Comparative in silico design and validation of GPS™ CoVID-19 dtec-RT-qPCR test.

AIMS: Providing a ready-to-use reverse transcriptase qPCR (RT-qPCR) method fully validated to detect the SARS-CoV-2 with a higher exclusivity than this shown by early published RT-qPCR designs. METHODS AND RESULTS: The specificity of the GPS™ CoVID-19 dtec-RT-qPCR test by analysis of sequence alignments was approached and compared with other RT-qPCR designs. The GPS™ CoVID-19 dtec-RT-qPCR test was validated following criteria of UNE/EN ISO 17025:2005 and ISO/IEC 15189:2012. Diagnostic validation was achieved by two independent reference laboratories, the Instituto de Salud Carlos III, (Madrid, Spain), the Public Health England (Colindale, London, UK), and received the label CE-IVD. The GPS design showed the highest exclusivity and passed all parameters of validation with strict acceptance criteria. Results from reference laboratories 100% correlated with these obtained by using reference methods and showed 100% of diagnostic sensitivity and specificity. CONCLUSIONS: The CE-IVD GPS™ CoVID-19 dtec-RT-qPCR test, available worldwide with full analytical and diagnostic validation, is the more exclusive for SARS-CoV-2 by far. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering the CoVID-19 pandemic status, the exclusivity of RT-qPCR tests is crucial to avoid false positives due to related coronaviruses. This work provides of a highly specific and validated RT-qPCR method for detection of SARS-CoV-2, which represents a case of efficient transfer of technology successfully used since the pandemic was declared.

Draft Genome Sequence of Aeromonas lusitana sp. nov. Strain DSM 24905T, Isolated from a Hot Spring in Vila-Real, Portugal.

Aeromonas lusitana sp. nov. is an isolate derived from a study aimed at characterizing Aeromonas spp. from water sources used for recreation and agricultural purposes and assessing the implications these organisms have for human and animal health. We present here the 4.52-Mbp draft genome sequence of this novel species.

Draft Genome Sequence of Aeromonas cavernicola sp. nov. DSM 24474T, Isolated from a Cavern Brook in the Moravia Region of the Czech Republic.

Species of the Aeromonas genus can be found in numerous environmental milieus, including various water sources, and some species cause disease in animals. We present here the draft genome sequence for Aeromonas cavernicola DSM 24474T, a novel species isolated from a freshwater brook within a cavern in the Czech Republic.

Phylogenetic analyses of the genus Aeromonas based on housekeeping gene sequencing and its influence on systematics.

The phylogenies derived from housekeeping gene sequence alignments, although mere evolutionary hypotheses, have increased our knowledge about the Aeromonas genetic diversity, providing a robust species delineation framework invaluable for reliable, easy and fast species identification. Previous classifications of Aeromonas, have been fully surpassed by recently developed phylogenetic (natural) classification obtained from the analysis of so-called 'molecular chronometers'. Despite ribosomal RNAs cannot split all known Aeromonas species, the conserved nature of 16S rRNA offers reliable alignments containing mosaics of sequence signatures which may serve as targets of genus-specific oligonucleotides for subsequent identification/detection tests in samples without culturing. On the contrary, some housekeeping genes coding for proteins show a much better chronometric capacity to discriminate highly related strains. Although both, species and loci, do not all evolve at exactly the same rate, published Aeromonas phylogenies were congruent to each other, indicating that, phylogenetic markers are synchronized and a concatenated multigene phylogeny, may be 'the mirror' of the entire genomic relationships. Thanks to MLPA approaches, the discovery of new Aeromonas species and strains of rarely isolated species is today more frequent and, consequently, should be extensively promoted for isolate screening and species identification. Although, accumulated data still should be carefully catalogued to inherit a reliable database.

Synthesis of robust hierarchical silica monoliths by surface-mediated solution/precipitation reactions over different scales: designing capillary microreactors for environmental applications.

A synthetic procedure to prepare novel materials (surface-mediated fillings) based on robust hierarchical monoliths is reported. The methodology includes the deposition of a (micro- or mesoporous) silica thin film on the support followed by growth of a porous monolithic SiO2 structure. It has been demonstrated that this synthesis is viable for supports of different chemical nature with different inner diameters without shrinkage of the silica filling. The formation mechanism of the surface-mediated fillings is based on a solution/precipitation process and the anchoring of the silica filling to the deposited thin film. The interaction between the two SiO2 structures (monolith and thin film) depends on the porosity of the thin film and yields composite materials with different mechanical stability. By this procedure, capillary microreactors have been prepared and have been proved to be highly active and selective in the total and preferential oxidation of carbon monoxide (TOxCO and PrOxCO).

Phylogenetic diversity of Aeromonas from "alheira," a traditional Portuguese meat product.

"Alheira" is a traditional smoked meat sausage produced in the north of Portugal, representing an important economic resource for the region. This meat product has been subjected to research studies with the aim of detecting the presence of common foodborne pathogens, but, to our knowledge, isolation of emerging foodborne Aeromonas from alheira has never been previously described. Present work attempts to evaluate the Aeromonas species diversity of 84 isolates of Aeromonas spp. collected from 32 alheira samples. All presumptive Aeromonas isolates were subjected to genotyping by enterobacterial repetitive intergenic consensus-polymerase chain reaction analysis. The isolates presenting a different pattern were subjected to gyrB gene sequencing for species classification, and the species A. hydrophila, A. salmonicida, A. caviae, A. media, and A. allosaccharophila were identified. The Aeromonas species diversity found has not been previously described in any other meat product evaluated in previous studies. It is also important to highlight the presence of A. hydrophila and A. caviae because they were previously associated with illness in humans, including gastroenteritis.

Phylogenetic identification of Aeromonas from pigs slaughtered for consumption in slaughterhouses at the North of Portugal.

In the present study, 710 isolates of Aeromonas spp. have been collected from pig carcasses, diaphragm muscle, faeces, dehairing equipment and water in slaughterhouses at the North of Portugal. The isolates were obtained from a total of 154 samples. All presumptive Aeromonas isolates were subjected to ERIC-PCR analysis and those which presented a different pattern were taken and the species classified by gyrB gene sequencing. We have found the species A. hydrophila, A. salmonicida, A. bestiarum, A. caviae, A. media, A. veronii, A. allosaccharophila, A. simiae and A. aquariorum. To our knowledge, this extent of Aeromonas species diversity has not been previously described from meat or from the slaughter environment, perhaps due to the unreliability of available identification methods. A noticeable level of isolate redundancy (strains with identical gyrB sequence) from different samples collected in different dates was also obtained, indicating that only a few predominant strains of these species persist at the slaughter system. It is also important to emphasise the presence of Aeromonas species previously associated with illness in man.

Aeromonas rivuli sp. nov., isolated from the upstream region of a karst water rivulet.

Two freshwater isolates (WB4.1-19(T) and WB4.4-101), sharing 99.9 % 16S rRNA gene sequence similarity, were highly related to Aeromonas sobria (99.7 % similarity; 6 bp differences). A phylogenetic tree derived from a multi-locus phylogenetic analysis (MLPA) of the concatenated sequences of five housekeeping genes (gyrB, rpoD, recA, dnaJ and gyrA; 3684 bp) revealed that both strains clustered as an independent phylogenetic line next to members of Aeromonas molluscorum and Aeromonas bivalvium. The DNA-DNA reassociation value between the two new isolates was 89.3 %. Strain WB4.1-19(T) had a DNA-DNA relatedness value of <70 % with the type strains of the other species tested. Phenotypic characterization differentiated the two novel strains from all other type strains of species of the genus Aeromonas. It is concluded that the two new strains represent a novel species of the genus Aeromonas, for which the name Aeromonas rivuli sp. nov. is proposed, with the type strain WB4.1-19(T) (=CECT 7518(T)=DSM 22539(T)=MDC 2511(T)).

[Psychometric properties of the Activities Daily Life Scale (ADL)]. / Caractéristiques psychométriques de l'échelle des activités de la vie quotidienne (AVQ).

OBJECTIVE: Deficits in social functioning are an important core feature of mental health. Recently in France, the Activities Daily Life (ADL) scale has been proposed by the French authorities to assess social functioning for all hospitalized patients in a psychiatric ward. The perspective is to use this scale in the financing and organization of mental health services in France. The ADL scale is a 6-item (dressing/undressing, walking/mobility, eating/drinking, using toilets, behaviour, relationships/communication) heteroquestionnaire completed by a health care professional at the beginning of each hospitalization, assessing functioning of patients suffering from mental health diseases. However, limited consensus exists on this scale. The psychometric properties of the ADL scale have not been assessed. There is a pressing need for detailed examination of its performance. The aim of this study was to explore ADL psychometric properties in a sample of hospitalized patients in a psychiatric ward. METHOD: We retrospectively analyzed data for all episodes of care delivered to hospitalized patients in a psychiatric ward in our French Public Hospital from January 1, 2008 to June 30, 2008. The study involved retrospective review of administrative and medical databases. The following data were collected: age, gender, diagnoses based on the International Classification of Diseases - 10th version, ADL scale and Assessment of Social Self-Sufficiency scale (ASSS). The psychometric properties were examined using construct validity, reliability, external validity, reproducibility and sensitivity to change. Data analysis was performed using SPSS 15.0 and WINSTEP software. RESULTS: A total of 1066 patients completed the ADL scale. Among them, 49.7% were male, mean age was 36.5 ± 10.8, and 83.5% were single. Schizophrenia, schizotypal and delusional disorders (40.0%), mood disorders (27.9%) and mental and behavioural disorders due to psychoactive substance use (12%) were the most common diagnoses. Factor analysis with varimax rotation identified a 2-factor structure accounting for 82% of the total variance. The first dimension (ADL 1) comprised four items and represented personal care activities. The second dimension (ADL 2) comprised two items and represented social functioning. A floor effect was reported for ADL 1 and its unidimensionality was not satisfactory: two items showed an INFIT statistic outside the acceptable range. Internal consistency was satisfactory for the two dimensions: each item achieved the 0.40 standard for item-internal consistency. The correlation of each item with its contributory dimension was higher than with the other (item discriminant validity). Cronbach's alpha coefficients ranged over 0.70 in the whole sample. Concerning external validity, positive correlations were not systematically found between ADL and ASSS dimensions. The score of ADL 1 had medium to high correlations with four dimensions scores of the ASSS, while the score of ADL 2 were not at all or weakly correlated with ASSS dimension scores. Globally, ADL did not cover sentimental life and social relationships. There were statistical associations between ADL and age or gender: women and subjects older than 60 had a higher level of dependency. We didn't find any association with marital status or diagnoses. The ADL scale presented a good reproducibility but was not sensitive to change. CONCLUSION: The psychometric properties of the ADL scale were not sufficient for several parameters such as validity or sensitivity to change, contrary to other available French scales. The use of a heteroquestionnaire rather than a self-administered questionnaire should be discussed by professionals and the French authorities. These results should be taken into account in the use of the ADL scale for the economic and administrative management of psychiatry. Further research should be conducted to confirm these results.

Use of nonabsorbable polymer ligaclip in hand-assisted laparoscopic nephrectomy for living donor.

BACKGROUND: Laparoscopic living donor nephrectomy for kidney transplantation is a technique that began in South America only recently. This procedure offers several advantages compared with open nephrectomy due to the minor pain, better cosmetic results, and shorter length of hospital stay. Herein we have described our experience with the use of nonabsorbable polymer ligaclip (NPL) to control the renal artery, vein, and ureter in hand-assisted laparoscopic donor nephrectomy (HALN). METHODS: We performed a review of 85 HALNs for renal transplantation at our institution between January 2004 and August 2007. We evaluated the preoperative characteristics of the donor, surgical parameters, and complications. RESULTS: Fifty six percent of donors were men. Mean donor age was 34.4 years (range = 18 to 60). Left-sided nephrectomy was performed in 85%. Mean surgical time was 132 minute (range = 90 to 240) and among the last 35 nephrectomies, 120 minute (range = 90 to 180). Mean warm ischemia time was 240 seconds (range = 120 to 420). Conversion rate was 1.1%. Mortality was one case (1.1%) secondary to an episode of massive pulmonary thromboembolism. Mean length of hospital stay was 2.5 days (range 2 to 5) and mean blood loss, 125 mL. No complication related to the NPL was observed. CONCLUSIONS: Laparoscopy living donor nephrectomy was a safe procedure for kidney transplantation. The training and experience of the surgeon was reflected in shorter times of surgery. The NPL was safe and cost-effective, not increasing morbidity of the procedure.

Experience with Alemtuzumab (Campath-1H) as induction agent in renal transplantation followed by steroid-free immunosuppression.

BACKGROUND: The purpose of this study was to describe the initial experience with alemtuzumab as induction followed by steroid-free immunosuppression in kidney transplantation. METHODS: One hundred patients who received renal transplants from living and deceased donors were followed for a median period of 12 months (range = 1 to 12). A 30-mg intravenous dose of Alemtuzumab was administered on the transplant day, preceded by a 500-mg methylprednisolone dose. Maintenance immunosuppression consisted in the use of a calcineurin inhibitor in association with mycophenolic acid. Maintenance C2 levels of cyclosporine were between 400 and 600 ng/dL; or of tacrolimus, between 4 and 7 ng/dL. Prophylaxis included valgancyclovir, trimethoprim-sulfamethoxasole, and nystatin. All patients were evaluated for acute rejection episodes, adverse events, or death. RESULTS: The cumulative incidences of acute rejection at 1, 3, 6, and 12 months were 0%, 4% (n = 4), 5% (n = 5), and 8% (n = 8), respectively. Most episodes were Banff 1 a or b (88%). The infectious complication rate was 23%. There was no case of cytomegalovirus infection or posttransplant lymphoproliferative disease. Three patients died: one due to tuberculosis; one, sepsis; and one, an acute coronary event. No patient was lost to follow-up. CONCLUSIONS: This study suggested the safety and efficacy of Campath-1H as an induction agent in renal transplant recipients.

Conversion therapy to everolimus in renal transplant recipients: results after one year.

BACKGROUND: Two new diagnoses have been causing graft loss during long-term follow-up, namely, chronic nephropathy and anticalcineurinic toxicity. The advent of the mammalian target of rapamycin (m-TOR) obviates anticalcineurine toxicity and reduces posttransplant malignancy incidence with good inmunosuppressive potential. We examinated the renal and metabolic behavior in renal transplant recipients who required conversion from an anticalcineurinic (cyclosporine or tacrolimus) to an m-TOR inhibitor (everolimus) as part of their immunosuppressive maintenance therapy. MATERIALS AND METHODS: Twenty-one first renal transplant recipients had everolimus added to their inmunosuppressive therapy combined with an antimetabolite (mycophenolate mofetil or sodium mycophenolate). The mean age of the patients was 35 +/- 17 years (range, 6 to 65). The prevalence of male recipients was 57%; the overall mean weight, 64 kg (range, 48 to 95). All patients were hispanic with 15 transplants from cadaveric donors (71%). The mean follow-up posttransplant was 18 months (range, 3 to 40) and the mean follow-up on everolimus, 10 months (range, 2 to 22). RESULTS: There was no mortality or graft loss, but there were 3 (17%) biopsy-confirmed acute rejection episodes. There were no significant changes in metabolic function pre- or postconversion. Regarding renal function, the mean creatinine serum showed a trend to decline: preconversion 1.7 mg/dL; postconversion 1.5 mg/dL. In 10 patients, it was possible to discontinue at least one antihypertensive medication (48%). CONCLUSIONS: Everolimus was an effective medication to manage renal transplant patients. It produced metabolic stability and low myelotoxicity, despite combination with an antimetabolite (mycophenolic acid). Also, reduction of antihypertensive medications was an additional benefits for many patients.

Aeromonas aquariorum sp. nov., isolated from aquaria of ornamental fish.

During a survey to determine the prevalence of Aeromonas strains in water and skin of imported ornamental fish, 48 strains presumptively identified as Aeromonas were isolated but they could not be identified as members of any previously described Aeromonas species. These strains were subjected to a polyphasic approach including phylogenetic analysis derived from gyrB, rpoD and 16S rRNA gene sequencing, DNA-DNA hybridization, MALDI-TOF MS analysis, genotyping by RAPD and extensive biochemical and antibiotic susceptibility tests in order to determine their taxonomic position. Based on the results of the phylogenetic analyses and DNA-DNA hybridization data, we describe a novel species of the genus Aeromonas, for which the name Aeromonas aquariorum sp. nov. is proposed, with strain MDC47T (=DSM 18362T =CECT 7289T) as the type strain. This is the first Aeromonas species description based on isolations from ornamental fish.

Limited efficacy of a nonrestricted intervention on antimicrobial prescription of commonly used antibiotics in the hospital setting: results of a randomized controlled trial.

Most interventions aimed at diminishing the use of antimicrobials in hospitals have focussed on newly introduced antibiotics and very few have been randomly controlled. We evaluated the impact on antibiotic consumption of an intervention without restrictions in antibiotic use, focussed on commonly used antibiotics with a controlled randomized trial. All new prescriptions of levofloxacin, carbapenems, or vancomycin in hospitalized patients were randomized to an intervention or a control group. Intervention consisted of an antibiotic regimen counselling targeted to match local antibiotic guidelines, performed using only patients' charts. Clinical charts of patients assigned to the control group were reviewed daily by a pharmacist. The primary endpoint was a reduction in consumption of the targeted antibiotics. Two hundred seventy-eight prescriptions corresponding to 253 patients were included: 146 were assigned to the intervention and 132 to the control group. Total consumption of the targeted antibiotics (median [IQR]) was slightly lower in the intervention (8 [4-12] defined daily doses [DDDs] per patient) than in the control group (10 [6-16] DDDs per patient; p = 0.04). No differences in number of DDDs were observed when antibiotics of substitution were included (11.05 [6-18.2] vs 10 [6-16.5] in the intervention and control groups, respectively, p = 0.13). The total number of days on treatment with the targeted antibiotics was lower in the intervention (4 [3-7] days per patient) than in the control group (6 [4-10] days per patient; p = 0.002). Differences in number of days on treatment only reached statistical significance in the prescriptions of carbapenems. There were no differences between intervention and control groups in terms of number of deaths, hospital readmissions, length of hospital stay, or antibiotic costs. In this trial, an intervention without restrictions focussed on antimicrobial prescriptions of commonly used antibiotics in the hospital setting had a limited efficacy to reduce consumption and did not save costs. Future strategies to promote a more rational antimicrobial use should be evaluated with a randomized controlled design.

Luxación recidivante de prótesis total de cadera / Recurrent dislocation in total hip replacement

Más del 50% de los casos de caderas inestables se pueden resolver mediante la reducción cerrada de la luxación. Un intento sistemático para entender la causa de la inestabilidad, normalmente suele conducir a un tratamiento eficaz del problema cuando hace falta cirugía. Los componentes constreñidos o de retención han mejorado las posibilidades de resolver los problemas difíciles de inestabilidad, pero los efectos negativos potenciales de estos componentes deben tenerse también en cuenta. Los implantes tripolares, cuyos resultados clínicos en la literatura son muy alentadores, ocupan un lugar de elección en el arsenal terapéutico de la inestabilidad protética

Over 50% of unstable hip cases can be solved by carrying out a closed reduction of the dislocation. A systematic attempt to understand the cause of the instability, normally results in an efficient treatment of the condition in cases where surgery is required. Constrained ­ also known as retentive ­ components have enhanced the possibilities of addressing difficult instability problems, although the potential negative effects of these implants also need to be considered. Tripolar implants, whose clinical results are very encouraging according to the literature, should occupy a prominent place among the therapeutic options available for prosthetic instability

Updated phylogeny of the genus Aeromonas.

Recent phylogenetic studies of the genus Aeromonas based on gyrB and rpoD gene sequences have improved the phylogeny based on 16S rRNA gene sequences first published in 1992, particularly in the ability to split closely related species. These studies did not include the recently described species Aeromonas simiae and Aeromonas molluscorum and only a single strain of Aeromonas culicicola was available for analysis at that time. In the present work, these Aeromonas species and newly isolated strains of A. culicicola were examined. Sequence analysis indicates that A. simiae and A. molluscorum belong to non-described phylogenetic lines of descent within this genus, which supports the original description of both species. The most closely related species are Aeromonas schubertii and Aeromonas encheleia, respectively, which is consistent with 16S rRNA gene sequencing results. However, while the five strains of A. molluscorum showed nucleotide differences in their gyrB and rpoD gene sequences, the only two known A. simiae strains exhibited identical gene sequences, suggesting that they are isolates of the same strain. On the basis of the rpoD gene sequence phylogeny, A. culicicola strains from the original description and new isolates from drinking water and ornamental fish clustered within the species Aeromonas veronii, suggesting inconsistencies with previous results. Other strains with previously controversial taxonomy and new isolates from other studies were included in this study in order to clarify their phylogenetic affiliation at the species level.

First record of the rare species Aeromonas culicicola from a drinking water supply.

We describe the recovery of the rare species Aeromonas culicicola, so far known only in mosquitoes in India, from a drinking water supply in Spain. Typing, using enterobacterial repetitive intergenic consensus-PCR, revealed that the 27 new isolates belonged to 3 very closely related strains. These strains were genetically identified by 16S rRNA gene sequencing. Spanish strains differed from the mosquito strains in three nucleotide positions. The AHCYTOEN gene was present in these water strains, which may have a public health significance.

Phylogenetic analysis of the genus Aeromonas based on two housekeeping genes.

The phylogenetic relationships of all known species of the genus Aeromonas, and especially Aeromonas bestiarum and Aeromonas salmonicida, were investigated on 70 strains using the rpoD sequence, which encodes the sigma70 factor. This analysis was complemented with the sequence of gyrB, which has already proven useful for determining the phylogenetic relationships in the genus. Nucleotide sequences of rpoD and gyrB showed that both genes had similar substitution rates (< 2 %) and a similar number of variable positions (34 % for rpoD versus 32 % for gyrB). Strain groupings by analysis of rpoD, gyrB and a combination of both genes were consistent with the taxonomic organization of all Aeromonas species described to date. However, the simultaneous analysis of both clocks improved the reliability and the power to differentiate, in particular, closely related taxa. At the inter-species level, gyrB showed a better resolution for differentiating Aeromonas sp. HG11/Aeromonas encheleia and Aeromonas veronii/Aeromonas culicicola/Aeromonas allosaccharophila, while rpoD more clearly differentiated A. salmonicida from A. bestiarum. The analysis of rpoD provided initial evidence for clear phylogenetic divergence between the latter two species.