Multiomics plasma effects of switching from triple antiretroviral regimens to dolutegravir plus lamivudine.

INTRODUCTION: The DOLAM trial revealed that switching from triple antiretroviral therapy (three-drug regimen; 3DR) to dolutegravir plus lamivudine (two-drug regimen; 2DR) was virologically non-inferior to continuing 3DR after 48 weeks of follow-up. Weight increased with 2DR relative to 3DR but it did not impact on metabolic parameters. METHODS: Multiomics plasma profile was performed to gain further insight into whether this therapy switch might affect specific biological pathways. DOLAM (EudraCT 201500027435) is a Phase 4, randomized, open-label, non-inferiority trial in which virologically suppressed persons with HIV treated with 3DR were assigned (1:1) to switch to 2DR or to continue 3DR for 48 weeks. Untargeted proteomics, metabolomics and lipidomics analyses were performed at baseline and at 48 weeks. Univariate and multivariate analyses were performed to identify changes in key molecules between both therapy arms. RESULTS: Switching from 3DR to 2DR showed a multiomic impact on circulating plasma concentration of N-acetylmuramoyl-L-alanine amidase (Q96PD5), insulin-like growth factor-binding protein 3 (A6XND0), alanine and triglyceride (TG) (48:0). Correlation analyses identified an association among the up-regulation of these four molecules in persons treated with 2DR. CONCLUSIONS: Untargeted multiomics profiling studies identified molecular changes potentially associated with inflammation immune pathways, and with lipid and glucose metabolism. Although these changes could be associated with potential metabolic or cardiovascular consequences, their clinical significance remains uncertain. Further work is needed to confirm these findings and to assess their long-term clinical consequences.

Extending the Scope of the C-Functionalization of Cyclam via Copper(I)-Catalyzed Alkyne-Azide Cycloaddition to Bifunctional Chelators of Interest.

Cyclam, known for its potent chelation properties, is explored for diverse applications through selective N-functionalization, offering versatile ligands for catalysis, medical research, and materials science. The challenges arising from N-alkylation, which could decrease the coordination properties, are addressed by introducing a robust C-functionalization method. The facile two-step synthesis proposed here involves the click chemistry-based C-functionalization of a hydroxyethyl cyclam derivative using Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC). Boc-protecting groups prevent undesired copper coordination, resulting in compounds with a wide range of functionalities. The optimized synthesis conditions enable C-functional cyclams to be obtained easily and advantageously, with high application potential in the previously cited fields. The methodology has been extended to trehalose-based Siamese twin amphiphiles, enabling efficient gene delivery applications.

Prevalence and genetic diversity of azole-resistant Malassezia pachydermatis isolates from canine otitis and dermatitis: A 2-year study.

Despite previous reports on the emergence of Malassezia pachydermatis strains with decreased susceptibility to azoles, there is limited information on the actual prevalence and genetic diversity of azole-resistant isolates of this yeast species. We assessed the prevalence of azole resistance in M. pachydermatis isolates from cases of dog otitis or skin disease attended in a veterinary teaching hospital during a 2-year period and analyzed the ERG11 (encoding a lanosterol 14-α demethylase, the primary target of azoles) and whole genome sequence diversity of a group of isolates that displayed reduced azole susceptibility. Susceptibility testing of 89 M. pachydermatis isolates from 54 clinical episodes (1-6 isolates/episode) revealed low minimum inhibitory concentrations (MICs) to most azoles and other antifungals, but 11 isolates from six different episodes (i.e., 12.4% of isolates and 11.1% of episodes) had decreased susceptibility to multiple azoles (fluconazole, itraconazole, ketoconazole, posaconazole, ravuconazole, and/or voriconazole). ERG11 sequencing of these 11 azole-resistant isolates identified eight DNA sequence profiles, most of which contained amino acid substitutions also found in some azole-susceptible isolates. Analysis of whole genome sequencing (WGS) results revealed that the azole-resistant isolates from the same episode of otitis, or even different episodes affecting the same animal, were more genetically related to each other than to isolates from other dogs. In conclusion, our results confirmed the remarkable ERG11 sequence variability in M. pachydermatis isolates of animal origin observed in previous studies and demonstrated the value of WGS for disentangling the epidemiology of this yeast species.

We analyzed the prevalence and diversity of azole-resistant Malassezia pachydermatis isolates in a veterinary hospital. A low prevalence of multi-azole resistance (c.10% of isolates and cases) was found. Whole genome and ERG11 sequencing of resistant isolates revealed remarkable genetic diversity.

Analysis of the culturable gut yeast microbiota of dogs with digestive disorders.

Despite the increasing interest in studying the gut mycobiota of dogs, the association between fungal colonization and the development of digestive disorders in this species remains largely understudied. On the other hand, the high prevalence of antifungal-resistant yeasts detected in previous studies in samples from animals represents a major threat to public health. We analyzed the presence of culturable yeasts in 112 rectal swab samples obtained from dogs with digestive disorders attended in a veterinary teaching hospital. Our results revealed that Malassezia pachydermatis was frequently isolated from the studied dog population (33.9% of samples), and that the isolation of this yeast was significantly associated to the age of animals, but not to their sex, disease group, or the presence of vomits and/or diarrhea. In contrast, other yeast species were less prevalent (17.9% of samples in total), and their isolation was not significantly associated to any variable included in the analysis. Additionally, we observed that 97.5% of the studied M. pachydermatis isolates (n = 158, 1-6 per positive episode) displayed a minimum inhibitory concentration (MIC) value >4 µg/ml to nystatin, 31.6% had a MIC ≥32 µg/ml to fluconazole, and 27.2% had a MIC >4 µg/ml to amphotericin B. The antifungal susceptibility profiles of non-Malassezia (n = 43, 1-7 per episode) were more variable and included elevated MIC values for some antifungal-species combinations. These results confirm that the intestine of dogs is a reservoir of opportunistic pathogenic yeasts and suggest that the prevalence of M. pachydermatis colonization depends more on the age of animals than on any specific digestive disorder.

Trehalose-polyamine/DNA nanocomplexes: impact of vector architecture on cell and organ transfection selectivity.

A novel family of precision-engineered gene vectors with well-defined structures built on trehalose and trehalose-based macrocycles (cyclotrehalans) comprising linear or cyclic polyamine heads have been synthesized through procedures that exploit click chemistry reactions. The strategy was conceived to enable systematic structural variations and, at the same time, ensuring that enantiomerically pure vectors are obtained. Notably, changes in the molecular architecture translated into topological differences at the nanoscale upon co-assembly with plasmid DNA, especially regarding the presence of regions with short- or long-range internal order as observed by TEM. In vitro and in vivo experiments further evidenced a significant impact on cell and organ transfection selectivity. Altogether, the results highlight the potential of trehalose-polyamine/pDNA nanocomplex monoformulations to achieve targeting transfection without the need for any additional cell- or organ-sorting component.

Active surveillance of antimicrobial resistance in companion animals: A pilot study in a Spanish Veterinary Teaching Hospital.

The role of small animal veterinary hospitals in the onset and dissemination of antimicrobial-resistant organisms (AMROs) is still not clear, and the implementation of an internal surveillance systems is a cost-effective tool to better understand their impact. The aim of this study was to describe a pilot program of active surveillance in a Spanish Veterinary Teaching Hospital, developed to estimate the detection frequency of AMROs in the commensal flora of patients and in the environment. Surveillance was focused on Methicillin-resistant Staphylococci (MRS), third generation cephalosporins resistant gram-negative bacteria (3GCR-GNB), and carbapenems-resistant gram-negative bacteria (CR-GNB). Oral and perirectal swabs were collected in the same dogs and cats hospitalized > 48 h, at their admission and before their discharge. Out of 50 patients sampled, 24% (12/50) were carriers at admission of at least one of the three investigated AMROs. Twenty-eight percent of patients (14/50) acquired at least one AMRO during the hospital stay. MRS detection frequency at admission was 12% (6/50), while acquisition was 6% (3/50). 3GCR-GNB detection frequency was 14% at admission (7/50) and acquisition 22% (11/50), while CR-GNB detection frequency was 2% at admission (1/50) and acquisition 2% (1/50). Environmental surveillance (98 samples) showed a total detection frequency of 22.4% for MRS (22/98), 2% for 3GCR-GNB and CR-GNB (2/98). Clinical staff' shoe soles showed high detection frequency for MRS (50%). 3GCR Escherichia coli was the most isolated species in patients (n = 17). The results show how active surveillance can be used as a tool to assess the impact of AMROs in veterinary hospitals to subsequently build up tailored control plans based on specific issues.

Crecimiento fúngico en medios de cultivo que simulan un ambiente extremo / Fungal growth in culture media simulating an extreme environment

Antecedentes. Hay un creciente interés por el estudio de los microorganismos que habitan ambientes extremos por razones que van desde incrementar el conocimiento sobre el origen de la vida hasta la búsqueda de nuevas aplicaciones biotecnológicas. Objetivos. En el presente trabajo se aborda el estudio de la tolerancia de hongos aislados del Arroyo de Aguas Agrias (AAS; Tharsis, Huelva, España), un ambiente ácido y rico en metales, frente a medios de cultivo preparados con agua procedente de este ecosistema extremo (medio AASW). También se investigó la posibilidad de crecimiento en estas condiciones de cepas de colección de hongos y levaduras. Métodos. Para los hongos miceliares se calculó un índice de tolerancia, definido como el cociente entre el diámetro de crecimiento de las colonias sobre AASW y el que se produce en un medio control. Para las levaduras se determinó la concentración mínima inhibitoria de AASW. Resultados. En general, los hongos aislados del AAS manifestaron diferencias en su capacidad para germinar y crecer sobre el medio AASW. Las cepas de colección del género Aspergillus fueron capaces de crecer sobre el medio AASW, pero mostraron diferencias en su tolerancia al mismo en comparación con los aislamientos ambientales. Conclusiones. Los hongos extremotolerantes pueden manifestar diferencias en su tolerancia a medios de cultivo que simulan las condiciones de su hábitat natural. Los resultados de nuestro trabajo sugieren que la capacidad de los hongos para crecer en ambientes ácidos, ricos en metales, puede ser más común de lo que pudiera pensarse, y pone de manifiesto la importancia de determinar los factores específicos que son responsables de la tolerancia a esos ambientes extremos(AU)

Background. There is an increasing interest in the study of microorganisms that inhabit extreme environments for reasons that vary from gaining insight into the origin of life to the searching of new biotechnological applications. Aims. In this work, we studied the tolerance of fungi isolated from the Aguas Agrias Stream (AAS; Tharsis, Huelva, Spain), an acidic metal-rich environment, to a culture medium prepared with water from this extreme ecosystem (AASW medium). The ability of some culture collection strains of moulds and yeasts to grow on AASW medium was also assessed. Methods. For moulds, a tolerance index was calculated by dividing the growth diameter of colonies on AASW medium by the diameter in the control medium, and their germinative potential was recorded. For yeasts and yeast-like fungi, the minimum inhibitory concentration of AASW was determined. Results. In general, the fungi isolated from the AAS showed differences in their ability to germinate and grow on AASW medium. Collection strains of the genus Aspergillus could grow on AASW medium, but showed some differences in tolerance when compared to environmental isolates. Conclusions. Extremotolerant fungi can manifest differences in their tolerance to culture media that simulate the conditions of their natural habitat. The results of this work suggest that the ability of fungi to grow in acidic, metal-rich environments might be more widespread than previously thought, and highlight the importance of determining the factors that are responsible for tolerance to these extreme environments(AU)

Primary human immunodeficiency virus type 1 infection: clinical, virological and immunological characteristics of 75 patients (1997-2003)

Objetivos. Describir las características epidemiológicas, clínicas y evolutivas de una cohorte de pacientes con una infección aguda por el virus de la inmunodeficiencia humana (VIH) en el área de Barcelona. Métodos. Estudio prospectivo de pacientes diagnosticados de infección aguda por el VIH en un hospital terciario de Barcelona durante el período 1997-2003. Análisis descriptivo de las características epidemiológicas y clínicas e influencia del tratamiento antirretroviral (TARV) en la evolución. Resultados. Se diagnosticaron 75 pacientes, lo que representó el 2,9% del total de pacientes diagnosticados de infección por el VIH en el mismo período de tiempo. El 81% eran varones y la mediana de edad fue de 30 años (rango intercuartil [RIC], 26-38). Las vías de contagio fueron las relaciones homosexuales (72%), seguida de las heterosexuales (17%) y del uso de drogas intravenosas (11%). El 77% de los pacientes presentó síntomas, siendo los más frecuentes: fiebre (98%), astenia (86%), artromialgias (65%), linfoadenopatías (55%), sudoración nocturna (48%) y exantema (45%). El 65% comenzó TARV, disminuyendo el número de pacientes tratados del 79% en el período 1997-2000 al 49% en el período 2001-2003 (p < 0,01). Tras una mediana de seguimiento de 37 meses (RIC, 26-66), un paciente falleció y 8 casos se perdieron de seguimiento. Los pacientes que no recibieron TARV presentaron una mayor probabilidad de presentar deterioro inmunológico o clínico durante el seguimiento en comparación con el grupo que recibió TARV (42,3% frente a 12,3%; p < 0,001). La dislipemia y la lipodistrofia se diagnosticaron en el 58 y 37% de los pacientes tratados, respectivamente. Conclusiones. La infección aguda por VIH se diagnosticó con más frecuencia en los varones homosexuales, siendo sus características clínicas similares a las descritas previamente. El TARV instaurado en esta fase de la infección por VIH fue eficaz pero se asoció a una frecuencia elevada de efectos adversos (AU)

Objectives. To describe the epidemiological and clinical characteristics and the evolution of a cohort of patients with primary HIV-1 infection from the Barcelona area. Methods. Prospective cohort study of HIV-infected patients diagnosed with primary HIV infection in a tertiary hospital in Barcelona (Spain) from 1997 through 2003. Descriptive analysis of epidemiological and clinical characteristics and effect of highly active antiretroviral treatment (HAART) on outcome. Results. A total of 75 patients were diagnosed, accounting for 2.9% of the total of newly diagnosed HIV patients during the same time period. Eighty-one percent of the patients were males and the median age was 30 years (IQR 26-38). The most frequent transmission route was homosexual (72%), followed by heterosexual (17%) and intravenous drug abuse (11%). Seventy-seven percent of patients presented symptoms, the most frequent being fever (98%), asthenia (86%), arthralgia-myalgia (65%), lymphadenopathy (55%), night sweats (48%) and rash. Sixty-five percent started HAART, although the proportion of patients that received HAART decreased from 79% during the period 1997-2000 to 49% during the period 2001-2003 (p < 0.01). After a median follow-up of 37 months (IQR 26-66), one patient died and eight cases were lost to follow-up. The patients who did not receive HAART had a higher probability of immunological or clinical deterioration during the follow-up when compared to the group that received HAART (42.3% versus 12.3%; p < 0.001). In treated patients, dyslipidemia and lipodystrophy were diagnosed in 58% and 37% of cases, respectively. Conclusions. Primary HIV-1 infection was diagnosed more frequently in homosexual males, and its clinical characteristics were similar to those observed in previous studies. HAART given during primary HIV infection was effective, but was associated with a high percentage of adverse effects (AU)

Avances en el diagnóstico y tratamiento de la infección por el VHC en pacientes con y sin infección por el VIH / Advances in the diagnosis and treatment of hepatitis C virus infection in patients with and without HIV infection

La infección crónica por el virus de la hepatitis C (VHC) representa un grave problema sanitario que afecta al 1-3% de la población mundial. Se transmite por vía sexual, vertical y de forma primordial tras exposición a sangre por vía percutánea. Dado que comparte vías de contagio similares a las del virus de la inmunodeficiencia humana (VIH), la coinfección VIH-VHC es muy frecuente y la hepatopatía crónica, así como las complicaciones asociadas a su curso clínico, son una importante causa de morbimortalidad en esta población. El pilar del tratamiento para el VHC ha sido el interferón al que posteriormente se le ha añadido ribavirina. En la actualidad la combinación de ribavirina y una nueva formulación pegilada del interferón constituye la terapia estándar con la que se consiguen tasas de respuesta viral sostenida del 40-80% (AU)

The chronic infection by the hepatits C virus represents a serious sanitary problem affecting 1-3% of the world-wide population. It is transmitted by sexual route, vertical route and mainly after blood exposure by percutanea route. While HIV shares similar routes of transmission, the co-infection HCV-HIV is very frequent and the chronic hepatopathy and complications associated with its clinical course are an important cause of morbi-mortality in this population. The gold standard of the treatment for the HCV, has been the interferon and later the combination therapy of interferon plus ribavirine. Currently, the combination of ribavirine and a new pegilated formulation of the interferon has become the standard in the treatment reaching rates of sustained viral response around 40-80% (AU)

Trasplante renal en pacientes infectados por el VIH / Renal transplantation in patients with HIV infection

La prevalencia de la infección por el virus de la inmunodeficiencia humana (VIH) en pacientes en terapia renal sustitutiva (TRS) es muy variable, pero de forma global se estima que en Europa es del 1% y en Estados Unidos del 1,5%. La supervivencia de estos pacientes en TRS también ha mejorado notablemente con la introducción de tratamiento antirretroviral de gran actividad (TARGA). La experiencia acumulada en la era del TARGA en trasplante renal en pacientes infectados por el VIH en Estados Unidos indica que la supervivencia a los 3 años es similar a la de los pacientes sin infección por el VIH, con un buen control virológico e inmunológico de la infección por el VIH bajo TARGA y sin presentar mayor número de infecciones oportunistas y/o tumores. Los criterios de selección de pacientes infectados por el VIH que han utilizado los diferentes grupos de trasplante han sido: ausencia de manifestaciones oportunistas previas, tener una cifra de linfocitos CD41 superior a 200 cél./μl y una carga viral del VIH suprimible con TARGA. En España, donde la mayoría de pacientes eran antiguos drogadictos, para el trasplante hepático se exigiría además una abstinencia de heroína y cocaína de 2 años de duración, y el paciente podía estar en el programa de metadona. Los principales problemas detectados en el período postrasplante son las interacciones farmacocinéticas y farmacodinámicas entre los antirretrovirales y los inmunosupresores, el manejo de la coinfección por el virus de la hepatitis C y la elevada tasa de rechazo. En España se han realizado hasta la fecha siete trasplantes renales, con buena evolución del paciente y del injerto y sin progresión de la infección por el VIH (AU)

The prevalence of human immunodeficience virus (HIV) infection among patients under renal replacement therapy varies, with estimates of 1% for Europe and 1.5% for the United States. Survival in HIV infected individuals receiving renal replacement therapy has improved since the introduction of high activity antiretroviral therapy (HAART). Current experience in renal transplantation in HIV-infected patients in the United States indicates that the three-year survival rate is similar to that of HIV-negative transplant recipients, with virological and immunological control of the infection by HAART and no increase in the number of opportunistic infections or tumors. The criteria for selecting renal transplantation candidates in this population are the following: no aids-defining events, CD4 cells > 200 cells/μl and undetectable viral load under HAART. In Spain, where most of these patients are former drug abusers, a two-year period of abstinence from cocaine and heroine abuse is also required, although patients can be participating in the methadone program. The main problems in the post-transplantation period have been interactions between HAART and immunosuppressive drugs, management of hepatitis C virus (HCV) coinfection and the high rate of acute rejection. To date, seven such renal transplantations have been performed in Spain, with favorable patient and graft survival and no progression to aids (AU)