Frecuencia de factores asociados a ideación suicida en pacientes con fibromialgia en dos hospitales de Lambayeque 2015

Objetivo: determinar la frecuencia de los factores asociados a ideación suicida en pacientes con fibromialgia en dos hospitales de Lambayeque, 2015. Material y Métodos: estudio descriptivo transversal con análisis exploratorio. Se entrevistó a pacientes con fibromialgia de consultorios externos de los hospitales Regional Lambayeque (Hospital 1) y Almanzor Aguinaga Asenjo de Chiclayo (Hospital 2). Se aplicaron 5 cuestionarios: la escala del centro de estudios epidemiológicos para depresión, escala de Zung para ansiedad, índice de calidad de sueño de Pittsburgh, escala de valoración análoga del dolor y la escala de ideación suicida de Beck, adaptada a población Lambayecana. Se realizó análisis univariado y bivariado exploratorio. Resultados: Se entrevistaron 125 pacientes, 61 del hospital 1 y 64 del Hospital 2; 87%, fueron mujeres. Hubieron 73,6% malos dormidores, 68% tuvieron depresión, 22,4% ansiedad, 24,8% dolor corporal difuso moderado-severo y 30,4% ideación suicida, 27,9% y 32,8% en el primer y segundo hospital, respectivamente. En los pacientes con Ideación suicida la frecuencia fue: mala calidad del sueño 81,6% (p=0,1), depresión 71%(p=0,5), ansiedad 42% (0,001) y dolor corporal difuso (DCD) de moderado a severo 26,3% (p=0,005). En el multivariado, ansiedad, rp= 1,4 IC95%= 1,14-1,7, p= 0,006 y ser mal dormidor, rp= 1,2, IC95%= 1,0-1,4, p=0,01 se asociaron a Ideación suicida. Conclusiones: la frecuencia de ideación suicida es elevada en estos pacientes. El factor más frecuente fue mala calidad de sueño. Ansiedad y ser mal dormidor se asociaron a ideación suicida.Tesi

Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study

Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life

Image_1_Hortaea werneckii isolates exhibit different pathogenic potential in the invertebrate infection model Galleria mellonella.jpeg

Hortaea werneckii is a black yeast with a remarkable tolerance to salt. Most studies have been dedicated to understanding how H. werneckii adapts to hypersaline environments. H. werneckii has an unconventional cell cycle in which it alternates between fission and budding, which is modulated by cell density. Additionally, H. werneckii can cause superficial mycosis of the palm and sole of humans. Here, we determine the impact of salt concentration on the EXF-2000 strain’s cell division pattern and morphology by performing timelapse microscopy at different salt concentrations. At low density and no salt, EXF-2000 primarily grows as pseudohyphae dividing mainly by septation. When grown in the presence of salt at a similar concentration to saltwater or hypersaline environments, we observe it grows first by undergoing fission followed by budding at the poles. Then, we examined a collection of 16 isolates in the presence of 0.6M NaCl, including isolates from marine and hypersaline environments and isolates from patients. These isolates exhibit a wide diversity in colony shape and cellular morphology. The isolates grew as yeast, pseudohyphae, and true hyphae, indicating that isolates can exhibit various cell morphologies under similar environmental conditions. We used the insect larvae Galleria mellonella to determine the pathogenic potential of our isolates. We observe that only a subset of isolates can cause death in our model, and there was no correlation between H. werneckii morphology and capacity to cause disease. Taken together, H. werneckii genomic and phenotypic diversity can serve as a model to better understand how phenotypes and pathogenic potential evolve in environmental fungi.</p

Construction of the FKBP12 deletion strains.

<p><b>(A) Construction of Δ<i>fkbp12-1</i> strain</b>. In the Δ<i>fkbp12-1</i> strain, wild-type <i>A</i>. <i>fumigatus fkbp12-1</i> (637 bp) was replaced by the 3.0 kb <i>A</i>. <i>parasiticus pyrG</i> gene. Three of the five strains validated by PCR were then selected for Southern analyses. SalI-digested genomic DNA was probed with the 646 bp probe of the downstream flanking sequence to confirm homologous recombination. Two of the three tested strains demonstrated the expected ~3.3 kb length, which is contrasted with the WT length of ~1.1 kb. Gel used for Southern analysis was 1% agarose. <b>(B) Construction of Δ<i>fkbp12-2</i> strain.</b> In the Δ<i>fkbp12-2</i> strain, wild-type <i>A</i>. <i>fumigatus fkbp12-2</i> (709 bp) was replaced with the 3.0 kb <i>A</i>. <i>parasiticus pyrG</i> gene. The strain validated by PCR was then used for Southern analyses. BamHI-digested genomic DNA was probed with the 733 bp probe of the downstream flanking sequence to confirm homologous recombination. The strain demonstrated the expected ~1.1 kb length in contrast with the ~2 kb length in the wild-type strain. Gel used for Southern analysis was 1.5% agarose. <b>(C) Construction of Δ<i>fkbp12-3</i> strain.</b> In the Δ<i>fkbp12-3</i> strain, wild-type <i>A</i>. <i>fumigatus fkbp12-3</i> (485 bp) was replaced by the 3.0 kb <i>A</i>. <i>parasiticus pyrG</i> gene. Four of the strains validated by PCR were then selected for Southern analyses. SacI-digested genomic DNA was probed with the 446 bp probe of the downstream flanking sequence to confirm homologous recombination. All four tested strains demonstrated the expected ~3.9 kb length as opposed to the wild-type length of ~1.4 kb. Gel used for Southern analysis was 1% agarose. <b>(D) Construction of Δ<i>fkbp12-4</i> strain.</b> In the Δ<i>fkbp12-4</i> mutant, wild-type <i>A</i>. <i>fumigatus fkbp12-4</i> (1653 bp) was replaced by the 3.0 kb <i>A</i>. <i>parasiticus pyrG</i> gene. Four of the strains validated by PCR were then selected for Southern analyses. BamHI-digested genomic DNA was probed with the 677 bp probe of the downstream flanking sequence to confirm homologous recombination. All four tested strains demonstrated the expected ~4.5 kb length as opposed to the wild-type length of ~2.0 kb. Gel used for Southern analysis was 1% agarose. <b>(E) Construction of Δ<i>fkbp12-1</i>Δ<i>fkbp12-2</i> strain</b>. In the Δ<i>fkbp12-1</i>Δ<i>fkbp12-2</i> strain, wild-type <i>A</i>. <i>fumigatus fkbp12-2</i> (709 bp) is replaced by the 4.4 kb hygromycin B resistance cassette in the Δ<i>fkbp12-1</i> strain. Four of the strains validated by PCR were then selected for Southern analyses. BamHI-digested genomic DNA was probed with the 550 bp probe of the downstream flanking sequence to confirm homologous recombination. All four tested strains demonstrated the expected ~5.2 kb as opposed to the wild-type length of ~1.9 kb.</p

Δ<i>fkbp12-1</i> is resistant to FK506 but its response to other antifungal agents is unchanged.

<p><b>(A)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM at 37°C for 5 days. <b>(B)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM + 100 ng/mL FK506 at 37°C for 5 days. <b>(C)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM + 10 μg/mL cyclosporine A (CsA) at 37°C for 5 days. <b>(D)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM + 1 μg/mL caspofungin (CSP) at 37°C for 5 days <b>(E)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM + 1 μg/mL caspofungin at 37°C for 5 days <b>(F)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴ conidia) incubated on GMM + 100 ng/mL FK506 + 1 μg/mL caspofungin at 37°C for 5 days.</p

Phylogenetic analysis of FKBP12 proteins and multiple sequence alignment comparing residues important for FK506-FKBP12 binding.

<p><b>(A)</b> Phylogram showing orthologous FKBP12 proteins from Human (HsFKBP12), <i>S</i>. <i>cerevisiae</i> (ScFKBP12), <i>C</i>. <i>neoformans</i> (CnFKBP12), <i>M</i>. <i>circinelloides</i> (McFKBP12) and <i>A</i>. <i>fumigatus</i> (AfFKBP12-1, AfFKBP12-2, AfFKBP12-3 and AfFKBP12-4). Phylogenic analysis was performed using the respective amino acid sequences aligned with MUSCLE (v3.8.31) implemented in the PhyML program (v3.1/3.0 aLRT). Graphical representation of the phylogenetic tree was performed with TreeDyn (v198.3). <i>A</i>. <i>fumigatus</i> FKBP12 proteins are designated as AfFKBP12-1, AfFKBP12-2, AfFKBP12-3 and AfFKBP12-4, respectively. <b>(B)</b> 14 residues are known to be important for binding FKBP12 to FK506. Residues distinct from human FKBP12 are highlighted in a different color for each <i>A</i>. <i>fumigatus</i> FKBP12 and for each species-specific FKBP12 homolog.</p

FKBP12-4 is required for complete hyphal growth.

<p><b>(A)</b> Growth of <i>A</i>. <i>fumigatus</i> (10⁴ conidia) on GMM at 37°C for 5 days, with colony diameter measured every 24 hours, revealed no statistically significant difference in growth among Δ<i>fkbp12-1</i>, Δ<i>fkbp12-2</i>, Δ<i>fkbp12-1</i>Δ<i>fkbp12-2</i>, Δ<i>fkbp12-3</i> and wild-type strains. Δ<i>fkbp12-4</i> demonstrated reduced growth rate across all 5 days (p = 0.0161). <b>(B)</b> After the 5 day growth period, Δ<i>fkbp12-4</i> demonstrated reduced growth compared to wild-type strain but no other obvious phenotypic abnormalities were noted.</p

Δ<i>fkbp12-1</i> demonstrates full hyphal growth in response to FK506; Δ<i>fkbp12-4</i> is slightly tolerant to FK506.

<p><b>(A)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴/mL) incubated in RPMI for 24 hours. <b>(B)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴/mL) incubated in RPMI with 100 ng/mL FK506 for 24 hours. <b>(C)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴/mL) incubated in RPMI for 48 hours. <b>(D)</b><i>A</i>. <i>fumigatus</i> conidia (10⁴/mL) incubated in RPMI with 100 ng/mL FK506 for 48 hours.</p

Deletion of FKBP12 encoding genes did not alter virulence of <i>A</i>. <i>fumigatus</i>.

<p>Larvae of the wax moth <i>Galleria mellonella</i> were injected with 5 μl of 1 x 10⁸ spores/ml (a total inoculum of 2 x 10⁵ spores) of the wild type, Δ<i>fkbp12-1</i>, Δ<i>fkbp12-2</i>, Δ<i>fkbp12-3</i>, Δ<i>fkbp12-4</i>, and<i>Δfkbp12-1Δfkbp12-2</i> strains. 20 larvae were included in each arm. Infected larvae were incubated at 37°C with survival scored daily for 5 days.</p

FK506 altered the localization of FKBP12-1 to the hyphal septum.

<p><b>(A)</b> Functionality of the expressed FKBP12-1-EGFP was assessed by comparing the growth of the FKBP12-1-EGFP expression strain with the <i>akuB</i>^<i>KU80</i> strain either in the absence or presence of FK506 (0.1 μg/mL) <b>(B, C)</b> Under normal growth conditions, FKBP12-1 evenly distributes throughout the cytoplasm and is also found in the nucleus at the hyphal tips (panel B) and sub-apical compartment (panel C and panel D) (marked by a white arrow heads in panel B and by red arrowheads in panel C and panel D). It is not seen at the septum (marked by a white arrow in the Fig 7C inset). <b>(C)</b> In the presence of FK506, FKBP12-1 can be seen localized as a double bar on either side of the septa indicating its binding to calcineurin complex at the hyphal septum (marked by a white arrows in the Fig 7D inset).</p