Iron deficiency in astrocytes alters cellular status and impacts on oligodendrocyte differentiation.

Iron deficiency (ID) has been shown to affect central nervous system (CNS) development and induce hypomyelination. Previous work from our laboratory in a gestational ID model showed that both oligodendrocyte (OLG) and astrocyte (AST) maturation was impaired. To explore the contribution of AST iron to the myelination process, we generated an in vitro ID model by silencing divalent metal transporter 1 (DMT1) in AST (siDMT1 AST) or treating AST with Fe3+ chelator deferoxamine (DFX; DFX AST). siDMT1 AST showed no changes in proliferation but remained immature. Co-cultures of oligodendrocyte precursors cells (OPC) with siDMT1 AST and OPC cultures incubated with siDMT1 AST-conditioned media (ACM) rendered a reduction in OPC maturation. These findings correlated with a decrease in the expression of AST-secreted factors IGF-1, NRG-1, and LIF, known to promote OPC differentiation. siDMT1 AST also displayed increased mitochondrial number and reduced mitochondrial size as compared to control cells. DFX AST also remained immature and DFX AST-conditioned media also hampered OPC maturation in culture, in keeping with a decrease in the expression of AST-secreted growth factors IGF-1, NRG-1, LIF, and CNTF. DFX AST mitochondrial morphology and number showed results similar to those observed in siDMT1 AST. In sum, our results show that ID, induced through two different methods, impacts AST maturation and mitochondrial functioning, which in turn hampers OPC differentiation.

Seeking the reuse of effluents and sludge from conventional wastewater treatment plants: Analysis of the presence of intestinal protozoa and nematode eggs.

Some of the microorganisms present in urban wastewater, which include intestinal protozoa and nematodes, can be pathogenic. Their (oo)cyst and egg transmissible stages are very resistant to environmental stresses and disinfectants and they are therefore difficult to remove. Thus, they can constitute a health risk if water or sludge obtained in the purification of wastewater is reused for agricultural purposes. In this context, the presence of intestinal protozoa and nematodes were studied in influents, effluents and sludge from five wastewater treatment plants (WWTPs) in the north of Spain by optical microscopy and PCR techniques. The removal efficiency of different wastewater treatments was also compared. The presence of protozoa has increased among the population discharging waste to WWTPs in recent years. Cryptosporidium spp., Giardia duodenalis, Entamoeba spp. and nematodes were detected in all of the WWTPs. Indeed, this is the first report of Entamoeba histolytica and Entamoeba moshkovskii in Spanish WWTPs. The water treatments studied showed different removal efficiencies for each species of intestinal protozoa, with the aerated lagoons providing the best results. (Oo)cysts were also detected in sludge even after aerobic digestion and dehydration. To avoid risks, (oo)cyst viability should be analysed whenever the sludge is to be used as a fertilizer. This study reinforces the necessity of establishing legal limits on the presence of protozoa in WWTP effluents and sludges, especially if reuse is planned. Further studies are necessary for a better understanding of the presence and behaviour of intestinal parasites.

[Factorial structure of the dysexecutive questionnaire in a sample of the Spanish population with acquired brain injury and memory deficit complaints]. / Estructura factorial del cuestionario disejecutivo en una muestra de población española con daño cerebral adquirido y quejas de déficit de memoria.

INTRODUCTION AND AIMS: The dysexecutive questionnaire (DEX) has been used to study executive deficits in both clinical samples (both psychiatric and neurological) and non-clinical samples, although agreement on its factorial structure is lacking. The aim of this research is to study that structure in the self-administered version in a sample of the Spanish population with acquired brain injury and memory loss complaints, and to compare that solution with those obtained in other studies. PATIENTS AND METHODS: The questionnaire was administered to 119 subjects with acquired brain injury with different aetiologies (traumatic, vascular, tumours, multiple sclerosis, toxic-metabolic and others). RESULTS: The Cronbach's alpha coefficient was 0.88. All the items showed adequate discriminatory power, except item 15. No relation was found between the total score on the DEX and the age, gender and time elapsed since the injury. A negative correlation between total score and level of schooling was confirmed. The total score does not follow a normal distribution. The five-factor solution accounts for a higher percentage of the total variance than those of two, three and four factors (63.76%). CONCLUSIONS: The Spanish version of the DEX is an instrument that is valid for evaluating general dysexecutive symptoms in subjects with acquired brain injury. The five-factor factorial structure (planning, cognitive control, inhibition, social awareness and impulse control) offers a greater wealth of information because it measures more aspects of the dys-executive pathology and therefore appears to be more useful in the clinical setting. It is advisable to use the questionnaire in an early stage of evaluation or screening and to use it as a complement to the proxy-reported version.

Prevalence of intestinal parasites in HIV-positive patients on the island of Bioko, Equatorial Guinea: its relation to sanitary conditions and socioeconomic factors.

The prevalence of intestinal parasitic diseases and their associated factors has been investigated in HIV populations from the Island of Bioko, Equatorial Guinea. The feces of 310 participants from the island of Bioko (260 HIV-positive and 50 HIV-negative) were analyzed by microscopic observation. Immunochromatography was also used to diagnose Giardia, Entamoeba histolytica and Cryptosporidium spp. In addition, patients were asked for sociodemographic, economic and academic status, and CD4+ T cell counts were recorded. For HIV-positive patients, the prevalence of infection by intestinal parasites was 81.5% (212/260), 83.8% (218/260) by pathogenic helminths and 55.4% (168/260) by pathogenic protozoa (E. histolytica/dispar and Giardia duodenalis). Gender association was found between the infection by Ascaris and Schistosoma, a higher proportion being found in women; and between Entamoeba and the place of residence, a higher proportion being observed in the urban belt. Strongyloides stercoralis and Chilomastix mesnili appeared only in the people of this group, all the cases of Chilomastix being in females. For HIV-negative participants, the prevalence of infection by intestinal parasites was 74.0% (37/50), 90.0% (45/50) by pathogenic helminths and 66.0% (43/50) by pathogenic protozoa. Gender, educational level and low hygiene were associated with intestinal parasitic infection. When comparing the two groups (HIV-positive and HIV-negative), statistical association between HIV co-infection and infection by Giardia and Entamoeba was found. Diarrhea was also associated with intestinal parasitic infection in the HIV-positive group. Not only do our findings reflect high rates of intestinal parasitic infections in HIV-positive people, but also in the HIV-negative group, suggesting a closer relationship between sanitary status and living conditions than with immune status, and thus they highlight the need to carry out health education policies in the population. In addition, Schistosoma intercalatum/guineensis has been detected for the first time as an autochthonous parasite on the island.

Evaluation of in vitro efficacy of combined riboflavin and ultraviolet a for Acanthamoeba isolates.

PURPOSE: To evaluate in vitro the amoebicidal effects of riboflavin and ultraviolet A (UVA) collagen cross-linking. DESIGN: Experimental study, laboratory investigation. METHODS: Two different strains of Acanthamoeba species were tested identically. Four treatment groups were considered: group 1 consisted of 0.1% riboflavin and 30-minute UVA irradiation; group 2 consisted of 0.1% riboflavin and 60-minute UVA irradiation; group 3 consisted of no riboflavin and no UVA exposure; group 4 consisted of 0.1% riboflavin and no UVA exposure. The application of UVA was performed under the parameters used for in vivo corneal collagen cross-linking. RESULTS: In all cases, cysts and trophozoites were detected 24 hours after treatment at a radial distance from the center of the seeding point more than 5 mm, indicating that the amoebae were viable. All treated and untreated groups of amoebae from the 2 strains exhibited growth (radii of 14 to 15 mm in groups 1, 3, and 4; radius of 12 mm in group 2). The final morphologic features of the 2 strains of trophozoites that received treatment were similar to those of the initial seeding group and the untreated control group. CONCLUSIONS: The results obtained in our study show that a single dose (30 or 60 minutes) of cross-linking cannot achieve eradication in the 2 different Acanthamoeba strains examined. However, in vitro results do not always indicate in vivo efficacy, so future studies should test the validity of this treatment for Acanthamoeba keratitis.

Identification and molecular characterization of Cryptosporidium and Giardia in children and cattle populations from the province of Álava, North of Spain.

The prevalence of and factors associated with the protozoan enteropathogens Cryptosporidium and Giardia have been investigated in selected children and cattle populations from the province of Álava (Northern Spain). The presence of these organisms was detected in fecal samples using commercially available coproantigen-ELISA (CpAg-ELISA) and immunochromatographic (ICT) assays. A total of 327 caregivers of children participants were asked to answer questions on risk factors potentially associated to the prevalence of Cryptosporidium and Giardia, including water-use practices, water sports and contact with domestic or pet animals. Molecular analyses were conducted using a nested-PCR technique to amplify the small-subunit (SSU) rRNA gene of Cryptosporidium and the triosephosphate isomerase (tpi) gene of Giardia. Cryptosporidium oocysts and Giardia cysts were found in 3 and 16 samples using the CpAg-ELISA, and in 5 and 9 samples using the ICT test, respectively. Cryptosporidium and Giardia were also found in 7 and 17 samples by CpAg-ELISA, and 4 and 14 samples by ICT, respectively, of 227 cattle fecal samples. The overall Cryptosporidium and Giardia infection prevalences, based on a Bayesian approach accounting for the imperfect sensitivities and specificities of both diagnostic tests, were estimated to 1.0% (95% BCI: 0.2%-2.8%) and 3.1% (1.5%-5.3%) in children and 3.0% (0.5%-9.2%) and 1.4% (0.0%-6.4%) in cattle, respectively. In humans, a single Cryptosporidium isolate was characterized as C. hominis. Of seven Giardia isolates, four were identified as assemblage B, two as assemblage A-II and one was a mixed assemblage B+A-II infection. No Cryptosporidium or Giardia isolates could be obtained from cattle samples. Although limited, these results seem to suggest that cattle are unlikely to be an important reservoir of zoonotic Cryptosporidium and/or Giardia in the province of Álava.