Pyriproxyfen Contamination in Daphnia magna: Identifying Early Warning Biomarkers.

Pyriproxyfen is an insecticide currently employed in numerous countries for the management of agricultural and indoor pests. Several studies indicate that this insecticide has been detected in multiple rivers, with concentrations reaching as high as 99.59 ng/L in the Júcar River in Spain. Therefore, the determination of some biochemical and genetic effects of this insecticide on aquatic organisms could serve as an early warning mechanism to identify potential disruptions in various biomarkers. Based on this, Daphnia magna organisms were exposed to pyriproxyfen sublethal concentrations for 21 days. Some biochemical parameters, including cholesterol, triglycerides, glucose, lactate, and LDH activity, were determined. Additionally, some genetic biomarkers associated with oxidative stress, heat shock proteins, lipid metabolism, hemoglobin, metallothioneins, and vitellogenin synthesis were evaluated in daphnids exposed to the insecticide for 21 days. LDH activity increased significantly in those daphnids exposed to the highest insecticide concentration (14.02 µg/L), while cholesterol levels decreased significantly. In contrast, glucose, total proteins, and triglycerides remained unaffected in D. magna exposed to pyriproxyfen. On the other hand, exposure to the insecticide led to notable alterations in gene expression among individuals. Specifically, genes associated with lipid metabolism and reproduction exhibited a significant reduction in gene expression. Fabd expression was decreased by approximately 20% in exposed daphnids, while vtg expression was suppressed as much as 80% when compared to control values. Furthermore, it was observed that the hgb1 and hgb2 genes, associated with hemoglobin synthesis, exhibited significant overexpression. Notably, the dysfunction observed in both hemoglobin genes was linked to an increase in pigmentation in Daphnia magna during the course of the experiment. These alterations in gene expression could serve as effective indicators of early contamination even at low pesticide concentrations.

Pediatric renal lithiasis in Spain: research, diagnostic and therapeutic challenges, and perspectives.

Incidence and prevalence of urolithiasis is apparently increasing worldwide, also among children and adolescents. Nevertheless, robust data have only been obtained in a few countries. In Spain, a voluntary Registry for Pediatric Renal Lithiasis has been active since 2015. Irregular participation limits its applicability, as well as its limitation to patients with a stone available for morphocompositional study, to obtain data about incidence and prevalence. On the other hand, findings about typology of stones and clinical and analytical characteristics of these subjects have been communicated in several meetings. Other valuable efforts in this field are the elaboration of guidelines for the collection and processing of urine samples for the study of urolithiasis in pediatric patients with the consensus of the Spanish Society for Pediatric Nephrology (AENP) as well as the Spanish Society for Laboratory Medicine (SEQC), the collaborative network RenalTube for the diagnosis of primary tubulopathies and the registry of patients with Primary Hyperoxaluria (OxalSpain). In many hospitals from the public healthcare system, pediatric nephrologists are the specialists in charge of the management of children with kidney stones, but there is no formal regulation on this competence. Other specialists, such as urologists, pediatric surgeons or pediatric urologists, in many cases do not offer a complete insight into the etiopathogenic mechanisms and the consequent medical treatment. Access to medication according to standards of treatment is warranted, provided a correct diagnosis is achieved, but criteria for the reimbursement of certain therapies, such as RNAi drugs for primary hyperoxaluria, are arguable.

Synergistic depolymerisation of alginate and chitosan by high hydrostatic pressure (HHP) and pulsed electric fields (PEF) treatment in the presence of H2O2.

Physically-induced depolymerisation procedures are often preferred for obtaining alginate and chitosan oligosaccharides as they either do not use or make minimal use of additional chemicals; therefore, separation of the final products is facile. In this work, solutions of three types of alginate with different mannuronic and guluronic acid residues ratio (M/G ratio) and molecular weights (Mw) and one type of chitosan were non-thermally processed by applying high hydrostatic pressures (HHP) up to 500 MPa (20 min) or pulsed electric fields (PEF) up to 25 kV cm-1 (4000 µm) in the absence or presence of 3 % hydrogen peroxide (H2O2). The impact on the physicochemical properties of alginate and chitosan was investigated by rheology, GPC, XRD, FTIR, and 1H NMR. In the rheological investigations, the apparent viscosities of all samples decreased with increasing shear rate, indicating a non-Newtonian shear-thinning behaviour. GPC results reported Mw reductions that ranged between 8 and 96 % for all treatments. NMR results revealed that HHP and PEF treatment predominantly reduced the M/G ratio of alginate and the degree of deacetylation (DDA) of chitosan, whilst H2O2 promoted an increase in the M/G ratio in alginate and DDA of chitosan. Overall, the present investigation has demonstrated the feasibility of HHP and PEF for rapidly producing alginate and chitosan oligosaccharides.

Structural and functional properties of alginate obtained by means of high hydrostatic pressure-assisted extraction.

The effects of the high hydrostatic pressure (HPP) pre-treatment on the alginate extraction were seen to greatly depend on the recalcitrant nature of two algae species. Alginates were deeply characterized in terms of composition, structure (HPAEC-PAD, FTIR, NMR, SEC-MALS), functional and technological properties. The pre-treatment significantly increased the alginate yield in the less recalcitrant A. nodosum (AHP) also favoring the extraction of sulphated fucoidan/fucan structures and polyphenols. Although the molecular weight was significantly lower in AHP samples, neither the M/G ratio nor the M and G sequences were modified. In contrast, a lower increase in alginate extraction yield was observed for the more recalcitrant S. latissima after the HPP pre-treatment (SHP), but it significantly affected the M/G values of the resulting extract. The gelling properties of the alginate extracts were also explored by external gelation in CaCl2 solutions. The mechanical strength and nanostructure of the hydrogel beads prepared were determined using compression tests, synchrotron small angle X-ray scattering (SAXS), and cryo-scanning electron microscopy (Cryo-SEM). Interestingly, the application of HPP significantly improved the gel strength of SHP, in agreement with the lower M/G values and the stiffer rod-like conformation obtained for these samples.

Gene expression study alerted to possible impairment in Daphnia magna individuals as a consequence of exposure to sublethal concentrations of prochloraz.

In the present study, Daphnia magna individuals were exposed for 21 days to 87, 130, 170, 230 and 380 µg/L of prochloraz. The expression of genes related to lipid metabolism (fabd), oxidative stress (cat and gst), heat shock proteins synthesis (hsp70 and hsp90), haemoglobin synthesis (hgb1 and hgb2), metallothioneins synthesis (mt-a, mt-b and mt-c), and vitellogenin synthesis (vgt1 y vgt2) were analyzed. Results showed that some gene expression in D. magna was altered as a consequence of the individual exposure to the fungicide. The genes fabd, vtg1 and vtg2, cat and gst resulted unaltered by the exposure of the daphnids to different fungicide concentrations. However, daphnid exposure to 380 µg/L of prochloraz resulted in an overexpression (p < 0.05) of hsp70 gene which indicated an alteration of the normal protein synthesis and its integrity maintenance. On the other hand, mt-b gene resulted significantly underexpressed (p < 0.05) in daphnids exposed to the lowest fungicide concentrations (87, 130 and 170 µg/L, respectively). In addition, hgb1 and hgb2 genes which are related with the haemoglobin synthesis weresignificantly overexpressed (p < 0.05). Results showed that hgb1 gene was overexpressed almost 100 times more in daphnids exposed for 21 days to 170, 230 and 380 µg/L than control values. However, many authors advocate for an association of these gene expression changes with the presence of contaminants in the medium, in fact they could be used as a good indicator of early contamination at low concentrations of toxicants.

The prochloraz chronic exposure to Daphnia magna derived in biochemical alterations of F0 generation daphnids and malformed F1 progeny.

In the present study, D. magna individuals were exposed to several sublethal prochloraz concentrations (87, 130, 170, 230 and 380 µg/L) for 21 days according to; the previous acute toxicity results. The fungicide effects on reproduction, survival, individual size, and growth population rate were evaluated after an exposure of 21 days, and no changes were observed compared to the control group. On the other hand, F1 generation neonates were collected and their external morphology evaluated; to estimate if the fungicide concentrations used induced effects during oogenesis and; embryogenesis processes. Neonates from parents which were previously exposed to 170 µg/L and higher concentrations were malformed since 16-d of exposure onward. All animals presented the same malformation: asymmetrical shell morphology and separated valves that did not cover the complete animal body regardless of the; concentration. The biochemical parameters tested in the broodstock were cholesterol, triglycerides, glucose and LDH activity. At the end of the chronic exposure experiment, cholesterol and triglycerides remained unaltered while glucose and the LDH enzyme levels increased significantly. The results of the present work showed a direct effect of; prochloraz on D. magna individual growth, along with mobilization of some; biochemical intermediate metabolism. A daphnid stress response as a result of the fungicide presence in the medium could be an explanation for the metabolic disorders. On the other hand, the F1 malformed neonates found in the present study suggested an effect of prochloraz among different daphnid generations and more studies would be necessary in this field.

Renal Dysplasia and Precocious Diabetes Onset in Microcephalic Osteodysplastic Primordial Dwarfism Type II Syndrome: A Case Report.

Microcephalic osteodysplastic primordial dwarfism type II (MOPDII) is a genetic syndrome. Its main characteristics are bony dysplasia, prenatal and postnatal growth deficiencies, microcephaly, and cerebrovascular disease. Several other features have been added recently. We report an individual with MOPDII affected by congenital renal dysplasia and hyperosmolar coma diabetic onset. Renal dysplasia has not been previously described in individuals with MOPDII. By publishing cases of unusual genetic disorders, it will be possible to broaden the spectrum of these rare syndromes, and improve the diagnosis and management of comorbidities.

Brief Report: CYP27B1 rs10877012 T Allele Was Linked to Non-AIDS Progression in ART-Naïve HIV-Infected Patients: A Retrospective Study.

BACKGROUND: HIV/AIDS progression is linked to vitamin D, which is regulated by several key cytochromes P450 (CYP). Single nucleotide polymorphisms (SNPs) in CYP genes influence vitamin D metabolism and serum levels. The objective of this study was to evaluate the association between CYP SNPs and the clinical AIDS progression in antiretroviral treatment (ART)-naïve HIV-infected patients. METHODS: We performed a retrospective study in 661 ART-naïve HIV-infected patients who were stratified by their AIDS progression pattern [181 long-term nonprogressors (LTNPs), 332 moderate progressors, and 148 rapid progressors (RPs)]. Four CYP SNPs (CYP2R1 rs10500804, CYP2R1 rs1993116, CYP27B1 rs10877012, and CYP24A1 rs6013897) were genotyped using Agena Bioscience's MassARRAY platform. Correction for multiple testing was performed using the false discovery rate (Benjamini-Hochberg procedure). RESULTS: The adjusted regression showed a significant association only for CYP27B1 rs10877012 SNP. When analyzing all HIV patients, the rs10877012 T allele was protective against AIDS progression (ordinal outcome) under the dominant [adjusted odds ratio (aOR) = 0.69; P = 0.021) and additive (aOR) = 0.75; P = 0.025] inheritance models. When analyzing LTNPs versus RPs, the rs10877012 T allele also showed a significant protective association under the dominant (aOR = 0.45; P = 0.004) and additive (aOR = 0.54; P = 0.008) inheritance models. P values remained significant after correcting by multiple comparisons only for the comparison of LTNPs versus RPs (extreme phenotypes). CONCLUSIONS: The CYP27B1 rs10877012 T allele was linked to non-AIDS progression in ART-naïve HIV-infected patients. The rs10877012 SNP seems to have an impact on the clinical AIDS progression, possibly modifying vitamin D levels, which could be relevant for the pathogenesis of HIV infection.

Estudio experimental comparativo de tres vías de aplicación de la cefazolina en las endoftalmitis bacterianas del conejo / Experimental compartive study of three ways application of cephazoline in bacterial endophtalmitis of the rabbit

Tradicionalmente las endoftalmitis bacterianas han sido tratadas con base en antibioticos tópicos y sistémicos. En la mayoría de los casos, la administración de antibióticos sistémicos no es inocua pues produce insuficiencia renal, ototoxidad, enterocolitis membranosa, diátesis hemorrágicas, problemas anafilácticos, etcétera. En numerosos estudios se ha demostrado que los antibióticos aplicados sistémicamente no alcanzan concentraciones adecuadas a nivel intraocular, por lo cual no es conveniente su aplicación en los casos de endoftalmitis exógenas bacterianas cuando no exista peligro de diseminación sistémica. Material y método. A 30 conejos albinos se les inoculó Staphylococcus aureus coagulasa positiva en el vítreo. Se hicieron tres grupos de 10 conejos. Al primer grupo se le aplicó cefazolina por vía sistémica, el segundo cefazolina intravítrea y al tercero cefazolina tanto por vía sistémica como intravítrea. Los conejos fueron sacrificados a las 72 horas y se realizó el estudio macro y microscópico de los globos oculares. Resultados. Se demostró que la administración del antibiótico por vía sistémica es inútil y su empleo en combinación con vía intravítrea es innecesario. Conclusión. Se comprobó la utilidad de la administración del antibiótico por vía intravítrea