Rinderpest eradication: challenges for remaining disease free and implications for future eradication efforts.

In 2011, the World Organisation for Animal Health (OIE) and the Food and Agriculture Organization of the United Nations (FAO) declared global freedom from rinderpest, formally announcing that rinderpest virus infections had been eliminated from susceptible livestock populations. At the same time, it was recognised that rinderpest virus, and material containing rinderpest virus, remained stored in an unspecified number of facilities across the world. Although natural infections had been eliminated, there remained a risk that rinderpest could reoccur if such infectious material accidentally leaked or was intentionally released from one of these facilities into a susceptible animal population. To minimise this risk, the OIE and FAO, with the support of international partners, set in place a framework to: reduce the quantity of remaining rinderpest-virus-containing material; ensure that such material was only stored in high-security facilities; regulate any handling or manipulation of the virus; maintain vigilance amongst livestock keepers and Veterinary Services in the post-eradication era; and develop contingency plans to deal with any suspected or actual reoccurrence of rinderpest disease. In 2016, five years after the declaration of global freedom from rinderpest, official reports to the OIE show that virus and virus-containing material remain stored in 21 countries worldwide in 22 separate facilities, of which only five have been inspected and approved for holding rinderpest virus or vaccine. There is still much work to be done to further reduce the risk of a reoccurrence.

En 2011, l'Organisation mondiale de la santé animale (OIE) et l'Organisation des Nations Unies pour l'alimentation et l'agriculture (FAO) ont annoncé officiellement l'élimination de l'infection due au virus de la peste bovine dans les populations d'animaux d'élevage sensibles, déclarant ainsi la planète indemne de cette maladie. Parallèlement, les deux organisations faisaient état de l'existence d'un nombre indéterminé d'établissements dans le monde détenant des stocks du virus bovipestique ainsi que des produits contenant ce virus. Malgré l'élimination de l'infection chez ses hôtes naturels, un risque de réapparition de la peste bovine subsiste en cas de fuite accidentelle ou d'émission délibérée de ces produits infectieux dans les populations animales sensibles à partir de l'un de ces établissements. Afin de minimiser ce risque, l'OIE et la FAO soutenus par leurs partenaires internationaux ont mis en place un cadre visant plusieurs objectifs : réduire les quantités restantes de produits contenant le virus de la peste bovine dans le monde ; veiller à ce que ces produits ne soient stockés que dans des établissements de haute sécurité ; réglementer les conditions de détention et de manipulation du virus ; poursuivre la surveillance exercée par les éleveurs et les Services vétérinaires au cours de la phase post-éradication ; concevoir des plans d'urgence visant à faire face à toute réapparition suspectée ou confirmée de la peste bovine. En 2016, soit cinq ans après la déclaration de l'éradication mondiale de la peste bovine, il ressort des rapports officiels adressés à l'OIE que 21 pays détiennent encore des stocks du virus de la peste bovine ou des produits contenant ce virus, répartis en 22 établissements distincts dont seulement cinq ont fait l'objet d'une inspection et ont été dûment habilités à détenir des stocks de virus de la peste bovine ou de vaccins contre cette maladie. Il reste donc encore beaucoup à faire pour continuer à réduire le risque de réapparition de la peste bovine.

En 2011, la Organización Mundial de Sanidad Animal (OIE) y la Organización de las Naciones Unidas para la Alimentación y la Agricultura (FAO) anunciaron oficialmente que las infecciones causadas por el virus de la peste bovina habían sido eliminadas de las poblaciones sensibles de ganado, declarando así que el mundo quedaba libre de la enfermedad. Al mismo tiempo, significaron que un número no especificado de instalaciones dispersas por el mundo albergaban muestras del virus y otros productos que lo contenían. Aunque las infecciones naturales habían quedado eliminadas, subsistía el riesgo de reaparición de la peste bovina si en una de esas instalaciones se producía una fuga accidental o una liberación intencionada de material infeccioso y este entraba en contacto con una población animal sensible. Para reducir al mínimo tal riesgo, la OIE y la FAO, con apoyo de colaboradores internacionales, definieron un dispositivo encaminado a: reducir el volumen de material restante con contenido viral de la peste bovina; garantizar que ese material fuera conservado únicamente en instalaciones de alta seguridad; reglamentar toda manipulación del virus; mantener la vigilancia entre cuidadores de ganado y Servicios Veterinarios en el periodo posterior a la erradicación; y elaborar planes de emergencia para responder a toda reaparición, presunta o confirmada, de la peste bovina. En 2016, cinco años después de la declaración de ausencia mundial de peste bovina, los informes oficiales remitidos a la OIE daban fe de que había virus y productos que lo contenían en 22 instalaciones situadas en 21 países del mundo, de las que solo cinco habían sido inspeccionadas y homologadas para albergar virus de la peste bovina o vacunas contra la enfermedad. Queda pues mucho trabajo por delante para reducir en mayor medida el riesgo de reaparición.

Early thromboelastometry variables predict maximum clot firmness in children undergoing cardiac and non-cardiac surgery.

BACKGROUND: Early clot amplitudes measured on thromboelastometry (ROTEM®) predict maximum clot firmness (MCF) in adults. In this multicentre, retrospective study, we aimed to confirm the suspected relationship between early ROTEM® variables and MCF, in children undergoing cardiac or non-cardiac surgery. METHODS: 4762 ROTEM® tests (e.g. EXTEM, INTEM, FIBTEM, APTEM, and HEPTEM) performed in children undergoing cardiac or non-cardiac surgery at three University hospitals between January 2011 and June 2014 were reviewed. To assess the correlation between clot amplitudes measured after 5, 10 and 15 min and MCF, each variable was compared with the corresponding MCF by calculating Spearman's correlation coefficient. RESULTS: For the EXTEM® test, we observed that amplitude measured after 5 min (A5: r=0.91, P<0.001), 10 min (A10: r=0.95, P<0.001) and 15 min (A15: r=0.96, P<0.001) were strongly correlated to MCF. The same correlations were observed for INTEM® test (A5: r=0.93, P<0.001; A10: r=0.97, P<0.001; A15: r=0.97, P<0.001), and FIBTEM® test (A5: r=0.93, P<0.001; A10: r=0.94, P<0.001; A15: r=0.96, P<0.001). In addition, the amplitudes measured after five, 10 and 15 min were also strongly correlated with MCF in the APTEM® and the HEPTEM® tests. Receiver operating characteristics (ROC) analysis confirmed that A5, A10, A15 strongly predicted decreased MCF on all ROTEM® tests. CONCLUSIONS: This study confirmed that early values of clot amplitudes measured as soon as five, 10 or 15 min after clotting time could be used to predict maximum clot firmness in all ROTEM® tests.

The management of paediatric acute pain in Spain in 2021: Results of a national survey among paediatric anaesthesiologists.

OBJECTIVE: To improve knowledge about routine clinical practice in the management of paediatric acute pain in Spain. METHODS: A telematic survey was conducted via the Internet on a representative sample of healthcare professionals involved in the management of paediatric acute pain (specifically anaesthesiologists) in Spain. The survey included 28 questions about their usual clinical practice in the assessment and treatment of acute pain, and also training and organisational aspects in paediatric acute pain. RESULTS: The survey was completed during March 2021 by 150 specialists in anaesthesiology. The respondents widely experienced in the management of acute paediatric pain (mean years of experience: 14.3: SD: 7.8), essentially in acute postoperative pain (97% of cases). Although 80% routinely used validated paediatric acute pain assessment scales, only 2.6% used specific scales adapted for patients with cognitive impairment. Most of the respondents routinely used analgesic drugs such as paracetamol (99%) or metamizole (92%), but only 84% complemented these drugs with a loco-regional blocking technique or other non-steroidal anti-inflammatory drugs (62%). Furthermore, only 62.7% acknowledged having received specific training in paediatric acute pain, only 45% followed hospital institutional protocols, and a scant 28% did so through paediatric pain units. CONCLUSIONS: The survey identified important points for improvement in the training and organisation of acute pain management in Spanish paediatric patients.

Dugbe virus ovarian tumour domain interferes with ubiquitin/ISG15-regulated innate immune cell signalling.

The ovarian tumour (OTU) domain of the nairovirus L protein has been shown to remove ubiquitin and interferon-stimulated gene 15 protein (ISG15) from host cell proteins, which is expected to have multiple effects on cell signalling pathways. We have confirmed that the OTU domain from the L protein of the apathogenic nairovirus Dugbe virus has deubiquitinating and deISGylating activity and shown that, when expressed in cells, it is highly effective at blocking the TNF-α/NF-κB and interferon/JAK/STAT signalling pathways even at low doses. Point mutations of the catalytic site of the OTU [C40A, H151A and a double mutant] both abolished the ability of the OTU domain to deubiquitinate and deISGylate proteins and greatly reduced its effect on cell signalling pathways, confirming that it is this enzymic activity that is responsible for blocking the two signalling pathways. Expression of the inactive mutants at high levels could still block signalling, suggesting that the viral OTU can still bind to its substrate even when mutated at its catalytic site. The nairovirus L protein is a very large protein that is normally confined to the cytoplasm, where the virus replicates. When the OTU domain was prevented from entering the nucleus by expressing it as part of the N-terminal 205 kDa of the viral L protein, it continued to block type I interferon signalling, but no longer blocked the TNF-α-induced activation of NF-κB.

[Retrospective descriptive study about the use of levosimendan in children undergoing surgical correction for congenital heart disease]. / Estudio descriptivo retrospectivo sobre la utilización de levosimendán en niños sometidos a corrección quirúrgica de cardiopatía congénita.

OBJECTIVES: To describe the use of levosimendan for compassionate use in children undergoing surgery for congenital heart disease, as well as survival rates, and the variations in the haemodynamic and analytical variables studied. MATERIAL AND METHODS: An observational retrospective descriptive study was performed, using a review of clinical histories, from May 2005 to January 2010. Haemodynamic and analytical variables pre- and post- levosimendan administration, drugs used, and their dosages, and any adverse reactions were recorded. RESULTS: Forty two children, 38 of them undergoing surgical correction, between the ages of four days and 5.75 years (median 92 days) were included. The drug was infused on 46 occasions. Four children received two doses. The infusion rate was among 0.1 to 0.6 µg × kg⁻¹ × min⁻¹. Only one patient received a loading dose. In 15 administrations (32.6%), the same dose was maintained throughout the infusion period. In 19 cases (41.3%), the dose was increased or decreased according to the need for vasoactive support. In surgical patients, overall survival after 30 days of the administration, calculated using the Kaplan-Meier method, was 80%. Blood lactate levels were statistically associated with mortality (P<.001). CONCLUSIONS: There were no uniform criteria for using levosimendan, and it was only used as a rescue drug. Overall survival was similar to that reported in adult clinical trials. Clinical trials also need to be carried out in paediatric patients to determine the role of levosimendan in surgical practice, in order to develop and establish a clinical protocol for its use in children.

Peste des Petits Ruminants Virus.

Peste des petits ruminants virus (PPRV) causes a severe contagious disease of sheep and goats and has spread extensively through the developing world. Because of its disproportionately large impact on the livelihoods of low-income livestock keepers, and the availability of effective vaccines and good diagnostics, the virus is being targeted for global control and eventual eradication. In this review we examine the origin of the virus and its current distribution, and the factors that have led international organizations to conclude that it is eradicable. We also review recent progress in the molecular and cellular biology of the virus and consider areas where further research is required to support the efforts being made by national, regional, and international bodies to tackle this growing threat.

The synthesis and turnover of 5'-nucleotidase in primary cultured hepatocytes.

The synthesis and degradation of 5'-nucleotidase has been studied in rat hepatocytes. Primary cultures of rat hepatocytes were established with the cells showing evidence of polarity after 24-36 h in culture. After a 30 h lag period 5'-nucleotidase activity increased to a plateau level similar to the activity found in whole liver. The half life of the enzyme after reaching the plateau of activity was 22.8 h. Pulse-chase biosynthetic labelling studies of 5'-nucleotidase in the cultured hepatocytes using [35S]methionine showed that the 5'-nucleotidase monomer was synthesised as an Mr 67,000 form which was converted to the mature Mr 72,000 form. [35S]Methionine labelling studies in the presence of tunicamycin showed that the unglycosylated protein monomer was an Mr 57,000 form. The immature Mr 67,000 form of 5'-nucleotidase was sensitive to endoglycosidase H, whereas the mature form was sensitive only to endoglycosidase F. The data presented are consistent with 5'-nucleotidase in a polarised cell being synthesised and processed like other membrane glycoproteins, in contrast to earlier reports.

Improved technique for transient expression and negative strand virus rescue using fowlpox T7 recombinant virus in mammalian cells.

The suitability of recombinant T7 polymerase produced using either the highly attenuated MVA strain of vaccinia (MVA-T7) or fowlpox virus (FP-T7) for transient expression and negative strand virus rescue was compared in two mammalian cell lines (MDBK and Vero) and in primary cells of bovine, ovine and caprine origin. Such primary cells are more permissive for the growth of wild type strains of morbilliviruses, such as Rinderpest virus and Peste des petits ruminants virus. MVA-T7 was found to be highly cytopathic in the primary cells, multiplying rapidly and killing the cells within 3-5 days of infection, even when very low multiplicities of infection (MOI) were used. In contrast, FP-T7, which appeared to express similar amounts of T7 polymerase, was found to be non-cytopathic in a variety of primary and established cell lines of mammalian origin and was suitable for use in virus rescue experiments. MDBK cells and primary cells, unlike Vero cells, could not be efficiently transfected and so were unsuitable for virus rescue. Optimal conditions for rinderpest virus rescue in Vero cells were established using FP-T7 in place of MVA-T7. This system will be suitable for rescuing other viruses which grow in Vero cells.

The sequence of the N and L genes of rinderpest virus, and the 5' and 3' extra-genic sequences: the completion of the genome sequence of the virus.

We have sequenced the nucleocapsid (N) and polymerase (L) genes of the vaccine strain of rinderpest, and the 5' and 3' terminal domains of the genome. Together with previously published data, this completes the sequence of the entire genome of rinderpest virus. The L gene is identical in length to that of measles virus, encoding a 2183 amino acid protein with a calculated molecular weight of 248,100. The L protein sequence of three morbilliviruses is highly conserved, greater than 76% of residues being identical or conserved in all sequences. The N protein was, as for other sequenced genes, essentially identical to that of the virulent parent. The viral genome is 15,881 bases in length, similar to that of measles virus and slightly longer than that of canine distemper virus. The terminal sequences of the genome and those at the gene boundaries were compared to the analogous regions of other morbilliviruses and representatives of related groups of paramyxoviruses.

Editing of morbillivirus P gene transcripts in infected animals.

RNA editing in the Morbillivirus genus in vivo was investigated by applying a polymerase chain reaction-based primer extension technique to measure the edited and non-edited mRNA transcripts. In this genus of the Paramyxoviridae the P gene transcript is altered by the co-transcriptional addition of one extra G residue to produce the mRNA for the V non-structural protein. Using tissues of phocine distemper virus (PDV) infected seals, canine distemper virus (CDV) infected dogs and rinderpest virus (RPV) infected cattle, it was demonstrated that editing occurs in vivo. The P:V mRNA ratios were generally similar to those found in tissue culture infections with the same virus and a minor fraction of transcripts had 2-4 extra G residues. In one seal brain infected with PDV the ratio of P:V mRNA was reversed but no differences were found in the levels of mRNA editing in different tissues from the same animal infected with CDV or RPV. However, variation was seen between animals infected with different isolates of RPV and even between animals infected with the same isolate of RPV.

Characterization of two insulin-binding components of rat-liver plasma membranes.

We have identified and isolated two forms of insulin receptor from rat-liver plasma membranes. The smaller (Mr = 90k) is a single polypeptide. The same polypeptide appears to be the insulin-binding site of the larger Mr = 280k). Only the larger, multisubunit, receptor shows high-affinity binding of insulin and negative cooperativity in its dissociation kinetics.

Structural analysis of homologous repeated domains in alpha-actinin and spectrin.

The amino acid sequences of chick and slime mould alpha-actinin each contain four repeats of approximately 122 residues. These repeats are homologous to the 18-22 repeats, each of approximately 106 residues, found in the alpha and beta subunits of spectrin and fodrin, and to the multiple repeats of approximately 110 residues found in the Duchenne muscular dystrophy protein (dystrophin). The repeats correspond to the elongated rod-like portion of these molecules. We present a multiple sequence alignment of 21 repeats from this superfamily (8 alpha-actinin and 13 spectrin/fodrin), based on optimal pairwise alignments, from which a characteristic consensus pattern of amino acid types is deduced. Trp 46 is invariant in all but one repeat, and physicochemical classes of amino acids are conserved at 25 other positions. Secondary structure prediction on both the alpha-actinin and spectrin repeats taken together with the distribution of proline residues in the sequences, strongly suggest that each repeated domain consists of a four-helix structure. Our predictions differ significantly from previous three-helix models based on analyses of fewer sequences. To determine possible interdomain regions, sites of limited proteolysis of the native chick alpha-actinin dimer were determined and located in the amino acid sequence. The majority of these sites were in corresponding positions in different repeats within a segment predicted as a long helix. We propose a model, consistent with the overall dimensions of the rod-like portions of the molecules, in which these long, probably interrupted helices, link adjacent domains.

Development of new generation rinderpest vaccines.

Veterinary science has benefited much from the advances in biotechnology during the past 20 years. New and improved diagnostic techniques for infectious diseases have been developed and new and highly effective vaccines to prevent such diseases have been introduced and more have been, or are about to be, field-tested. The latest development in negative strand virology, reverse genetics, the ability to rescue live virus from a DNA copy of the RNA genome, is being used to address questions concerning virus pathogenicity at the molecular level and to produce "marker" vaccines, i.e. vaccines that allow serological identification of all vaccinated animals. Such a vaccine would greatly benefit the continuing campaign for the global eradication of rinderpest since it would then be possible, by serological means, to detect wild type virus circulating in local areas or regions where it is still necessary to vaccinate and where the vaccination levels are below those required to eliminate the virus. Here we describe different approaches we have taken to produce such a vaccine using reverse genetics to add a marker to the existing and widely used Plowright rinderpest vaccine.

[Effect of several inhalation anesthetics on ectopic cardiac automaticity. Intervention of the calcium ion]. / Efecto de diferentes anestésicos inhalatorios sobre el automatismo cardíaco ectópico. Intervención del ion calcio.

The effect of halothane, enflurane and isoflurane (at concentrations ranging from 0.1 v/v% to 5 v/v%) on ventricular automaticity induced by a local injury, has been studied in the isolated right ventricle of the rat. Both, halothane and isoflurane, effectively reduces ventricular frequency at all concentrations tested. On the contrary, enflurane (0.3, 0.5 and 1 v/v%) increases ventricular automaticity. The effect of enflurane was either potentiated or reduced respectively in the presence of lower or higher calcium concentrations.

[Analgesic effectiveness and repercussions on the progress of labor of small doses of bupivacaine and fentanyl in continuous peridural perfusion]. / Eficacia analgésica y repercusión sobre la evolución del parto de bajas dosis de bupivacaína y fentanilo en perfusión peridural continua.

OBJECTIVES: To compare the analgesic efficacy and repercussion on labor of early administration of two different concentrations of bupivacaine/fentanyl in continuous epidural perfusion, in comparison with a control group receiving no epidural anesthesia. PATIENTS AND METHODS: One hundred fifty patients were distributed among 3 groups. Group I (n = 50) received no epidural analgesia. Group II (n = 50) and III (n = 50) received test doses of 3 ml of bupivacaine plus adrenalin 1/200,000. After 5 minutes each patient in the study groups received 13 ml of the solution assigned (group II: 0.04% bupivacaine plus adrenalin 1/2,500,000 and fentanyl 2.5 micrograms/ml; group III: 0.0625% bupivacaine plus adrenalin 1/1,600,000 and fentanyl 2 micrograms/ml). Five minutes later a perfusion of 12 ml/h-1 of the same solution was delivered until dilation was complete. RESULTS: Epidural perfusion was started at 2.5 +/- 0.93 cm of dilation (group II) and 2.3 +/- 0.92 cm (group III). There were no statistically significant differences in either duration of labor until full dilation or expulsion among the groups. Pain assessed on a visual analog scale evolved from a baseline mean of 4.5 to 5 in the three groups, reaching 8.9 +/- 0.74 (group I), 0.24 +/- 0.89 (group II) and 0.28 +/- 0.57 (group III). There were no significant differences in fetal presentation or Apgar scores among the three groups at the end of delivery. CONCLUSION: Both solutions provide good analgesia during labor with minimum undesirable side effects. Low epidural doses of bupivacaine and fentanyl started early do not affect the course of labor.

[Post-surgical paediatric pain: Nursing- PCA vs continuous I.V. infusion of tramadol ]. / Dolor postoperatorio en lactantes y niños pequeños: nursing-PCA vs perfusión i.v. de tramadol.

AIM: To evaluate the efficiency in the control of the post-surgical paediatric pain of the combination of a weak opioid [tramadol (T)] and an NSAID (paracetamol), comparing its administration through "Nursing-PCA" (NCA) techniques or through continuous i.v. infusion. METHODS: The investigation has been carried out in 30 patients (mean 9.5 months) selected according to their foreseeable degree of moderate-hard pain. All of them consumed in the postoperative period: rectal paracetamol (20 mg/Kg) every 8 hours and tramadol in two groups. Group I: PCA pump with tramadol that was handled by the nurse. Initial dose: 0.5 mg/Kg NCA, bolus injection 0.3 mg/Kg with an interval of 10 minutes for security and a highest dose of 1.2 mg/Kg/4 h every 4 hours. Group II: continuous infusion i.v. of tramadol (6 mg/Kg/24 h). The pain was evaluated, as well as the sedative action, saturation oxygen, respiratory and hemodynamics parameters, adverse effects, and efficiency during the first 24 hours, as well as the number of total dose of drugs asked in the Nursing PCA group. RESULTS: Pain decreased in both groups. There were more sedative effects in group II and the total dose of tramadol was higher. There were no cases of respiratory depression. CONCLUSIONS: The combination of tramadol and paracetamol through "Nursing PCA" has turned out to be an efficient method in the treatment of the post-surgical pain in little children and those whose are in their lacteal period. It is a possible alternative of the continuous infusion of Morphine in these patients.

Development and testing of a field diagnostic assay for peste des petits ruminants virus.

We have developed an immunochromatographic test for the diagnosis of peste des petits ruminants (PPR) under field conditions. The diagnostic assay has been tested in the laboratory and also under field conditions in Ivory Coast, Pakistan, Ethiopia and Uganda. The test is carried out on a superficial swab sample (ocular or nasal) and showed a sensitivity of 84% relative to PCR. The specificity was 95% over all nasal and ocular samples. The test detected as little as 10(3) TCID50 (50% tissue culture infectious doses) of cell culture-grown virus, and detected virus isolates representing all four known genetic lineages of peste des petits ruminants virus. Virus could be detected in swabs from animals as early as 4 days post-infection, at a time when clinical signs were minimal. Feedback from field trials was uniformly positive, suggesting that this diagnostic tool may be useful for current efforts to control the spread of PPR.

Peste des petits ruminants: a suitable candidate for eradication?

This year will see the final announcement, accompanied by much justifiable celebration, of the eradication from the wild of rinderpest, the 'cattle plague' that has been with us for so many centuries. The only known rinderpest virus (RPV) remaining is in a relatively small number of laboratories around the world, and in the stockpiles of vaccine held on a precautionary basis. As we mark this achievement, only the second virus ever eradicated through human intervention, it seems a good time to look at rinderpest's less famous cousin, peste des petits ruminants ('the plague of small ruminants') and assess if it should, and could, also be targeted for global eradication.