Ultraestructura dels espermatozoides de Crassostrea gigas, Mytilus galloprovincialis, Donax trunculus (Mollusca,Bivalvia)

La majoria dels mol·luscs bivalves són organismes gonocòrics, amb alguns casos d'hermafroditisme en diferents grups. La gònada masculina, quan és madura, envaeix tota la cavitat del mantell i els gàmetes són alliberats a la cavitat paleal de l'individu. Els espermatozoides dels bivalves són del tipus primitiu, com tots els dels organismes aquàtics que presenten fecundació externa. Són cèl·lules en les quals es distingeixen tres parts ben diferenciades: cap, peça intermèdia i cua. En les tres espècies estudiades, Crassostrea gigas, Mytilus galloprovincialis i Donax trunculus, els espermatozoides responen a aquest patró, però amb variacions morfològiques especifiques que estan relacionades amb les característiques dels oòcits que han de fecundar. En C. gigas el cap de l'espermatozoide és arrodonit i entre l'acrosoma i el nucli destaca la regió subacrosòmica, amb components fibril·lars d'actina. L'espermatozoide de M. galloprovincialis és molt més allargat que l'anterior i el sistema fibril·lar de la regió subacrosòmica forma un canal intranuclear. L'acrosoma destaca per la seva longitud. L'espermatozoide de D. trunculus segueix el patró considerat primitiu, tot i que en la peça intermèdia conté més mitocondris i els centríols tenen una disposició en línia i no ortogonal, com en els altres estudiats

Association of Thymidylate Synthase Polymorphisms with Acute Pancreatitis and/or Peripheral Neuropathy in HIV-Infected Patients on Stavudine-Based Therapy

Altres ajuts: Fondo de Investigaciones Sanitarias (FIS 02/1280, 05/1591, 07/0976, 08/00256), Fundación para la Prevención del SIDA en España (FIPSE 36610, 36572/06), and Red de Investigación en SIDA (RIS RD06/006/0022)Low expression thymidylate synthase (TS) polymorphism has been associated with increased stavudine triphosphate intracellular (d4T-TP) levels and the lipodystrophy syndrome. The use of d4T has been associated with acute pancreatitis and peripheral neuropathy. However, no relationship has ever been proved between TS polymorphisms and pancreatitis and/or peripheral neuropathy. We performed a case-control study to assess the relationship of TS and methylene-tetrahydrofolate reductase (MTHFR) gene polymorphisms with acute pancreatitis and/or peripheral neuropathy in patients exposed to d4T. Student's t test, Pearson's correlations, one-way ANOVA with Bonferroni correction and stepwise logistic regression analyses were done. Forty-three cases and 129 controls were studied. Eight patients (18.6%) had acute pancreatitis, and 35 (81.4%) had peripheral neuropathy. Prior AIDS was more frequent in cases than in controls (OR = 2.36; 95%CI 1.10-5.07, P = 0.0247). L7ow expression TS and MTHFR genotype associated with increased activity were more frequent in patients with acute pancreatitis and/or peripheral neuropathy than in controls (72.1% vs. 46.5%, OR = 2.97; 95%CI: 1.33-6.90, P = 0.0062, and 79.1% vs. 56.6%, OR = 2.90, 95%CI: 1.23-7.41, P = 0.0142, respectively). Independent positive or negative predictors for the development of d4T-associated pancreatitis and/or peripheral neuropathy were: combined TS and MTHFR genotypes (reference: A+A; P = 0.002; OR = 0.34 [95%CI: 0.08 to 1.44], OR = 3.38 [95%CI: 1.33 to 8.57], OR = 1.13 [95%CI: 0.34 to 3.71]), nadir CD4 cell count >200 cells/mm 3 (OR = 0.38; 95%CI: 0.17-0.86, P = 0.021), and HALS (OR = 0.39 95%CI: 0.18-0.85, P = 0.018). Low expression TS plus a MTHFR genotype associated with increased activity is associated with the development of peripheral neuropathy in d4T-exposed patients

Differential effects of dolutegravir, bictegravir and raltegravir in adipokines and inflammation markers on human adipocytes

Altres ajuts: European Regional Development Fund (FEDER); Gilead. European Union NextGeneration EU/PRTR.Aims: To assess the potential direct effects of the integrase strand-transfer inhibitors (INsTIs) dolutegravir, bictegravir, and raltegravir, drugs used as treatment for people living with human immunodeficiency virus (PLWH), on human adipose cells. Main methods: Drugs were added to the differentiation medium of human Simpson-Golabi-Behmel syndrome (SGBS) adipose cells and morphological adipogenesis was monitored for 10 days. Also, adipocytes were exposed to drugs following differentiation (day 14). The gene expression levels of selected adipogenesis markers, adipocyte metabolism markers, adipokines, and cytokines were determined by quantitative-reverse transcription polymerase-chain reaction. The release of adiponectin and leptin into the culture medium was measured using specific enzyme-linked immunosorbent assay, and release of interleukin-6 and chemokine (C[sbnd]C motif) ligand-2 using Multiplex assays. Key findings: Overall morphological adipogenesis was unaltered by INsTIs. The expression of adipogenesis marker genes (peroxisome proliferator-activated receptor-Ɣ and lipoprotein lipase) was slightly reduced in dolutegravir-treated differentiating adipocytes. Bictegravir repressed gene expression and the release of pro-inflammatory cytokines in differentiating adipocytes. Dolutegravir and raltegravir increased interleukin-6 gene expression, but only dolutegravir increased interleukin-6 release. Dolutegravir repressed adiponectin expression and release in differentiating adipocytes and had a similar but milder effect on leptin. Drug treatment of mature adipocytes reduced adiponectin gene expression in response to dolutegravir. Significance: The INsTIs studied do not have a significant effect on human adipose cell differentiation but exert distinct effects on gene expression and secretion of adipokines and cytokines. These findings will help understand and manage the effects of INsTI-containing treatments on body weight and metabolic dysregulation in PLWH

The impact of the COVID-19 pandemic in diabetes and dyslipidemia management in a Spanish region: a retrospective study of the Aragon population

IntroductionPrevious research has indicated that the COVID-19 outbreak had a negative impact on the diagnosis and management of cardiometabolic diseases. Our aim was to analyze the impact of the COVID-19 pandemic on the management of dyslipidemia and type 2 diabetes (T2D) in the Aragon region of Spain.MethodsWe conducted an observational retrospective study, which included data from all patients diagnosed with active T2D or dyslipidemia in Aragon during 2019–2021. Data was collected from the BIGAN platform, a big database that includes all healthcare data from the Aragon population. Clinical, biochemical, and pharmacological prescription information was obtained for each patient and for each year.ResultsOut of the total population of 1,330,000 in the Aragon region, 90,000 subjects were diagnosed with T2D each year, resulting in a prevalence of approximately 7%. The COVID-19 pandemic resulted in a decrease in the prevalence of this disease and a lower incidence during the year 2020. In addition, patients with T2D experienced a deterioration of their glucose profile, which led to an increase in the number of patients requiring pharmacological therapy. The prevalence of dyslipidemia was approximately 23.5% in both 2019 and 2020 and increased to 24.5% in 2021. Despite the worsening of the anthropometric profile, the lipid profile improved significantly throughout 2020 and 2021 compared to 2019. Moreover, the number of active pharmacological prescriptions increased significantly in 2021.DiscussionOur findings suggest that the overload of the health system caused by the COVID-19 pandemic has resulted in an underdiagnosis of T2D. Moreover, patients with T2D experienced a worsening of their glycemic profile, an increase in their pharmacological requirements, and lower performance of their analytical determinations. Dyslipidemic subjects improved their lipid profile although the value of lipid profile determination decreased between 2020 and 2021

Differential effects of dolutegravir, bictegravir and raltegravir in adipokines and inflammation markers on human adipocytes.

Aims: To assess the potential direct effects of the integrase strand-transfer inhibitors (INsTIs) dolutegravir, bictegravir, and raltegravir, drugs used as treatment for people living with human immunodeficiency virus (PLWH), on human adipose cells. Main methods: Drugs were added to the differentiation medium of human Simpson-Golabi-Behmel syndrome (SGBS) adipose cells and morphological adipogenesis was monitored for 10 days. Also, adipocytes were exposed to drugs following differentiation (day 14). The gene expression levels of selected adipogenesis markers, adipocyte metabolism markers, adipokines, and cytokines were determined by quantitative-reverse transcription polymerase-chain reaction. The release of adiponectin and leptin into the culture medium was measured using specific enzyme-linked immunosorbent assay, and release of interleukin-6 and chemokine (CC motif) ligand-2 using Multiplex assays. Key findings: Overall morphological adipogenesis was unaltered by INsTIs. The expression of adipogenesis marker genes (peroxisome proliferator-activated receptor-Ɣ and lipoprotein lipase) was slightly reduced in dolutegravir-treated differentiating adipocytes. Bictegravir repressed gene expression and the release of pro-inflammatory cytokines in differentiating adipocytes. Dolutegravir and raltegravir increased interleukin-6 gene expression, but only dolutegravir increased interleukin-6 release. Dolutegravir repressed adiponectin expression and release in differentiating adipocytes and had a similar but milder effect on leptin. Drug treatment of mature adipocytes reduced adiponectin gene expression in response to dolutegravir. Significance: The INsTIs studied do not have a significant effect on human adipose cell differentiation but exert distinct effects on gene expression and secretion of adipokines and cytokines. These findings will help understand and manage the effects of INsTI-containing treatments on body weight and metabolic dysregulation in PLWH

Association of thymidylate synthase polymorphisms with acute pancreatitis and/or peripheral neuropathy in HIV-infected patients on Stavudine-Based Therapy

BACKGROUND: Low expression thymidylate synthase (TS) polymorphism has been associated with increased stavudine triphosphate intracellular (d4T-TP) levels and the lipodystrophy syndrome. The use of d4T has been associated with acute pancreatitis and peripheral neuropathy. However, no relationship has ever been proved between TS polymorphisms and pancreatitis and/or peripheral neuropathy. METHODS: We performed a case-control study to assess the relationship of TS and methylene-tetrahydrofolate reductase (MTHFR) gene polymorphisms with acute pancreatitis and/or peripheral neuropathy in patients exposed to d4T. Student's t test, Pearson's correlations, one-way ANOVA with Bonferroni correction and stepwise logistic regression analyses were done. RESULTS: Forty-three cases and 129 controls were studied. Eight patients (18.6%) had acute pancreatitis, and 35 (81.4%) had peripheral neuropathy. Prior AIDS was more frequent in cases than in controls (OR = 2.36; 95%CI 1.10-5.07, P = 0.0247). L7ow expression TS and MTHFR genotype associated with increased activity were more frequent in patients with acute pancreatitis and/or peripheral neuropathy than in controls (72.1% vs. 46.5%, OR = 2.97; 95%CI: 1.33-6.90, P = 0.0062, and 79.1% vs. 56.6%, OR = 2.90, 95%CI: 1.23-7.41, P = 0.0142, respectively). Independent positive or negative predictors for the development of d4T-associated pancreatitis and/or peripheral neuropathy were: combined TS and MTHFR genotypes (reference: A+A; P = 0.002; ORA+B = 0.34 [95%CI: 0.08 to 1.44], ORB+A = 3.38 [95%CI: 1.33 to 8.57], ORB+B = 1.13 [95%CI: 0.34 to 3.71]), nadir CD4 cell count >200 cells/mm(3) (OR = 0.38; 95%CI: 0.17-0.86, P = 0.021), and HALS (OR = 0.39 95%CI: 0.18-0.85, P = 0.018). CONCLUSIONS: Low expression TS plus a MTHFR genotype associated with increased activity is associated with the development of peripheral neuropathy in d4T-exposed patients

Atlas de histopatología de moluscos bivalvos. Introducción a la histología de algunas especies de moluscos bivalvos marinos y sus parasitosis más frecuentes

Maquetació: Pablo Bou Mira[cat] Els exemplars de mol·luscs bivalves estudiats procedeixen principalment del Delta de l´Ebre (badia dels Alfacs i del Fangar) i les ostres han estat de dues procedències diferents: Delta de l´Ebre i de Sète (França). S´han estudiat individus immadurs (juvenils) i s´ha seguit el procés de maduració sexual en tots els casos. Les tècniques aplicades en aquest estudi han seguit els protocols estàndards. Per la Microscòpia Òptica: fixació en formol al 10% o en Bouin. Inclusió en parafina i obtenció de talls de cinc a set micres per terme mig. Les tincions rutinàries han estat post vitals amb hematoxilina-eosina (Hem-Eos) y també amb el tricròmic de Mallory. El taronja d´acridina s´ha emprat per les observacions amb el microscopi de fluorescència i el iodur de propidi-fal·loïdina-TRITC (isocianat de tetrametilrodamina) per les observacions amb el microscopi confocal i en ocasions també s´ha utilitzat la doble tinció hematoxilina-eosina (Hem-Eos). En alguns casos s´han fet tincions vitals amb roig neutre i amb blau de metilè. Per l´estudi amb la Microscòpia Electrònica de Rastreig (MER) la fixació ha estat en glutaraldehid-paraformaldehid preparat amb tampó de Sörensen o amb cacodilat i l´ombrejat amb carbó i també s´ha metal·litzat amb or o amb una aleació d´or-pal·ladi. Per la Microscòpia Electrònica de Transmissió (MET) la fixació ha estat doble: glutaraldehid-paraformaldehid preparat amb la solució de Sörensen (a base de fosfats) o amb cacodilat al 0,1 M seguit d´una postfixació, generalment amb tetraòxid d´osmi a l´1% igualment tamponat amb la mateixa solució emprada en la primera fixació. La inclusió ha estat preferentment feta amb Spurr i els talls semifins (de control) tenyits amb blau de metilè-bòrax i els ultrafins contrastats amb acetat d´uranil i citrat de plom. Les observacions s´han fet amb microscopis electrònics Leica, Hitachi, Jeol, Philips y Zeiss dels Centres Científics i Tecnològics de la Universitat de Barcelona.[spa] Los ejemplares de moluscos bivalvos estudiados proceden principalmente del Delta del Ebro (bahía de los Alfacs i del Fangar) y las ostres tienen dos procedencias: Delta del Ebro y Sète (Francia). Se han procesado ejemplares inmaduros (juveniles) y se ha seguido el proceso de maduración sexual. Las técnicas aplicadas en este estudio han seguido los protocolos estándar. Para la Microscopía Òptica: fijación en formol al 10% o en Bouin. Inclusión en parafina y obtención de cortes finos de unas cinco a siete micras de grosor por término medio. Las tinciones rutinarias han sido preferentemente las postvitales con hematoxilina-eosina y también con el tricrómico de Mallory. Ocasionalmente se ha utilizado en naranja de acridina para las observaciones con el microscopio de fluorescencia. Para las observaciones con el microscopio confocal se ha empleado el ioduro de propidio-faloidina-TRITC (isocianato de tetrametilrodamina) y también la doble tinción hematoxilina-eosina (Hem-Eos). En algunos casos se han efectuado tinciones vitales con rojo neutro y con azul de metileno. Para el estudio con el Microscopio Electrónico de Barrido (MEB) la fijación ha sido con glutaraldehido- paraformaldehido preparado con tampón fosfato de Sörensen o con cacodilato sódico 0,1 M a pH 7,3. El sombreado se ha hecho con carbón y en ocasiones también se ha metalizado con oro o con una aleación de oro-paladio. Para la Microscopía Electrónica de Transmisión (MET) se ha procedido a la fijación con glutaraldehido- paraformaldehido preparado con la solución tampón Sörensen (a base de fosfatos) o con cacodilato sódico al 0,1 M, seguido de una postfijación con tetraóxido de osmio al 1% igualmente preparado con el tampón utilizado en la primera fijación. La inclusión se ha hecho preferentemente con Spurr y los cortes semifinos (de control) se han teñido con azul de metileno-bórax y los ultrafinos han sido contrastados con acetato de uranilo y citrato de plomo. Las observaciones se han efectuado con microscopios electrónicos Leica, Philips, Hitachi, Jeol, Philips y Zeiss de los Centros Científicos y Tecnológicos de la Universidad de Barcelona

Effects of Switching from Stavudine to Raltegravir on Subcutaneous Adipose Tissue in HIV-Infected Patients with HIV/HAART-Associated Lipodystrophy Syndrome (HALS). A Clinical and Molecular Study

HIV-1/HAART-associated lipodystrophy syndrome (HALS) has been associated with exposure to stavudine (d4T) through mitochondrial dysfunction. We performed a 48-week study to assess the effects of switching from d4T to raltegravir (RAL) on metabolic and fat molecular parameters of patients with HALS. Forty-two patients with HALS and a median exposure to d4T > 7 years were switched to RAL and followed for 48 weeks. Fasting metabolic tests, HIV RNA, CD4 cell count, and fat measured by DEXA were obtained at baseline and week 48. mtDNA and gene transcripts for PPAR gamma, adiponectin, cytochrome b, Cox IV, TNF alpha, MCP-1 and CD68 were assessed in paired subcutaneous fat tissue biopsies. Lipid parameters, fasting glucose, insulin, and HOMA-IR did not change significantly. Whole body fat (P = 0.0027) and limb fat mass (P<0.0001) increased from baseline. Trunk/limb fat ratio (P = 0.0022), fat mass ratio (P = 0.0020), fat mass index (P = 0.0011) and percent leg fat normalized to BMI (P<0.0001) improved after 48 weeks. Relative abundance of mtDNA, expression of PPAR gamma, adiponectin, Cyt b, and MCP-1 genes increased, whereas Cox IV, TNF alpha, and CD68 did not change significantly from baseline. Switching from d4T to RAL in patients with HALS is associated with an increase in limb fat mass and an improvement in markers of adipocyte differentiation and mitochondrial function in SAT

Polymorphisms of Pyrimidine Pathway Enzymes Encoding Genes and HLA-B*40∶01 Carriage in Stavudine-Associated Lipodystrophy in HIV-Infected Patients.

Purpose To assess in a cohort of Caucasian patients exposed to stavudine (d4T) the association of polymorphisms in pyrimidine pathway enzymes and HLA-B*40∶01 carriage with HIV/Highly active antiretroviral therapy (HAART)-associated lipodystrophy syndrome (HALS). Methods Three-hundred and thirty-six patients, 187 with HALS and 149 without HALS, and 72 uninfected subjects were recruited. The diagnosis of HALS was performed following the criteria of the Lipodystrophy Severity Grading Scale. Polymorphisms in the thymidylate synthase (TS) and methylene-tetrahydrofolate reductase (MTHFR) genes were determined by direct sequencing, HLA-B genotyping by PCR-SSOr Luminex Technology, and intracellular levels of stavudine triphosphate (d4T-TP) by a LC-MS/MS assay method. Results HALS was associated with the presence of a low expression TS genotype polymorphism (64.7% vs. 42.9%, OR = 2.43; 95%CI: 1.53-3.88, P<0.0001). MTHFR gene polymorphisms and HLA-B*40∶01 carriage were not associated with HALS or d4T-TP intracellular levels. Low and high expression TS polymorphisms had different d4T-TP intracellular levels (25.60 vs. 13.60 fmol/106 cells, P<0.0001). Independent factors associated with HALS were(OR [95%CI]: (a) Combined TS and MTHFR genotypes (p = 0.006, reference category (ref.): 'A+A'; OR for 'A+B' vs. ref.: 1.39 [0.69-2.80]; OR for 'B+A' vs. ref.: 2.16 [1.22-3.83]; OR for 'B+B' vs. ref.: 3.13, 95%CI: 1.54-6.35), (b) maximum viral load ≥5 log10 (OR: 2.55, 95%CI: 1.56-4.14, P = 0.001), (c) use of EFV (1.10 [1.00-1.21], P = 0.008, per year of use). Conclusion HALS is associated with combined low-expression TS and MTHFR associated with high activity polymorphisms but not with HLA-B*40∶01 carriage in Caucasian patients with long-term exposure to stavudine

Assessing Associations between the AURKA-HMMR-TPX2-TUBG1 Functional Module and Breast Cancer Risk in BRCA1/2 Mutation Carriers

While interplay between BRCA1 and AURKA-RHAMM-TPX2-TUBG1 regulates mammary epithelial polarization, common genetic variation in HMMR (gene product RHAMM) may be associated with risk of breast cancer in BRCA1 mutation carriers. Following on these observations, we further assessed the link between the AURKA-HMMR-TPX2-TUBG1 functional module and risk of breast cancer in BRCA1 or BRCA2 mutation carriers. Forty-one single nucleotide polymorphisms (SNPs) were genotyped in 15,252 BRCA1 and 8,211 BRCA2 mutation carriers and subsequently analyzed using a retrospective likelihood approach. The association of HMMR rs299290 with breast cancer risk in BRCA1 mutation carriers was confirmed: per-allele hazard ratio (HR) = 1.10, 95% confidence interval (CI) 1.04-1.15, p = 1.9 x 10(-4) (false discovery rate (FDR)-adjusted p = 0.043). Variation in CSTF1, located next to AURKA, was also found to be associated with breast cancer risk in BRCA2 mutation carriers: rs2426618 per-allele HR = 1.10, 95% CI 1.03-1.16, p = 0.005 (FDR-adjusted p = 0.045). Assessment of pairwise interactions provided suggestions (FDR-adjusted pinteraction values > 0.05) for deviations from the multiplicative model for rs299290 and CSTF1 rs6064391, and rs299290 and TUBG1 rs11649877 in both BRCA1 and BRCA2 mutation carriers. Following these suggestions, the expression of HMMR and AURKA or TUBG1 in sporadic breast tumors was found to potentially interact, influencing patients' survival. Together, the results of this study support the hypothesis of a causative link between altered function of AURKA-HMMR-TPX2-TUBG1 and breast carcinogenesis in BRCA1/2 mutation carriers