Primary Sclerosing Cholangitis: Gender Effects in Valencia's Low-Prevalence Region.

BACKGROUND AND AIMS: Recent studies point out to epidemiological changes in primary sclerosing cholangitis (PSC). Our aims were to determine in PSC patients followed in several centers in a Mediterranean geographic area: (i) changes in baseline features and (ii) effect of gender on clinical course. METHODS: Retrospective multicenter study of PSC patients treated in 8 hospitals in a Mediterranean area between 2000 and 2021. Charts were reviewed compiling demographic, clinical, radiological, and histological variables. RESULTS: Cohort of 112 PSC patients included, 42% women, 70% diagnosed after 2010. Women were increasingly diagnosed in recent cohorts. The median time from diagnosis to the combined endpoint liver transplantation (Lt) and/or death was 6.9 years. Asthenia at diagnosis (p = 0.009) was associated with lower transplant-free survival, while diagnosis before 2005 was associated with greater LT-free survival (p < 0.001). By Cox regression, LT-free survival was not influenced by age, sex, or cirrhosis at the time of diagnosis. Women were found to have less jaundice at diagnosis (2 vs 14%; p = 0.013), higher prevalence of ANA antibodies (43.9 vs 15.7%; p = 0.003), and lower GGT levels at diagnosis (GGT 123 vs 209U/L; p = 0.014) than men. CONCLUSION: In an area traditionally considered to have low prevalence, the prevalence of affected women surpasses expectations based on existing literature. There appear to be gender-related variations in the presentation of the condition, highlighting the need for confirmation through larger-scale studies.

Design, synthesis and biological evaluation of antiparasitic dinitroaniline-ether phospholipid hybrids.

A series of nine novel ether phospholipid-dinitroaniline hybrids were synthesized in an effort to deliver more potent antiparasitic agents with improved safety profile compared to miltefosine. The compounds were evaluated for their in vitro antiparasitic activity against L. infantum, L.donovani, L. amazonensis, L. major and L. tropica promastigotes, L. infantum and L. donovani intracellular amastigotes, Trypanosoma brucei brucei and against different developmental stages of Trypanosoma cruzi. The nature of the oligomethylene spacer between the dinitroaniline moiety and the phosphate group, the length of the side chain substituent on the dinitroaniline and the choline or homocholine head group were found to affect both the activity and toxicity of the hybrids. The early ADMET profile of the derivatives did not reveal major liabilities. Hybrid 3, bearing an 11-carbon oligomethylene spacer, a butyl side chain and a choline head group, was the most potent analogue of the series. It exhibited a broad spectrum antiparasitic profile against the promastigotes of New and Old World Leishmania spp., against intracellular amastigotes of two L. infantum strains and L. donovani, against T. brucei and against T. cruzi Y strain epimastigotes, intracellular amastigotes and trypomastigotes. The early toxicity studies revealed that hybrid 3 showed a safe toxicological profile while its cytotoxicity concentration (CC50) against THP-1 macrophages being >100 µM. Computational analysis of binding sites and docking indicated that the interaction of hybrid 3 with trypanosomatid α-tubulin may contribute to its mechanism of action. Furthermore, compound 3 was found to interfere with the cell cycle in T. cruzi epimastigotes, while ultrastructural studies using SEM and TEM in T. cruzi showed that compound 3 affects cellular processes that result in changes in the Golgi complex, the mitochondria and the parasite's plasma membrane. The snapshot pharmacokinetic studies showed low levels of 3 after 24 h following oral administration of 100 mg/Kg, while, its homocholine congener compound 9 presented a better pharmacokinetic profile.

Etoricoxib-induced fixed drug eruption: Report of seven cases.

BACKGROUND: Fixed drug eruption (FDE) is a characteristic form of intraepidermal CD8+ T cell-mediated drug reaction, with repeated appearance of isolated or multiple skin lesions in the same location after receiving the offending drug. Non-steroidal anti-inflammatory drugs (NSAID) are the most common cause. Selective inhibitors of inducible cyclooxygenase 2 (COX-2) provoke a lesser degree of allergic or idiosyncratic adverse reactions than conventional NSAID, but they can cause skin reactions of variable severity. OBJECTIVE: Etoricoxib has been related to a variety of unusual skin reactions, including several reports of FDE. METHODS: We perfomed epicutaneous test to diagnose patients with suspected etoricoxib fixe drug rash due to clinical features and reproducibility on at least two occasions. RESULTS: We present seven new cases of etoricoxib-induced fixed drug eruption, with a diagnosis based on clinical presentation. This diagnosis was confirmed by an etoricoxib-positive lesional patch test in six cases and by a positive low-dose oral challenge in the other one. Two patients showed negative patch tests with celecoxib (10% in pet.) on the residual lesions, and oral tolerance was confirmed in one. CONCLUSION: To our knowledge, this is the largest series on FDE induced by etoricoxib reported to date.

In-Vitro Activity of Silybin and Related Flavonolignans against Leishmania infantum and L. donovani.

Flavonolignans from the seeds of the milk thistle (Silybum marianum) have been extensively used in folk medicine for centuries. Confirmation of their properties as hepatoprotective, antioxidant and anticancer has been obtained using standardized extracts and purified flavonolignans. Information on their potential effect on Leishmania is very scarce. We have investigated the effect of silymarin, silybin and related flavonolignans on the multiplication of promastigotes in vitro and ex vivo on intracellular amastigotes of L. infantum (Li) and L. donovani (Ld), causative agents of human and canine visceral leishmaniasis (VL). In addition, the potential synergistic effect of the most active molecule and well-established antileishmanial drugs against promastigotes was explored. Dehydroisosilybin A elicited the highest inhibition against Ld and Li promastigotes with an approximate IC50 of 90.23 µM. This molecule showed a moderate synergism with amphotericin B (AmB) but not with SbIII or paromomycin, although it was ineffective against amastigotes. Antileishmanial activity on intracellular amastigotes of the two diastereoisomers of dehydrosilybin (10 µM) was comparable to that elicited by 0.1 µM AmB. Antiproliferative activity and safety of flavonolignans suggest the interest of exploring their potential value in combination therapy against VL.

The lithic industry of Sima del Elefante (Atapuerca, Burgos, Spain) in the context of Early and Middle Pleistocene technology in Europe.

This paper presents the lithic assemblages documented at Sima del Elefante (TE) and their importance in the context of the Early and Middle Pleistocene human occupation of Europe. We also study changes in human behaviour within the context of the palaeoenvironmental evolution of the Sierra de Atapuerca. This site has characteristics that are of great value for the study of human evolution. The lower levels of TE (Units TE7-TE14) are an essential reference for understanding the early stages of the colonization of Europe. The TE9c level has provided stone tools (Mode 1), faunal remains, and human fossils dated to 1.22 Ma (millions of years ago). Moreover, this is one of the few European sites with a stratigraphic sequence that includes remains of human occupations predating the Jaramillo subchron (Early Pleistocene) and from the Late Middle Pleistocene (Units TE18-TE19). Despite this, the presence of archaeologically sterile units (TE15-17) prevents us from establishing a continuous relationship between the Early and Middle Pleistocene human settlements and, consequently, between their technological and behavioural differences. We can, however compare the technological and palaeoeconomic strategies adopted by different species of hominins during two key phases of the occupation of Europe.

Safety and efficacy of treatment with pegylated interferon alpha-2a with ribavirin in chronic hepatitis C genotype 4.

The hepatitis C virus (HCV) genotype is an important predictive outcome parameter for pegylated interferon plus ribavirin therapy. Most published therapeutic trials to date have enrolled mainly patients with HCV genotypes 1, 2 and 3. Limited studies have focused on genotype 4 patients, who have had a poor representation in pivotal trials. Our aim was to evaluate the efficacy and safety of treatment with standard dose pegylated interferon alfa-2a in combination with weight-based ribavirin in patients with chronic hepatitis C genotype 4. In this prospective observational study, 198 patients with HCV-4 were included in this study from February 2004 to August 2005,188 patients who received at least 1 dose of drugs were included in the ITT analysis and they were treated with pegylated interferon alfa-2a and ribavirin for 48 weeks. Baseline and demographic characteristics, response to treatment at weeks 12, 48 and 72, and the nature and frequency of adverse effects were analyzed. Virological response at week 12 was achieved in 144 patients (76.6%). Virological response at the end of treatment was present in 110 patients (58.5%). At week 72, 99 patients presented SVR (52.7%). The reported adverse events were similar to those found in the literature for treatments of similar dose and duration. In conclusion, combined treatment with pegylated interferon alfa-2a and ribavirin was well tolerated and effective in chronic hepatitis C genotype 4, yielding response rates between those reported for genotype 1 and those of genotypes 2-3.

Experimental evidence of harmful effects of Crithidia mellificae and Lotmaria passim on honey bees.

The trypanosomatids Crithidia mellificae and Lotmaria passim are very prevalent in honey bee colonies and potentially contribute to colony losses that currently represent a serious threat to honey bees. However, potential pathogenicity of these trypanosomatids remains unclear and since studies of infection are scarce, there is little information about the virulence of their different morphotypes. Hence, we first cultured C. mellificae and L. passim (ATCC reference strains) in six different culture media to analyse their growth rates and to obtain potentially infective morphotypes. Both C. mellificae and L. passim grew in five of the media tested, with the exception of M199. These trypanosomatids multiplied fastest in BHI medium, in which they reached a stationary phase after around 96 h of growth. Honey bees inoculated with either Crithidia or Lotmaria died faster than control bees and their mortality was highest when they were inoculated with 96 h cultured L. passim. Histological and Electron Microscopy analyses revealed flagellated morphotypes of Crithidia and Lotmaria in the lumen of the ileum, and adherent non-flagellated L. passim morphotypes covering the epithelium, although no lesions were evident. These data indicate that parasitic forms of these trypanosomatids obtained from the early stationary growth phase infect honey bees. Therefore, efficient infection can be achieved to study their intra-host development and to assess the potential pathogenicity of these trypanosomatids.

[Hepatic steatosis in chronic hepatitis C: study of risk factors and relationship with the fibrosis stage]. / Esteatosis hepática en la hepatitis crónica por virus C: estudio de los factores de riesgo y relación con el estadio de fibrosis.

BACKGROUND AND OBJECTIVE: Liver steatosis (LS) is a frequent histological finding in chronic hepatitis C virus (HCV) infection with prognostic implications. The aim of the present prospective study was to analyse the risk factors of steatosis and its relationship with the fibrosis stage in patients with chronic HCV infection. MATERIAL AND METHOD: Eighty four consecutive HCV RNA positive patients, not treated previously, in whom a liver biopsy was performed, were enrolled. In each patient demographic, clinical, laboratory, viral, and histological variables were obtained at the time of biopsy. Bivariate and multivariate analysis, calculating the odds ratio (OR) and the 95% confidence interval (95%CI), were performed. RESULTS: LS was present in 69% of patients. Risk factors of steatosis were an increase of the body mass index (OR: 1.17; 95%CI: 1.01-1.35) and chronic alcohol consumption (OR: 3.58; 95%CI: 1.1-11.6) whereas those of fibrosis were chronic alcohol consumption (OR: 3.58; 95%CI: 1.1-11.6) and increase of the liver inflammatory activity (OR: 1.31; 95%CI: 1.13-1.53). LS was associated with genotype 3 virus infection, which was present in all patients with this infection who had severe steatosis in a significantly greater proportion than in patients with non-genotype 3 virus infection (41.7% vs 2.8%; P<.001). LS was more frequent in patients with advanced fibrosis stages than in patients with non-advanced fibrosis (78,9% vs 60,9%; P=.074). CONCLUSIONS: LS is a frequent finding in HCV chronic infection related to both host and viral factors. LS could be a worsening factor of hepatic injury.

Early antibody response and clinical outcome in experimental canine leishmaniasis.

Infected dogs are the main reservoir of zoonotic visceral leishmaniasis, a widespread parasitic disease caused by Leishmania infantum. Therefore, the control of canine infections is required to reduce the incidence of human cases. Disease outcome in dogs depends on the fine balance between parasite virulence and efficacy of the immune system. Thus, knowledge of early response could yield relevant information for diagnosis and follow-up. In our study, 20 Beagle dogs were intravenously infected with 108 amastigotes of a fresh isolate of L. infantum and monitored along 16 weeks post inoculation. Specific antibody response and clinical evolution of infected animals were highly variable. Immunofluorescence antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) were useful to assess infection status, although only ELISA with promastigote-coated plates and, particularly, western blotting (WB) allowed an early diagnosis. Prominent antigens were identified by mass peptide fingerprinting. Chaperonin HSP60, 32 and 30 KDa antigens were recognized by all dogs on week 10 post infection. This suggests that these antigens may be valuable for early diagnosis. Advanced infection showed, in addition, reactivity to HSP83 and HSP70. Disease outcome did not show a clear relationship with ELISA or IFAT titers. Correlation between the clinical status and the combined reactivity to some antigens sustains their use for diagnosis and follow-up.

Syrian Hamster as an Advanced Experimental Model for Visceral Leishmaniasis.

Animal models are needed along the development and evaluation of potential chemotherapeutic agents against leishmaniasis. Infections of Syrian hamsters with Leishmania species causing visceral leishmaniasis (VL) closely mimic the disease in the natural hosts, including target organs, lesions, and clinical course. Therefore, despite some shortcomings (e.g., genetic background, price, and scarcity of reagents), it is probably the best laboratory rodent model of VL. However, handling of hamsters can be technically challenging because of their particular anatomy. Here, we describe in detail four different routes to establish an experimental VL in the hamster model using Leishmania promastigotes and amastigotes. Each route requires various manipulations and has different benefits and drawbacks. Choice of the most suitable route should be made by the researcher in accordance with the specific plan and purpose of the study.

SAR Studies and Biological Characterization of a Chromen-4-one Derivative as an Anti-Trypanosoma brucei Agent.

Chemical modulation of the flavonol 2-(benzo[d][1,3]dioxol-5-yl)-chromen-4-one (1), a promising anti-Trypanosomatid agent previously identified, was evaluated through a phenotypic screening approach. Herein, we have performed structure-activity relationship studies around hit compound 1. The pivaloyl derivative (13) showed significant anti-T. brucei activity (EC50 = 1.1 µM) together with a selectivity index higher than 92. The early in vitro ADME-tox properties (cytotoxicity, mitochondrial toxicity, cytochrome P450 and hERG inhibition) were determined for compound 1 and its derivatives, and these led to the identification of some liabilities. The 1,3-benzodioxole moiety in the presented compounds confers better in vivo pharmacokinetic properties than those of classical flavonols. Further studies using different delivery systems could lead to an increase of compound blood levels.

Discovery of a benzothiophene-flavonol halting miltefosine and antimonial drug resistance in Leishmania parasites through the application of medicinal chemistry, screening and genomics.

Leishmaniasis, a major health problem worldwide, has a limited arsenal of drugs for its control. The appearance of resistance to first- and second-line anti-leishmanial drugs confirms the need to develop new and less toxic drugs that overcome spontaneous resistance. In the present study, we report the design and synthesis of a novel library of 38 flavonol-like compounds and their evaluation in a panel of assays encompassing parasite killing, pharmacokinetics, genomics and ADME-Toxicity resulting in the progression of a compound in the drug discovery value chain. Compound 19, 2-(benzo[b]thiophen-3-yl)-3-hydroxy-6-methoxy-4H-chromen-4-one, exhibited a broad-spectrum activity against Leishmania spp. (EC50 1.9 µM for Leishmania infantum, 3.4 µM for L. donovani, 6.7 µM for L. major), Trypanosoma cruzi (EC50 7.5 µM) and T. brucei (EC50 0.8 µM). Focusing on anti-Leishmania activity, compound 19 challenge in vitro did not select for resistance markers in L. donovani, while a Cos-Seq screening for dominant resistance genes identified a gene locus on chromosome 36 that became ineffective at concentrations beyond EC50. Thus, compound 19 is a promising scaffold to tackle drug resistance in Leishmania infection. In vivo pharmacokinetic studies indicated that compound 19 has a long half-life (intravenous (IV): 63.2 h; per os (PO): 46.9 h) with an acceptable ADME-Toxicity profile. When tested in Leishmania infected hamsters, no toxicity and limited efficacy were observed. Low solubility and degradation were investigated spectroscopically as possible causes for the sub-optimal pharmacokinetic properties. Compound 19 resulted a specific compound based on the screening against a protein set, following the intrinsic fluorescence changes.

Tratamiento con ribavirina de la infección crónica por el virus de la hepatitis E en pacientes trasplantados / Treatment with ribavirin of hepatitis E virus chronic infection in transplanted patients

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Papel de la homocisteína en la esteatosis hepática asociada al virus de la hepatitis C / Role of homocysteine in liver steatosis associated with virus C hepatitis

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Esteatosis hepática en la hepatitis crónica por virus C: estudio de los factores de riesgo y relación con el estadio de fibrosis / Hepatic steatosis in chronic hepatitis C: study of risk factors and relationship with the fibrosis stage

Fundamento y objetivo: La esteatosis hepática (EH) es un hallazgo histológico frecuente en la infección crónica por el virus de la hepatitis C (VHC) que puede influir en el pronóstico de la enfermedad. El objetivo del presente estudio prospectivo es valorar los factores asociados a su presencia y su relación con el estadio de fibrosis en pacientes con infección crónica por VHC. Material y método: Se incluyó a 84 pacientes consecutivos con ARN-VHC positivo, no tratados previamente y a los que se practicó una biopsia hepática. En todos los pacientes se recogieron en el momento de la biopsia variables demográficas, clínicas, analíticas, virales e histológicas. Se practicaron análisis bivariante y multivariante, calculando la odds ratio (OR) y el intervalo de confianza (IC) del 95%. Resultados: La EH estuvo presente en 69% de los pacientes. Los factores de riesgo de esteatosis fueron el aumento del índice de masa corporal (OR=1,17; IC del 95%, 1,01¿1,35) y el consumo crónico de alcohol (OR=5,73; IC del 95%, 1,12¿29,63) mientras los de fibrosis avanzada fueron el consumo crónico de alcohol (OR=3,58; IC del 95%, 1,1¿11,6) y el aumento de la actividad inflamatoria hepática (OR=1,31; IC del 95%, 1,13¿1,53). La EH se asoció al genotipo viral 3, ya que estuvo presente en todos los pacientes con este genotipo, los cuales presentaron esteatosis grave en una proporción significativamente superior a la presentada por el resto de los pacientes (el 41,7 frente al 2,8%; p<0,001). La frecuencia de esteatosis fue mayor en los pacientes con estadios avanzados de fibrosis que en el resto de pacientes (el 78,9 frente al 60,9%; p=0,074). Conclusiones: La EH es un hallazgo frecuente en la infección crónica por VHC que se relaciona tanto con factores del huésped como virales. La presencia de esteatosis puede ser un factor de agravamiento de la lesión hepática (AU)

Background and objective: Liver steatosis (LS) is a frequent histological finding in chronic hepatitis C virus (HCV) infection with prognostic implications. The aim of the present prospective study was to analyse the risk factors of steatosis and its relationship with the fibrosis stage in patients with chronic HCV infection. Material and method: Eighty four consecutive HCV RNA positive patients, not treated previously, in whom a liver biopsy was performed, were enrolled. In each patient demographic, clinical, laboratory, viral, and histological variables were obtained at the time of biopsy. Bivariate and multivariate analysis, calculating the odds ratio (OR) and the 95% confidence interval (95%CI), were performed. Results: LS was present in 69% of patients. Risk factors of steatosis were an increase of the body mass index (OR: 1.17; 95%CI: 1.01¿1.35) and chronic alcohol consumption (OR: 3.58; 95%CI: 1.1¿11.6) whereas those of fibrosis were chronic alcohol consumption (OR: 3.58; 95%CI: 1.1¿11.6) and increase of the liver inflammatory activity (OR: 1.31; 95%CI: 1.13¿1.53). LS was associated with genotype 3 virus infection, which was present in all patients with this infection who had severe steatosis in a significantly greater proportion than in patients with non-genotype 3 virus infection (41.7% vs 2.8%; P<.001). LS was more frequent in patients with advanced fibrosis stages than in patients with non-advanced fibrosis (78,9% vs 60,9%; P=.074). Conclusions: LS is a frequent finding in HCV chronic infection related to both host and viral factors. LS could be a worsening factor of hepatic injury (AU)