Adeno-Associated Virus Type 5 Infection via PDGFRα Is Associated With Interstitial Lung Disease in Systemic Sclerosis and Generates Composite Peptides and Epitopes Recognized by the Agonistic Immunoglobulins Present in Patients With Systemic Sclerosis.

OBJECTIVE: The etiopathogenesis of systemic sclerosis (SSc) is unknown. Platelet-derived growth factor receptors (PDGFRs) are overexpressed in patients with SSc. Because PDGFRα is targeted by the adeno-associated virus type 5 (AAV5), we investigated whether AAV5 forms a complex with PDGFRα exposing epitopes that may induce the immune responses to the virus-PDGFRα complex. METHODS: The binding of monomeric human PDGFRα to the AAV5 capsid was analyzed by in silico molecular docking, surface plasmon resonance (SPR), and genome editing of the PDGFRα locus. AAV5 was detected in SSc lungs by in situ hybridization, immunohistochemistry, confocal microscopy, and molecular analysis of bronchoalveolar lavage (BAL) fluid. Immune responses to AAV5 and PDGFRα were evaluated by SPR using SSc monoclonal anti-PDGFRα antibodies and immunoaffinity-purified anti-PDGFRα antibodies from sera of patients with SSc. RESULTS: AAV5 was detected in the BAL fluid of 41 of 66 patients with SSc with interstitial lung disease (62.1%) and in 17 of 66 controls (25.75%) (P < 0.001). In SSc lungs, AAV5 localized in type II pneumocytes and in interstitial cells. A molecular complex formed of spatially contiguous epitopes of the AAV5 capsid and of PDGFRα was identified and characterized. In silico molecular docking analysis and binding to the agonistic anti-PDGFRα antibodies identified spatially contiguous epitopes derived from PDGFRα and AAV5 that interacted with SSc agonistic antibodies to PDGFRα. These peptides were also able to bind total IgG isolated from patients with SSc, not from healthy controls. CONCLUSION: These data link AVV5 with the immune reactivity to endogenous antigens in SSc and provide a novel element in the pathogenesis of SSc.

Facial Affect Recognition by Patients with Schizophrenia Using Human Avatars.

People with schizophrenia have difficulty recognizing the emotions in the facial expressions of others, which affects their social interaction and functioning in the community. Static stimuli such as photographs have been used traditionally to examine deficiencies in the recognition of emotions in patients with schizophrenia, which has been criticized by some authors for lacking the dynamism that real facial stimuli have. With the aim of overcoming these drawbacks, in recent years, the creation and validation of virtual humans has been developed. This work presents the results of a study that evaluated facial recognition of emotions through a new set of dynamic virtual humans previously designed by the research team, in patients diagnosed of schizophrenia. The study included 56 stable patients, compared with 56 healthy controls. Our results showed that patients with schizophrenia present a deficit in facial affect recognition, compared to healthy controls (average hit rate 71.6% for patients vs 90.0% for controls). Facial expressions with greater dynamism (compared to less dynamic ones), as well as those presented from frontal view (compared to profile view) were better recognized in both groups. Regarding clinical and sociodemographic variables, the number of hospitalizations throughout life did not correlate with recognition rates. There was also no correlation between functioning or quality of life and recognition. A trend showed a reduction in the emotional recognition rate as a result of increases in Positive and Negative Syndrome Scale (PANSS), being statistically significant for negative PANSS. Patients presented a learning effect during the progression of the task, slightly greater in comparison to the control group. This finding is relevant when designing training interventions for people with schizophrenia. Maintaining the attention of patients and getting them to improve in the proposed tasks is a challenge for today's psychiatry.

Prevalence of plaque and dental decay in the first permanent molar in a school population of south Mexico City. / Prevalencia de placa dentobacteriana y caries dental en el primer molar permanente en una población escolar del sur de la Ciudad de México

Background: The first permanent molar is susceptible to acquire tooth decay since its eruption, due to its anatomy and because it has been exposed before other teeth. Method: An observational, prolective, transversal and comparative study in 194 students, with an average age of 9.9 ± 1.8 years. The evaluation of the dentobacterial plate (DBP) was analyzed using the O'Leary index and the tooth decay experience with the DMFS (sum of decayed, missing, extracted and filling dental surfaces) and DMFT (sum of decayed, missing, extracted and filling per tooth) indexes. Results: The prevalence of DBP in the first permanent molar was of 99.4% and tooth decay of 57.2%. The value of DMFT was 1.4 ± 1.4. The tooth decay experience was higher in children from 7.10 years old with a value of 2.2 ± 2.3, who are 7.9 times more likely to develop lesions than younger children (odds ratio: 8.9; 95% confidence interval: 4.1-19.5; p < 0.0001). We found an association between age and the values of the tooth decay experience indexes; even though these were weak in the case of DMF (r = 0.439), the model allowed to explain 19% of the association, and 22% for DMFT (r = 0.464). Conclusions: Tooth decay develops rapidly in the first permanent molars; however, it does not receive the necessary care because it is usually unknown that it is a permanent tooth.

Introducción: El primer molar permanente (PMP) es susceptible de caries a partir de su erupción por su anatomía y porque ha estado expuesto antes que otros dientes. Métodos: Estudio observacional, prolectivo, transversal, comparativo, en 194 escolares con una media de edad de 9.9 ± 1.8 años. La valoración de la placa dentobacteriana (PDB) se realizó con el índice de O'Leary, y la experiencia de caries con los índices CPOS (suma de superficies dentales cariadas, perdidas/extraídas y obturadas) y CPOD (suma de dientes cariados, perdidos/extraídos y obturados). Resultados: La prevalencia de PDB en el PMP fue del 99.4%, y la de caries, del 57.2%. El valor del CPOD fue de 1.4 ± 1.4. La experiencia de caries por superficie dental en los niños de 7.10 años de edad fue de 2.2 ± 2.3 y son 7.9 veces más susceptibles a desarrollar lesiones (razón de momios: 8.9; intervalo de confianza al 95%: 4.1-19.5; p < 0.0001) que los menores de esa edad. Se detectó asociación con la variable edad y los valores de los índices de experiencia de caries; aun cuando estas son débiles en el caso del CPOS (r = 0.439), el modelo permitió explicar el 19% de la asociación, y el 22% para el CPOD (r = 0.464). Conclusiones: La caries dental se desarrolla rápidamente en los PMP. En general, no se le da la importancia debida porque se desconoce que se presenta en un diente permanente.

Prevalencia de placa dentobacteriana y caries dental en el primer molar permanente en una población escolar del sur de la Ciudad de México / Prevalence of plaque and dental decay in the first permanent molar in a school population of south Mexico City

Resumen Introducción: El primer molar permanente (PMP) es susceptible de caries a partir de su erupción por su anatomía y porque ha estado expuesto antes que otros dientes. Métodos: Estudio observacional, prolectivo, transversal, comparativo, en 194 escolares con una media de edad de 9.9 ± 1.8 años. La valoración de la placa dentobacteriana (PDB) se realizó con el índice de O'Leary, y la experiencia de caries con los índices CPOS (suma de superficies dentales cariadas, perdidas/extraídas y obturadas) y CPOD (suma de dientes cariados, perdidos/extraídos y obturados). Resultados: La prevalencia de PDB en el PMP fue del 99.4%, y la de caries, del 57.2%. El valor del CPOD fue de 1.4 ± 1.4. La experiencia de caries por superficie dental en los niños de 7.10 años de edad fue de 2.2 ± 2.3 y son 7.9 veces más susceptibles a desarrollar lesiones (razón de momios: 8.9; intervalo de confianza al 95%: 4.1-19.5; p < 0.0001) que los menores de esa edad. Se detectó asociación con la variable edad y los valores de los índices de experiencia de caries; aun cuando estas son débiles en el caso del CPOS (r = 0.439), el modelo permitió explicar el 19% de la asociación, y el 22% para el CPOD (r = 0.464). Conclusiones: La caries dental se desarrolla rápidamente en los PMP. En general, no se le da la importancia debida porque se desconoce que se presenta en un diente permanente.

Abstract Background: The first permanent molar is susceptible to acquire tooth decay since its eruption, due to its anatomy and because it has been exposed before other teeth. Method: An observational, prolective, transversal and comparative study in 194 students, with an average age of 9.9 ± 1.8 years. The evaluation of the dentobacterial plate (DBP) was analyzed using the O'Leary index and the tooth decay experience with the DMFS (sum of decayed, missing, extracted and filling dental surfaces) and DMFT (sum of decayed, missing, extracted and filling per tooth) indexes. Results: The prevalence of DBP in the first permanent molar was of 99.4% and tooth decay of 57.2%. The value of DMFT was 1.4 ± 1.4. The tooth decay experience was higher in children from 7.10 years old with a value of 2.2 ± 2.3, who are 7.9 times more likely to develop lesions than younger children (odds ratio: 8.9; 95% confidence interval: 4.1-19.5; p < 0.0001). We found an association between age and the values of the tooth decay experience indexes; even though these were weak in the case of DMF (r = 0.439), the model allowed to explain 19% of the association, and 22% for DMFT (r = 0.464). Conclusions: Tooth decay develops rapidly in the first permanent molars; however, it does not receive the necessary care because it is usually unknown that it is a permanent tooth.

Variability of chlorophyll-a concentration in the Gulf of Guinea and its relation to physical oceanographic variables.

The Gulf of Guinea represents a wide tract of the African coast with complex and rich coastal ecosystems undergoing various pressures. The seasonal variations of chlorophyll-a concentration (Chla) along the Gulf of Guinea (GoG) and their relations with physical oceanographic variables were analyzed using satellite observations covering the period 2002-2012. The effects of sea surface temperature (SST), sea level anomalies (SLA), winds, geostrophic currents, eddy kinetic energy (EKE), mesoscale eddies and fronts were considered on a monthly time scale. The analysis for each unit area was carried out on a chlorophyll index (IChla) computed as the product of the mean distance from the coast to the eutrophic threshold (1 mg m-3 isoline) and the average Chla in the eutrophic area. The study, based on satellite-derived Chla, was allowed by the unprecedented coverage given by the products distributed by the ESA Ocean Colour Climate Change Initiative (OC_CCI) resulting from the merging of data from several satellite missions. The physical variables served as potential predictors in a statistical Boosted Regression Tree (BRT) model. To account for the heterogeneous nature of the GoG, the analysis was conducted on eight systems that made up a partition of the whole region defined on the basis of the BRT model results and climatological properties. The western-most domain, from Guinea-Bissau to Sierra Leone, was associated with upwelling properties in boreal winter and appeared to share some characteristics with the overall Northwest African upwelling system. The region of Ivory Coast and Ghana also had upwelling properties but the main upwelling season was in boreal summer. In general upwelling conditions with cold SST, negative SLA, fairly strong frontal activity, and moderate winds, appeared as the environmental window most favorable to high IChla values. For these systems, the BRT model fitted the IChla data well with a percentage of explained total deviance [Formula: see text] between 70% and 91% when using only physical oceanographic variables. Finally, the systems associated with the coasts of Nigeria to Gabon showed some mixed properties, with [Formula: see text] values of 54-60%. Among these systems, a common feature seemed to be the importance of river discharge to explain IChla variations. Where possible (for the Niger River in the Nigeria system), the addition of river data as predictor in the BRT model resulted in a significant increase of [Formula: see text] to 75%. Further progress is needed to understand the observed relationships and to predict how they can evolve in the face of climate change.

Correction to 'Food limitation of sea lion pups and the decline of forage off central and southern California'.

[This corrects the article DOI: 10.1098/rsos.150628.].

Food limitation of sea lion pups and the decline of forage off central and southern California.

California sea lions increased from approximately 50 000 to 340 000 animals in the last 40 years, and their pups are starving and stranding on beaches in southern California, raising questions about the adequacy of their food supply. We investigated whether the declining sea lion pup weight at San Miguel rookery was associated with changes in abundance and quality of sardine, anchovy, rockfish and market squid forage. In the last decade off central California, where breeding female sea lions from San Miguel rookery feed, sardine and anchovy greatly decreased in biomass, whereas market squid and rockfish abundance increased. Pup weights fell as forage food quality declined associated with changes in the relative abundances of forage species. A model explained 67% of the variance in pup weights using forage from central and southern California and 81% of the variance in pup weights using forage from the female sea lion foraging range. A shift from high to poor quality forage for breeding females results in food limitation of the pups, ultimately flooding animal rescue centres with starving sea lion pups. Our study is unusual in using a long-term, fishery-independent dataset to directly address an important consequence of forage decline on the productivity of a large marine predator. Whether forage declines are environmentally driven, are due to a combination of environmental drivers and fishing removals, or are due to density-dependent interactions between forage and sea lions is uncertain. However, declining forage abundance and quality was coherent over a large area (32.5-38° N) for a decade, suggesting that trends in forage are environmentally driven.

AAV Vectors Vaccines Against Infectious Diseases.

Since their discovery as a tool for gene transfer, vectors derived from the adeno-associated virus (AAV) have been used for gene therapy applications and attracted scientist to this field for their exceptional properties of efficiency of in vivo gene transfer and the level and duration of transgene expression. For many years, AAVs have been considered as low immunogenic vectors due to their ability to induce long-term expression of non-self-proteins in contrast to what has been observed with other viral vectors, such as adenovirus, for which strong immune responses against the same transgene products were documented. The perceived low immunogenicity likely explains why the use of AAV vectors for vaccination was not seriously considered before the early 2000s. Indeed, while analyses conducted using a variety of transgenes and animal species slowly changed the vision of immunological properties of AAVs, an increasing number of studies were also performed in the field of vaccination. Even if the comparison with other modes of vaccination was not systemically performed, the analyses conducted so far in the field of active immunotherapy strongly suggest that AAVs possess some interesting features to be used as tools to produce an efficient and sustained antibody response. In addition, recent studies also highlighted the potential of AAVs for passive immunotherapy. This review summarizes the main studies conducted to evaluate the potential of AAV vectors for vaccination against infectious agents and discusses their advantages and drawbacks. Altogether, the variety of studies conducted in this field contributes to the understanding of the immunological properties of this versatile virus and to the definition of its possible future applications.

Properties of the adeno-associated virus assembly-activating protein.

Adeno-associated virus (AAV) capsid assembly requires expression of the assembly-activating protein (AAP) together with capsid proteins VP1, VP2, and VP3. AAP is encoded by an alternative open reading frame of the cap gene. Sequence analysis and site-directed mutagenesis revealed that AAP contains two hydrophobic domains in the N-terminal part of the molecule that are essential for its assembly-promoting activity. Mutation of these sequences reduced the interaction of AAP with the capsid proteins. Deletions and a point mutation in the capsid protein C terminus also abolished capsid assembly and strongly reduced the interaction with AAP. Interpretation of these observations on a structural basis suggests an interaction of AAP with the VP C terminus, which forms the capsid protein interface at the 2-fold symmetry axis. This interpretation is supported by a decrease in the interaction of monoclonal antibody B1 with VP3 under nondenaturing conditions in the presence of AAP, indicative of steric hindrance of B1 binding to its C-terminal epitope by AAP. In addition, AAP forms high-molecular-weight oligomers and changes the conformation of nonassembled VP molecules as detected by conformation-sensitive monoclonal antibodies A20 and C37. Combined, these observations suggest a possible scaffolding activity of AAP in the AAV capsid assembly reaction.

[Eligibility of schizophrenia inpatients to participate in clinical trials]. / Elegibilidad de pacientes con esquizofrenia ingresados en una unidad de hospitalización psiquiátrica para participar en ensayos clínicos.

INTRODUCTION: This study assesses the potential eligibility of patients admitted to a psychiatric hospitalisation unit to take part in the major clinical trials based on schizophrenia treatment in clinical practice (CATIE, CUtLASS and EUFEST). MATERIAL AND METHODS: A retrospective evaluation by consulting the medical records of 241 subjects (59.8% males and 40.2% females, mean age 39.7±13.0 years), admitted consecutively over one year to psychiatric hospitalisation unit with a diagnosis of schizophrenia or another psychosis. The influence of the factors involved in the non-eligibility in each of the clinical trials is analysed using logistic regression analysis. RESULTS: Only 20.7%, 22.3%, and 22.5% of patients with schizophrenia or another psychosis would be eligible to participate in the CATIE, CUtLASS and EUFEST studies, respectively. The main factors involved in the non-eligibility were polytherapy with anti-psychotics (2 or more) (Odds Ratio (OR): 7.64, 95% confidence interval (CI): 3.06-19.06, P<.001), mental retardation (OR: 16.67, 95% CI: 1.75-166.67, P=.014), and resistance, intolerance or contraindication to any of the anti-psychotics of the study (OR: 3.68, 95% CI: 1.13-11.99, P=.030). CONCLUSIONS: Three out of every four patients with schizophrenia or another psychosis admitted to a psychiatric hospitalisation unit are not represented in the major clinical trials on schizophrenia treatment.

Development of AAVLP(HPV16/31L2) particles as broadly protective HPV vaccine candidate.

The human papillomavirus (HPV) minor capsid protein L2 is a promising candidate for a broadly protective HPV vaccine yet the titers obtained in most experimental systems are rather low. Here we examine the potential of empty AAV2 particles (AAVLPs), assembled from VP3 alone, for display of L2 epitopes to enhance their immunogenicity. Insertion of a neutralizing epitope (amino acids 17-36) from L2 of HPV16 and HPV31 into VP3 at positions 587 and 453, respectively, permitted assembly into empty AAV particles (AAVLP(HPV16/31L2)). Intramuscularly vaccination of mice and rabbits with AAVLP(HPV16/31L2)s in montanide adjuvant, induced high titers of HPV16 L2 antibodies as measured by ELISA. Sera obtained from animals vaccinated with the AAVLP(HPV16/31L2)s neutralized infections with several HPV types in a pseudovirion infection assay. Lyophilized AAVLP(HPV16/31L2) particles retained their immunogenicity upon reconstitution. Interestingly, vaccination of animals that were pre-immunized with AAV2--simulating the high prevalence of AAV2 antibodies in the population--even increased cross neutralization against HPV31, 45 and 58 types. Finally, passive transfer of rabbit antisera directed against AAVLP(HPV16/31L2)s protected naïve mice from vaginal challenge with HPV16 pseudovirions. In conclusion, AAVLP(HPV16/31L2) particles have the potential as a broadly protective vaccine candidate regardless of prior exposure to AAV.

Intranasal vaccination with AAV5 and 9 vectors against human papillomavirus type 16 in rhesus macaques.

Cervical cancer is the second most common cancer in women worldwide. Persistent high-risk human papillomavirus (HPV) infection has been identified as the causative event for the development of this type of cancer. Recombinant adeno-associated viruses (rAAVs) are currently being developed and evaluated as vaccine vector. In previous work, we demonstrated that rAAVs administered intranasally in mice induced high titers and long-lasting neutralizing antibodies against HPV type 16 (HPV16). To extend this approach to a more human-related species, we immunized rhesus macaques (Macaca mulatta) with AAVs expressing an HPV16 L1 protein using rAAV5 and 9 vectors in an intranasal prophylactic setting. An rAAV5-L1 vector followed by a boost with rAAV9-L1 induced higher titers of L1-specific serum antibodies than a single rAAV5-L1 immunization. L1-specific antibodies elicited by AAV9 vector neutralized HPV16 pseudovirions and persisted for at least 7 months post immunization. Interestingly, nasal application of rAAV9 was immunogenic even in the presence of high AAV9 antibody titers, allowing reimmunization with the same serotype without prevention of the transgene expression. Two of six animals did not respond to AAV-mediated intranasal vaccination, although they were not tolerant, as both developed antibodies after intramuscular vaccination with HPV16 virus-like particles. These data clearly show the efficacy of an intranasal immunization using rAAV9-L1 vectors without the need of an adjuvant. We conclude from our results that rAAV9 vector is a promising candidate for a noninvasive nasal vaccination strategy.

HPV16 E2 could act as down-regulator in cellular genes implicated in apoptosis, proliferation and cell differentiation.

BACKGROUND: Human Papillomavirus (HPV) E2 plays several important roles in the viral cycle, including the transcriptional regulation of the oncogenes E6 and E7, the regulation of the viral genome replication by its association with E1 helicase and participates in the viral genome segregation during mitosis by its association with the cellular protein Brd4. It has been shown that E2 protein can regulate negative or positively the activity of several cellular promoters, although the precise mechanism of this regulation is uncertain. In this work we constructed a recombinant adenoviral vector to overexpress HPV16 E2 and evaluated the global pattern of biological processes regulated by E2 using microarrays expression analysis. RESULTS: The gene expression profile was strongly modified in cells expressing HPV16 E2, finding 1048 down-regulated genes, and 581 up-regulated. The main cellular pathway modified was WNT since we found 28 genes down-regulated and 15 up-regulated. Interestingly, this pathway is a convergence point for regulating the expression of genes involved in several cellular processes, including apoptosis, proliferation and cell differentiation; MYCN, JAG1 and MAPK13 genes were selected to validate by RT-qPCR the microarray data as these genes in an altered level of expression, modify very important cellular processes. Additionally, we found that a large number of genes from pathways such as PDGF, angiogenesis and cytokines and chemokines mediated inflammation, were also modified in their expression. CONCLUSIONS: Our results demonstrate that HPV16 E2 has regulatory effects on cellular gene expression in HPV negative cells, independent of the other HPV proteins, and the gene profile observed indicates that these effects could be mediated by interactions with cellular proteins. The cellular processes affected suggest that E2 expression leads to the cells in to a convenient environment for a replicative cycle of the virus.

Human papillomavirus-specific immune therapy: failure and hope.

Recently, two prophylactic vaccines against the most significant oncogenic human papillomaviruses (HPV; 16 and 18) became available that efficiently protect against persistent HPV infection and cancer precursors. However, clinical trials performed with these vaccines did not provide evidence that they would influence the natural history of prevalent HPV infections, that is, their eventual malignant progression. Because, even under the optimistic assumption of high vaccine coverage, a significant reduction of cancer incidence can only be expected after two decades, there is a need for immune therapeutic strategies to be offered to persistently infected individuals who do not benefit from the prophylactic vaccines. Here, we describe the reasons for failure of most of the published approaches to HPV-specific therapies, highlight promising developments and present our view for future developments.

Combined prophylactic and therapeutic intranasal vaccination against human papillomavirus type-16 using different adeno-associated virus serotype vectors.

BACKGROUND: Cervical cancer is the second most frequent cancer among woman worldwide and is considered to be caused by infection with high-risk papilloma viruses. Genetic immunization using recombinant adeno-associated virus (rAAV) vectors has shown great promise for vaccination against human papillomavirus (HPV) infections. METHODS: rAAV5, -8 and -9 vectors expressing an HPV16 L1/E7 fusion gene were generated and applied intranasally for combined prophylactic and therapeutic vaccination of mice. RESULTS: The rAAV5 and the rAAV9 vectors showed efficient induction of both humoral and cellular immune responses, whereas rAAV8 failed to immunize mice by the intranasal route. The L1-specific immune response evoked by expression of the L1/E7 fusion gene, however, was lower than that evoked by expression of the L1 antigen alone. This deficiency could be compensated by application of Escherichia coli heat-labile enterotoxin or monophsphoryl lipid as adjuvant upon vaccination with rAAV5-L1/E7. Coimmunization of rAAV9-L1/E7 with rAAV5-L1 or boosting of rAAV9-L1/E7 with rAAV5-L1 strongly increased L1-specific neutralizing antibody titres to levels above those achieved by vaccination with vectors expressing L1 alone. Both vectors elicited a vibrant cytotoxic T-lymphocyte response against L1 or E7. Nasal immunization with rAAV5 or rAAV9 was superior to vaccination with HPV16-L1 virus-like particles (VLPs) or HPV16-L1/E7 CVLPs with respect to humoral and cellular immune responses. Vaccination with the rAAV vectors led to a significant protection of animals against a challenge with different HPV tumour cell lines. CONCLUSIONS: Our results show that rAAV5 and rAAV9 vectors are promising candidates for a non-invasive nasal vaccination strategy.

The therapeutic vaccine: is it feasible?

In contrast to the prophylactic HPV vaccines that exhibit great promise in reducing the burden of cervical cancer, there is limited progress towards the development of immune therapeutic strategies that would help those women who are already infected with high-risk HPV and do not benefit from the current vaccines. The reason for this drawback is the lack of knowledge about the immune mechanisms that control the growth of HPV-infected or -transformed cells in vivo. It became evident that the preclinical models in rodents provide only limited information about the performance of a candidate vaccine in humans. In particular, the immune correlate for a clinical response remains to be determined. On the other hand, HPV-related malignancies provide an excellent model for cancer immune therapies in general. There is hope that the continuous efforts of academic research combined with corporate involvement will finally present an efficient product.

Synthetic double-stranded RNAs are adjuvants for the induction of T helper 1 and humoral immune responses to human papillomavirus in rhesus macaques.

Toll-like receptor (TLR) ligands are being considered as adjuvants for the induction of antigen-specific immune responses, as in the design of vaccines. Polyriboinosinic-polyribocytoidylic acid (poly I:C), a synthetic double-stranded RNA (dsRNA), is recognized by TLR3 and other intracellular receptors. Poly ICLC is a poly I:C analogue, which has been stabilized against the serum nucleases that are present in the plasma of primates. Poly I:C(12)U, another analogue, is less toxic but also less stable in vivo than poly I:C, and TLR3 is essential for its recognition. To study the effects of these compounds on the induction of protein-specific immune responses in an animal model relevant to humans, rhesus macaques were immunized subcutaneously (s.c.) with keyhole limpet hemocyanin (KLH) or human papillomavirus (HPV)16 capsomeres with or without dsRNA or a control adjuvant, the TLR9 ligand CpG-C. All dsRNA compounds served as adjuvants for KLH-specific cellular immune responses, with the highest proliferative responses being observed with 2 mg/animal poly ICLC (p = 0.002) or 6 mg/animal poly I:C(12)U (p = 0.001) when compared with immunization with KLH alone. Notably, poly ICLC -- but not CpG-C given at the same dose -- also helped to induce HPV16-specific Th1 immune responses while both adjuvants supported the induction of strong anti-HPV16 L1 antibody responses as determined by ELISA and neutralization assay. In contrast, control animals injected with HPV16 capsomeres alone did not develop substantial HPV16-specific immune responses. Injection of dsRNA led to increased numbers of cells producing the T cell-activating chemokines CXCL9 and CXCL10 as detected by in situ hybridization in draining lymph nodes 18 hours after injections, and to increased serum levels of CXCL10 (p = 0.01). This was paralleled by the reduced production of the homeostatic T cell-attracting chemokine CCL21. Thus, synthetic dsRNAs induce an innate chemokine response and act as adjuvants for virus-specific Th1 and humoral immune responses in nonhuman primates.

Molecular analysis in true hermaphroditism: demonstration of low-level hidden mosaicism for Y-derived sequences in 46,XX cases.

True hermaphroditism (TH) is an unusual form of sex reversal, characterized by the development of testicular and ovarian tissue in the same subject. Approximately 60% of the patients have a 46,XX karyotype, 33% are mosaics with a second cell line containing a Y chromosome, while the remaining 7% are 46,XY. Molecular analyses have demonstrated that SRY is present in only 10% of TH with a 46,XX karyotype; therefore, in the remaining 90%, mutations at unknown X-linked or autosomal sex determining loci have been proposed as factors responsible for testicular development. True hermaphroditism presents considerable genetic heterogeneity with several molecular anomalies leading to the dual gonadal development as SRY point mutations or SRY hidden gonadal mosaicism. In order to identify genetic defects associated with subjects with the disease, we performed molecular analyses of the SRY gene in DNA from blood leukocytes and gonadal tissue in 12 true hermaphrodites with different karyotypes. Our results using PCR and FISH analyses reveal the presence of hidden mosaicism for SRY or other Y sequences in some patients with XX true hermaphroditism and confirms that mosaicism for SRY limited to the gonads is an alternative mechanism for testicular development in 46,XX true hermaphrodites.

Patrones de comorbilidad en pacientes psiquiatricos ambulatorios / Comorbidity patterns in psychiatric outpatients

En el presente estudio se determinaron patrones de comorbilidad en pacientes psiquiátricos ambulatorios en la Unidad de Salud Mental del Hospital San Juan de Dios (Bogotá). Se revisaron 3.028 historias clínicas de los pacientes atendidos en el servicio de consulta externa de Psiquiatría durante 1997, de estas se analizaron 1.127. Las variables que se tuvieron en cuenta fueron: edad, género y diagnóstico psiquiátrico en el eje I, II y III. Dada la naturaleza categórica de la mayor parte de las mediciones, se analizaron los datos mediante métodos multivariados de correspondencia múltiple. Los resultados mostraron la conformación de cuatro ejes: eje I mujeres, deprimidas, ansiosas, con comorbilidad cardiovascular y síndrome mental orgánico (SMO) y por otro lado hombres, psicosis, trastorno de personalidad o retardo mental. Eje II hombres con comorbilidad neurológica, SMO y trauma y por otra parte mujeres con comorbilidad endocrinológica y trastornos afectivos. En el eje III hombres con abuso de sustancias, trastornos de personalidad y diagnósticos relacionados con trauma o enfermedad cardiovascular. Por otro lado mujeres con patología endocrinológica, neurológica y retardo mental, sin diagnóstico en eje I. El eje IV recoge pacientes ansiosos, con trastornos de personalidad y alteraciones gastrointestinales o endocrinológica y por otra parte pacientes con trastorno bipolar o SMO y trauma

Acido Folico y depresion / Folic acid and major depressive disorder

La asociación entre deficiencia de ácido fólico y trastornos psiquiátricos ha sido estudiada y debatida desde el descubrimiento de esta vitamina en la década del 40. La relevancia clínica de esta deficiencia continúa siendo motivo de permanente investigación y ha sido inferida a partir de observaciones clínicas y de un mejor entendimiento del rol del folato en las vías metabólicas cerebrales. Este artículo hace una breve descripción de las características del acido fólico y su metabolismo, especialmente a nivel cerebral, así como una revisión de la literatura médica sobre la relación del ácido fólico y varios trastornos psiquiátricos, haciendo énfasis en el trastorno depresivo mayor