60µg of testosterone propionate to females and male castration to induce or preclude masculinization, respectively). To further analyze if the sex differences depend on organizational or activational steroid hormone action we tested the same animals before and after adult gonadectomy. To prevent variations depending upon the estrous cycle, control and masculinized females were tested in estrus. Control females showed lower immobility and required lower doses of FLX (5mg/kg), to show an antidepressant-like effect, than males (10mg/kg), even after adult gonadectomy. In control males adult orchidectomy prevented FLX's action. Neonatally masculinized females exhibited analogous levels of immobility than control ones; before ovariectomy they responded to FLX similar to controls, but after the surgery they did not respond to fluoxetine. Neonatally orchidectomized males exhibited similar immobility values and response to FLX than control females. The findings suggest that the sex difference in despair depends on the hormones organizational effects and, in males, the response to FLX relies on organizational and activational actions.

[Increase in the incidence of invasive pneumococcal disease caused by serotype 19A prior to the implementation of the expanded pneumococcal vaccines]. / Aumento de la incidencia de enfermedad neumocócica invasora producida por el serotipo 19A previo a la introducción de las vacunas neumocócicas ampliadas.

OBJECTIVE: To describe the epidemiology, clinical syndromes and microbiological characteristics of serotype 19A as the main cause of invasive pneumococcal disease (IPD) in children admitted to a tertiary hospital in Spain. METHODS: A retrospective (1998-2004) and prospective (2005-2009) study was conducted on children with IPD produced by serotype 19A. The study was divided into three periods (P): P1 (1998-2001) when PCV7 had not been commercialized; P2 (2002-2005) with 40% vaccine coverage among children; and P3 (2006-2009) when the vaccine was added to the Childhood Immunization Schedule in Madrid. RESULTS: A total of 155 isolates of Streptococcus pneumoniae (SP) producing IPD were analysed, with 21 of them being serotype 19A (14%). An increased prevalence of serotype 19A was found: 2/45 cases (4.4%) in P1, 3/41 cases (7.3%) in P2 and 16/69 cases (23.2%) in P3. It occurred mostly in children younger than 2 years (16/21; 76%). This serotype was the main cause of meningitis (5/20; 25%), pleural empyema (3/22; 14%) and bacteraemic mastoiditis (2/4; 50%). Thirteen isolates (61.5%) had an MIC ≥ 0.12µ/ml for penicillin in extra-meningeal infections, and 3 of the 5 isolates causing meningitis (60%) had an MIC ≥ 1µ/ml for cefotaxime. CONCLUSIONS: Serotype 19A was the main causal agent of IPD in the PCV7 era (P3), with high antibiotic resistance rates. This serotype was responsible for all types of IPD, being the main cause of meningitis.

Boldness and aggressiveness in early and late hatched three-spined sticklebacks Gasterosteus aculeatus.

Levels of boldness and the degree of aggressiveness were compared in juvenile three-spined sticklebacks Gasterosteus aculeatus that had hatched early and late in the breeding season. The most striking result found in this study was that early hatched individuals were bolder when exploring a novel environment than were late-hatched individuals. No differences in levels of aggression between early and late hatchlings were found, but a relationship between boldness and aggressiveness was present regardless of hatching date. The implications of these findings are discussed in the light of research on individual variation in behaviour and the development of behavioural syndromes.

[Consensus document by the Spanish Society of Paediatric Infectious Diseases and the advisory committee on vaccines of the Spanish Paediatrics Association on vaccination in immunocompromised children]. / Documento de consenso de la Sociedad Española de Infectología Pediátrica y el Comité Asesor de Vacunas de la Asociación Española de Pediatría para la vacunación en inmunodeprimidos.

Vaccination in immunocompromised infants, children and adolescents is a major aspect in the follow-up of this complex pathology in specific Paediatric Units. Vaccination is also an important prevention tool, as this can, to a certain extent, determine the morbidity and mortality in these patients. This consensus document was jointly prepared by Working Groups of the Spanish Society of Paediatric Infectious Diseases and the Advisory Committee on Vaccines of the Spanish Paediatric Association, who are usually involved in updating the management of vaccinations in immunocompromised children, and reflects their opinions. The consensus specifically summarises indications for vaccination in the following special paediatric populations: Solid organ and haematopoietic transplant-recipients; primary immunodeficiency; asplenic children; non-previously transplanted immunocompromised patients; chronically ill patients; HIV-infected children and also the vaccines recommended for immunodeficient children who travel.

Consistent boldness behaviour in early emerging fry of domesticated Atlantic salmon (Salmo salar): Decoupling of behavioural and physiological traits of the proactive stress coping style.

Individual variation in the way animals cope with stressors has been documented in a number of animal groups. In general, two distinct sets of behavioural and physiological responses to stress have been described: the proactive and the reactive coping styles. Some characteristics of stress coping style seem to be coupled to the time to emerge of fry from spawning redds in natural populations of salmonid fishes. In the present study, behavioural and physiological traits of stress coping styles were compared two and five months after emergence in farmed Atlantic salmon (Salmo salar), using individuals with an early or late time to emerge. Initially, compared to late emerging individuals, early emerging individuals showed a shorter time to resume feeding after transfer to rearing in isolation. Resumption of feeding after isolation was suggested to be related to boldness behaviour, rather than hunger, in the present study. This observation was repeated five months after emergence, demonstrating behavioural consistency over time in this trait. However, in other traits of proactive and reactive stress coping styles, such as social status, resting metabolism or post stress cortisol concentrations, early and late emerging individuals did not differ. Therefore, this study demonstrates that boldness in a novel environment is uncoupled from other traits of the proactive and reactive stress coping styles in farmed salmonids. It is possible that this decoupling is caused by the low competitive environment in which fish were reared. In natural populations of salmonids, however, the higher selection pressure at emergence could select for early emerging individuals with a proactive coping style.