New clinical trials regulation in Spain: analysis of royal decree 1090/2015.

The coming into force of Directive 2001/20/EC represented a step forward in harmonising clinical trial regulation in European countries, guaranteeing a uniform protection of subjects participating in clinical research across Europe. However, it led to a disproportionate increase in the bureaucratization, and thus, it became evident that procedures needed to be simplified without detriment to patient's safety. Thus, Regulation 536/2014, that repealed Directive 2001/20/EC, with the aim of decreasing the growing bureaucratization and stimulating clinical research in Europe, established simplified procedures, such as regulating a common procedure for authorising trials in Europe, the institution of strict assessment timelines, or the definition of new concepts, such as "low-intervention clinical trial". The legal form of a Regulation allowed the norm to be directly applied to Member States without the need for transposition. By means of the new Royal Decree, the national legislation is adapted to make the application of the regulation feasible and it allows the development of the aspects that the Regulation leaves to national legislation. Both documents seek to stimulate clinical research with medicinal products to foster knowledge, facilitate transparency, and reinforce subjects' safety. This will surely be the case, but with this revision, we will look at the novelties and key aspects that are most relevant to investigators and we will analyse the consequences for all parties involved in clinical research.

A large-scale evaluation of peptide vaccines against foot-and-mouth disease: lack of solid protection in cattle and isolation of escape mutants.

A large-scale vaccination experiment involving a total of 138 cattle was carried out to evaluate the potential of synthetic peptides as vaccines against foot-and-mouth disease. Four types of peptides representing sequences of foot-and-mouth disease virus (FMDV) C3 Argentina 85 were tested: A, which includes the G-H loop of capsid protein VP1 (site A); AT, in which a T-cell epitope has been added to site A; AC, composed of site A and the carboxy-terminal region of VP1 (site C); and ACT, in which the three previous capsid motifs are colinearly represented. Induction of neutralizing antibodies, lymphoproliferation in response to viral antigens, and protection against challenge with homologous infectious virus were examined. None of the tested peptides, at several doses and vaccination schedules, afforded protection above 40%. Protection showed limited correlation with serum neutralization activity and lymphoproliferation in response to whole virus. In 12 of 29 lesions from vaccinated cattle that were challenged with homologous virus, mutant FMDVs with amino acid substitutions at antigenic site A were identified. This finding suggests the rapid generation and selection of FMDV antigenic variants in vivo. In contrast with previous studies, this large-scale vaccination experiment with an important FMDV host reveals considerable difficulties for vaccines based on synthetic peptides to achieve the required levels of efficacy. Possible modifications of the vaccine formulations to increase protective activity are discussed.

Emerging foot-and-mouth disease virus variants with antigenically critical amino acid substitutions predicted by model studies using reference viruses.

One of the major obstacles to the design of effective antiviral vaccines is the frequent generation of antigenic viral variants in the field. The types of variants that will become dominant during disease outbreaks is often unpredictable. However, here we report the genetic and antigenic characterization of emerging foot-and-mouth disease virus (FMDV) variants with antigenically critical amino acid substitutions predicted by model studies using reference viruses and monoclonal antibodies. The new variants belong to serotype C and have caused a number of recent disease outbreaks in Argentina. The variants harbor antigenically drastic amino acid substitutions in each of the antigenic sites identified in FMDV. In particular, a substitution found at a major antigenic site (site A, the G-H loop of VP1) had been repeatedly selected in viruses resistant to neutralization by monoclonal and polyclonal antibodies. The association of critical amino acid replacements at predicted positions with new FMD outbreaks has a number of implications for FMD epidemiology and for the design of vaccines intended to control diseases caused by highly variable RNA viruses.

Molecular characterization of human immunodeficiency virus type 1 isolates from Venezuela.

Eight HIV-1 isolates from Venezuela have been characterized by nucleotide sequencing of the entire reverse transcriptase (RT)- and surface glycoprotein (gp 120)-coding regions. Average mutant frequencies were 2.5 x 10(-2) substitutions per nucleotide (s/nt) for the RT-coding region, and 10 x 10(-2) or 6.8 x 10(-2) s/nt for the gp120-coding region, depending on whether gaps introduced for optimal alignment were or were not, respectively, considered in the calculations. Phylogenetic trees were derived by maximum-likelihood, neighbor-joining, and maximum parsimony methods. In the trees derived from both RT- and gp120-coding regions, Venezuelan isolates cluster with subtype B viruses. However, the relative position of some of the isolates is considerably different in the two trees. Unique V3 loop amino acid sequences, not represented in the current database, have been identified among the Venezuelan isolates. In addition to representing the first molecular characterization of HIV-1 from Venezuela, the extensive genetic heterogeneity observed reinforces the interest in characterizing additional HIV-1 isolates worldwide for adequate vaccine design.

[Focal and segmental glomerulosclerosis in renal transplantation]. / Glomeruloesclerosis focal y segmentaria en el transplante renal.

In 179 patients subjected to 186 renal transplants, 30 renal biopsies were performed due to the presence of a proteinuria over 3.5 g/24 h or due to a reduction in glomerular filtration rate. Six of these biopsies, coming from 5 patients, disclosed morphological alterations compatible with focal segmental glomeruloesclerosis. Five of these were due to recurrence of the primary disease (in four patients) and in all, massive proteinuria appeared from 1 to 23 days after transplantation. Two patients with three transplants, evolved to renal failure and required dialysis in a period 12 months as a mean. The third patient, developed a nephrotic syndrome without renal failure and died 14 months after the renal transplant due to a stroke. In the fourth patient, the nephrotic syndrome disappeared 38 days after the transplant and remained with minimal proteinuria until his last follow up visit two years later. The primary disease of the fifth patient is unknown; the nephrotic syndrome appeared 68 months after the transplant and remitted spontaneously in 2 months. The renal biopsy showed focal and segmental lesions with partial effacement of epithelial foot processes. It is concluded that focal segmental glomerulosclerosis recurrence in renal transplant occurs with early massive proteinuria and frequently leads to renal failure and graft loss in no more than two years.

Muscular changes in Venezuelan wild horses naturally infected with Trypanosoma evansi.

Skeletal muscle biopsy specimens were taken from 10 male horses naturally infected with Trypanosoma evansi and from 10 uninfected males. An indirect fluorescent antibody test was used to provide a rapid and reliable indication of infection. Histological, histochemical and transmission electron microscopical techniques were used to examine skeletal muscle. The ultrastructural features in muscle fibres were those usually seen in autoimmune disease, namely fibre and capillary necrosis and mononuclear cell infiltration, consisting of macrophages. Changes in fibre-type percentages did not occur in response to the parasite, but alterations in fibre size and capillary supply were found in the parasitized horses.

Extensive antigenic diversification of foot-and-mouth disease virus by amino acid substitutions outside the major antigenic site.

The antigenic sites A and C (the G-H loop and the C terminus, respectively) in VP1 of foot-and-mouth disease virus (FMDV) have been considered the immunodominant regions of the virus involved in the induction of protection. Other antigenic sites have been described but their involvement in protection has not been established. Here we report that two closely related but serologically different FMDVs (the field isolate C3 Argentina/84 and the vaccine strain C3 Resende Br/55) have identical A and C sites but differ as other antigenic sites. Such differences have been documented by reactivity with a panel of 28 monoclonal antibodies (MAbs). The two viruses reacted to the same extent with each of 13 MAbs which recognized epitopes within sites A or C, but reacted differently with six out of 15 MAbs that recognized other sites. Accordingly, sequencing of the entire region coding for the capsid proteins, for both viruses, revealed four amino acid substitutions at three antigenic sites other than A and C. The results suggest that identity of sites A and C may not be sufficient to induce cross-protection, and provide the first evidence of significant antigenic diversification of FMDV in the field mediated by amino acid substitutions outside sites A or C.

Extensive antigenic heterogeneity of foot-and-mouth disease virus of serotype C.

The antigenic behavior of 46 field isolates of foot-and-mouth disease virus (FMDV) of serotype C has been studied with a panel of 24 monoclonal antibodies (MAbs) prepared against FMDV C1 or FMDV C3 Indaial. Reactivities were assayed by immunodot, immunoelectrotransfer blot, and neutralization of infectivity. The epitopes recognized by the 10 nonneutralizing MAbs are conserved in all isolates analyzed. In contrast, extreme antigenic heterogeneity is documented with regard to reactivity with 14 MAbs that, on this basis, define at least 12 epitopes involved in neutralization of FMDV of serotype C. The 31 isolates from South America were divided into 17 distinct antigenic groups and the 15 isolates from Europe into 7 groups. Lack of correspondence between antigenic composition and the origin--date and place of isolation--of the viruses was noted in several instances. Antigenic heterogeneity is shown among epidemiologically closely related FMDVs. In most--but not all--cases tested, a good correlation was found between binding of a neutralizing MAb to virions and its ability to neutralize infectivity. It is concluded that variation of epitopes involved in neutralization of FMDV is extensive among subtypes of serotype C and also among individual isolates of one subtype.

Ileo biliar. / Biliary ileum

Se presentan 11 casos de ileo biliar tratados en un periodo de 3 anos en una misma institucion. Representan el 4% de todas las obstrucciones intestinales y se hizo el diagnostico preoperatorio en 8 de los 11 enfermos. La baja morbilidad y la ausencia de mortalidad a pesar de ser enfermos ancianos, se debio a varios factores, como la preparacion con antibioticos, la precocidad de la cirugia, los cuidados intraoperatorios de asepsia y antisepsia y al criterio de solo extraer el calculo

Ileo biliar. / Biliary ileum

Se presentan 11 casos de ileo biliar tratados en un periodo de 3 anos en una misma institucion. Representan el 4% de todas las obstrucciones intestinales y se hizo el diagnostico preoperatorio en 8 de los 11 enfermos. La baja morbilidad y la ausencia de mortalidad a pesar de ser enfermos ancianos, se debio a varios factores, como la preparacion con antibioticos, la precocidad de la cirugia, los cuidados intraoperatorios de asepsia y antisepsia y al criterio de solo extraer el calculo