Brain and cognitive changes in patients with long COVID compared with infection-recovered control subjects.

Between 2.5 and 28% of people infected with SARS-CoV-2 suffer Long COVID or persistence of symptoms for months after acute illness. Many symptoms are neurological, but the brain changes underlying the neuropsychological impairments remain unclear. This study aimed to provide a detailed description of the cognitive profile, the pattern of brain alterations in Long COVID and the potential association between them. To address these objectives, 83 patients with persistent neurological symptoms after COVID-19 were recruited, and 22 now healthy controls chosen because they had suffered COVID-19 but did not experience persistent neurological symptoms. Patients and controls were matched for age, sex and educational level. All participants were assessed by clinical interview, comprehensive standardized neuropsychological tests and structural MRI. The mean global cognitive function of patients with Long COVID assessed by ACE III screening test (Overall Cognitive level - OCLz= -0.39± 0.12) was significantly below the infection recovered-controls (OCLz= +0.32± 0.16, p< 0.01). We observed that 48% of patients with Long COVID had episodic memory deficit, with 27% also impaired overall cognitive function, especially attention, working memory, processing speed and verbal fluency. The MRI examination included grey matter morphometry and whole brain structural connectivity analysis. Compared to infection recovered controls, patients had thinner cortex in a specific cluster centred on the left posterior superior temporal gyrus. In addition, lower fractional anisotropy (FA) and higher radial diffusivity (RD) were observed in widespread areas of the patients' cerebral white matter relative to these controls. Correlations between cognitive status and brain abnormalities revealed a relationship between altered connectivity of white matter regions and impairments of episodic memory, overall cognitive function, attention and verbal fluency. This study shows that patients with neurological Long COVID suffer brain changes, especially in several white matter areas, and these are associated with impairments of specific cognitive functions.

Interventions, methods and outcome measures used in teaching evidence-based practice to healthcare students: an overview of systematic reviews.

BACKGROUND: To fully implement the internationally acknowledged requirements for teaching in evidence-based practice, and support the student's development of core competencies in evidence-based practice, educators at professional bachelor degree programs in healthcare need a systematic overview of evidence-based teaching and learning interventions. The purpose of this overview of systematic reviews was to summarize and synthesize the current evidence from systematic reviews on educational interventions being used by educators to teach evidence-based practice to professional bachelor-degree healthcare students and to identify the evidence-based practice-related learning outcomes used. METHODS: An overview of systematic reviews. Four databases (PubMed/Medline, CINAHL, ERIC and the Cochrane library) were searched from May 2013 to January 25th, 2024. Additional sources were checked for unpublished or ongoing systematic reviews. Eligibility criteria included systematic reviews of studies among undergraduate nursing, physiotherapist, occupational therapist, midwife, nutrition and health, and biomedical laboratory science students, evaluating educational interventions aimed at teaching evidence-based practice in classroom or clinical practice setting, or a combination. Two authors independently performed initial eligibility screening of title/abstracts. Four authors independently performed full-text screening and assessed the quality of selected systematic reviews using standardized instruments. Data was extracted and synthesized using a narrative approach. RESULTS: A total of 524 references were retrieved, and 6 systematic reviews (with a total of 39 primary studies) were included. Overlap between the systematic reviews was minimal. All the systematic reviews were of low methodological quality. Synthesis and analysis revealed a variety of teaching modalities and approaches. The outcomes were to some extent assessed in accordance with the Sicily group`s categories; "skills", "attitude" and "knowledge". Whereas "behaviors", "reaction to educational experience", "self-efficacy" and "benefits for the patient" were rarely used. CONCLUSIONS: Teaching evidence-based practice is widely used in undergraduate healthcare students and a variety of interventions are used and recognized. Not all categories of outcomes suggested by the Sicily group are used to evaluate outcomes of evidence-based practice teaching. There is a need for studies measuring the effect on outcomes in all the Sicily group categories, to enhance sustainability and transition of evidence-based practice competencies to the context of healthcare practice.

Study of the human hippocampal formation: a method for histological and magnetic resonance correlation in perinatal cases.

Little information is available on the magnetic resonance imaging (MRI) determination of the hippocampal formation (HF) during the perinatal period. However, this exploration is increasingly used, which requires defining visible HF landmarks on MRI images, validated through histological analysis. This study aims to provide a protocol to identify HF landmarks on MRI images, followed by histological validation through serial sections of the temporal lobe of the samples examined, to assess the longitudinal extent of the hippocampus during the perinatal period. We examined ex vivo MRI images from nine infant control brain samples. Histological validation of the hippocampal formation MRI images was obtained through serial sectioning and examination of Nissl-stained sections at 250 µm intervals along the entire length of the hippocampal formation. Up to six landmarks were identified both in MRI images and the serial histological sections. Proceeding in an anterior to posterior (rostrocaudal) direction, these were as follows: 1) the limen insulae (fronto-temporal junction); 2) the beginning of the amygdaloid complex; 3) the beginning of the lateral ventricle; 4) the caudal limit of the uncus, indicated by the start of the lateral geniculate nucleus (at the level of the gyrus intralimbicus); 5) the end of the lateral geniculate nucleus (beginning of the pulvinar); and 6) the beginning of the fornix. After histological validation of each of these landmarks, the full longitudinal length of the hippocampal formation and distances between landmarks were calculated. No statistically significant differences were found in total length or between landmarks. While the HF is anatomically organized at birth, its annotation is particularly challenging to perform. The histological validation of HF landmarks allows a better understanding of MRI images. The proposed protocol could be useful to assess MRI hippocampal quantification in children and possible variations due to different neurological diseases.

Born Too Early and Too Small: Higher Order Cognitive Function and Brain at Risk at Ages 8-16.

Prematurity presents a risk for higher order cognitive functions. Some of these deficits manifest later in development, when these functions are expected to mature. However, the causes and consequences of prematurity are still unclear. We conducted a longitudinal study to first identify clinical predictors of ultrasound brain abnormalities in 196 children born very preterm (VP; gestational age ≤32 weeks) and with very low birth weight (VLBW; birth weight ≤1500 g). At ages 8-16, the subset of VP-VLBW children without neurological findings (124) were invited for a neuropsychological assessment and an MRI scan (41 accepted). Of these, 29 met a rigorous criterion for MRI quality and an age, and gender-matched control group (n = 14) was included in this study. The key findings in the VP-VLBW neonates were: (a) 37% of the VP-VLBW neonates had ultrasound brain abnormalities; (b) gestational age and birth weight collectively with hospital course (i.e., days in hospital, neonatal intensive care, mechanical ventilation and with oxygen therapy, surgeries, and retinopathy of prematurity) predicted ultrasound brain abnormalities. At ages 8-16, VP-VLBW children showed: a) lower intelligent quotient (IQ) and executive function; b) decreased gray and white matter (WM) integrity; (c) IQ correlated negatively with cortical thickness in higher order processing cortical areas. In conclusion, our data indicate that facets of executive function and IQ are the most affected in VP-VLBW children likely due to altered higher order cortical areas and underlying WM.

Registro Nacional Argentino de Shock Cardiogénico (ReNa-SHOCK) / Argentine National Registry of Cardiogenic Shock (ReNa-SHOCK)

Introducción: El shock cardiogénico es una complicación grave del infarto agudo de miocardio y constituye una de sus principales causas de muerte, pese a lo cual la información en nuestro medio es limitada. Objetivo: Conocer las características clínicas, estrategias de tratamiento y evolución intrahospitalaria del shock cardiogénico en la Argentina. Material y métodos: Se realizó un registro prospectivo, multicéntrico de pacientes internados con shock cardiogénico en el contexto de los síndromes coronarios agudos con y sin elevación del segmento ST entre los años 2013 y 2015 en 64 centros de la Argentina. Resultados: Se incluyeron 165 pacientes, con una edad media de 66 (58-76,5) años; el 65% eran hombres. El 75% de los casos cursaban un síndrome coronario agudo con elevación del segmento ST. El 8,5% estuvieron asociados con complicaciones mecánicas y el 6,7% con compromiso del ventrículo derecho. El 56% presentaban shock cardiogénico al ingreso. Requirieron inotrópicos el 95%, asistencia respiratoria mecánica el 78%, catéter de Swan-Ganz el 44%, balón de contrapulsación intraaór-tico el 37%. El 84% de los síndromes coronarios agudos con elevación del segmento ST (104/124 pacientes) se reperfundieron. La mediana de tiempo desde el inicio de los síntomas al ingreso fue de 240 minutos (132-720). El 80% recibieron angioplastia primaria. La mortalidad intrahospitalaria global fue del 54%, sin diferencias entre los síndromes coronarios agudos con o sin elevación del segmento ST. Asimismo, no hubo diferencia en la frecuencia de eventos y uso de procedimientos entre los síndromes coronarios agudos con o sin elevación del segmento ST. Conclusiones: Las características del shock cardiogénico en la Argentina no difieren mucho de poblaciones de otras partes del mundo. La morbimortalidad es elevada a pesar de la utilización de las estrategias de tratamiento disponibles.

Revisión de estudios sobre calidad de vida relacionada con la salud en la enfermedad renal crónica avanzada en España / Review of studies on health related quality of life in patients with advanced chronic kidney disease in Spain

Antecedentes: La enfermedad renal crónica avanzada (ERCA) tiene un gran impacto sobre la calidad de vida relacionada con la salud (CVRS). Cada vez es más frecuente el uso de esta variable en estudios en nuestro medio, aunque no se dispone de una revisión global sobre cómo se ha estudiado en población con ERCA española. Objetivos: Ofrecer una visión contrastada de los instrumentos de evaluación de la CVRS más usados en la población española con ERCA, analizando además la calidad de vida percibida por esta población. Métodos: Se llevó a cabo una revisión de la literatura publicada sobre estudios realizados en España que hubieran empleado algún instrumento para medir la CVRS, genérico o específico, en pacientes con diferentes estadios de ERCA. Se excluyeron estudios en pacientes trasplantados renales cuando eran estudiados de forma independiente. La búsqueda se realizó en CINAHL, CUIDEN, DOCUMED, EMBASE, ERIC (USDE), IME, LILACS, MEDLINE, Nursin@ovid, PubMed, Scielo, Web of Science y TESEO. Resultados: Se han incluido en esta revisión 53 artículos publicados entre el año 1995 y el mes de mayo de 2014. La terapia sustitutiva renal es la variable con mayor frecuencia asociada al estudio de la CVRS, siendo la hemodiálisis la más estudiada. La mayoría de los estudios encontrados son transversales y el Short Form-36 Health Survey es el instrumento más usado. Conclusiones: La mayoría de los estudios muestra cómo la CVRS se ve afectada de forma importante en pacientes que reciben terapia sustitutiva renal. Estos resultados se muestran independientes del instrumento usado para medir la calidad de vida relacionada con la salud y de otras variables asociadas a lo largo de los distintos estudios. La CVRS ha sido analizada especialmente en pacientes en hemodiálisis, con diseños fundamentalmente observacionales y con el Short Form-36 Health Survey. Se necesitan más estudios que aborden aspectos como la CVRS en la etapa prediálisis, así como estudios con muestras más grandes y diseños longitudinales, analíticos o experimentales (AU)

Background: Advanced chronic kidney disease (ACKD) has a great impact on health-related quality of life (HRQL). The use of this variable in studies in our field is becoming more frequent, although there has been no comprehensive review of how Spaniards with ACKD are assessed. Aims: To offer a contrasted vision of the HRQL assessment tools that are most often used on Spanish ACKD population, also analysing how this population perceive their quality of life. Method: A review was carried out on literature published on studies undertaken in Spain that had used some kind of instrument, either generic or specific, in order to measure HRQL in patients with different stages of ACKD. Studies in kidney transplant patients were excluded when they were independently reviewed. The research was carried out in CINAHL, CUIDEN, DOCUMED, EMBASE, ERIC (USDE), IME, LILACS, MEDLINE, Nursin@ovid, PubMed, Scielo, Web of Science and TESEO. Results: 53 articles published between 1995 and May 2014 have been included in this review. Renal replacement therapy is the variable that is most often associated with the study of HRQL, with haemodialysis being the most studied. Most of the studies found are cross-sectional and the Short Form-36 Health Survey is the most used instrument. Conclusions: The majority of the studies show how HRQL is significantly affected in patients who receive renal replacement therapy. These results are independent from the instrument used to measure health-related quality of life and other associated variables throughout the various studies. HRQL has been particularly analysed in patients on haemodialysis, using mainly observational methods and the Short Form-36 Health Survey. There is a need for more studies that address aspects such as HRQL in the pre-dialysis phase, as well as studies with larger samples and longitudinal, analytical and experimental designs (AU)

Review of studies on health related quality of life in patients with advanced chronic kidney disease in Spain.

BACKGROUND: Advanced chronic kidney disease (ACKD) has a great impact on health-related quality of life (HRQL). The use of this variable in studies in our field is becoming more frequent, although there has been no comprehensive review of how Spaniards with ACKD are assessed. AIMS: To offer a contrasted vision of the HRQL assessment tools that are most often used on Spanish ACKD population, also analysing how this population perceive their quality of life. METHOD: A review was carried out on literature published on studies undertaken in Spain that had used some kind of instrument, either generic or specific, in order to measure HRQL in patients with different stages of ACKD. Studies in kidney transplant patients were excluded when they were independently reviewed. The research was carried out in CINAHL, CUIDEN, DOCUMED, EMBASE, ERIC (USDE), IME, LILACS, MEDLINE, Nursin@ovid, PubMed, Scielo, Web of Science and TESEO. RESULTS: 53 articles published between 1995 and May 2014 have been included in this review. Renal replacement therapy is the variable that is most often associated with the study of HRQL, with haemodialysis being the most studied. Most of the studies found are cross-sectional and the Short Form-36 Health Survey is the most used instrument. CONCLUSIONS: The majority of the studies show how HRQL is significantly affected in patients who receive renal replacement therapy. These results are independent from the instrument used to measure health-related quality of life and other associated variables throughout the various studies. HRQL has been particularly analysed in patients on haemodialysis, using mainly observational methods and the Short Form-36 Health Survey. There is a need for more studies that address aspects such as HRQL in the pre-dialysis phase, as well as studies with larger samples and longitudinal, analytical and experimental designs.

Keratin23 (KRT23) knockdown decreases proliferation and affects the DNA damage response of colon cancer cells.

Keratin 23 (KRT23) is strongly expressed in colon adenocarcinomas but absent in normal colon mucosa. Array based methylation profiling of 40 colon samples showed that the promoter of KRT23 was methylated in normal colon mucosa, while hypomethylated in most adenocarcinomas. Promoter methylation correlated with absent expression, while increased KRT23 expression in tumor samples correlated with promoter hypomethylation, as confirmed by bisulfite sequencing. Demethylation induced KRT23 expression in vitro. Expression profiling of shRNA mediated stable KRT23 knockdown in colon cancer cell lines showed that KRT23 depletion affected molecules of the cell cycle and DNA replication, recombination and repair. In vitro analyses confirmed that KRT23 depletion significantly decreased the cellular proliferation of SW948 and LS1034 cells and markedly decreased the expression of genes involved in DNA damage response, mainly molecules of the double strand break repair homologous recombination pathway. KRT23 knockdown decreased the transcript and protein expression of key molecules as e.g. MRE11A, E2F1, RAD51 and BRCA1. Knockdown of KRT23 rendered colon cancer cells more sensitive to irradiation and reduced proliferation of the KRT23 depleted cells compared to irradiated control cells.

Impact of incident comorbidity on functional loss in elderly chronic kidney disease patients undergoing hemodialysis.

The incidence of end stage renal disease in older persons has been increasing progressively over the last 10 years. Improved survival rates with renal replacement therapy are making this increased prevalence even more pronounced. The usual risks of morbidity and requirements for specialized care associated with older people increase dramatically when they have chronic kidney disease (CKD). It has been seen that the majority of patients in hemodialysis units are over the age of 60, and have significant co-morbidities. The relationship between older age, chronic disorders and functional dependence (FD) is well known. Accordingly, nursing care planning must be designed with this in mind. The aim of this study was to assess whether the comorbidity associated with CKD modifies FD in patients on hemodialysis. We undertook a prospective longitudinal cohort study of hemodialysis outpatients in Málaga, Spain, using the Barthel test to establish FD and the Charlson comorbidity index to quantify comorbidity. All health events were analyzed to select those study patients with incident comorbidity, understood as the appearance of a new disease that could modify the Charlson comorbidity index, and determine the change in FD. Multivariate linear regression showed that the best model for predicting functional loss was that which considered comorbidity adjusted for age, particularly when it occurred as a result of hospital admission, as it was shown to have an important predictive value for the onset of a decrease in functional dependency scores in patients with CKD.

Cathepsin E, maspin, Plk1, and survivin are promising prognostic protein markers for progression in non-muscle invasive bladder cancer.

Bladder cancer is a common cancer with particularly high recurrence after transurethral resection. In this study, we investigated the prognostic value of the protein expression of cathepsin E, maspin, polo-like kinase 1 (Plk1), and survivin in patients with stage Ta and T1 urothelial carcinomas. Transcripts from the four genes encoding these proteins were previously included in gene expression signatures for outcome prediction for Ta/T1 bladder cancer. We used three different tissue microarrays with 693 non-muscle invasive urothelial carcinomas from Danish, Swedish, and Spanish patient cohorts with long-term follow-up. Protein expression was measured by immunohistochemistry, and antibody specificity was validated by Western blotting. In the Danish patient cohort, we found the expression of cathepsin E, maspin, Plk1, and survivin to be significantly associated with progression to stage T2 to T4 bladder cancer (for each marker: log-rank test; P < 0.001). Multivariate Cox regression analysis identified cathepsin E (P < 0.001), Plk1 (P = 0.021), maspin (P = 0.001), and survivin (P = 0.001) as independent prognostic markers. Furthermore, maspin, survivin, and cathepsin E expression significantly subgrouped patients already stratified by European Organization for Research and Treatment of Cancer risk scores. Finally, we successfully validated the results in tumors from 410 patients from both Sweden and Spain. We conclude that all four protein markers may have prognostic value in non-muscle invasive bladder cancer for guiding optimal treatment of patients. Additional prospective studies are needed for further validation of the clinical relevance of this marker panel.

Downregulation of zinc finger protein 132 in prostate cancer is associated with aberrant promoter hypermethylation and poor prognosis.

This study investigates the expression and biomarker potential of zinc finger protein 132 (ZNF132) in prostate cancer (PC) by transcriptional profiling and immunohistochemical analysis of tissue microarrays, including tumor specimens from 615 radical prostatectomy (RP) patients and 199 conservatively treated patients. Primary clinical endpoints were time to PSA recurrence and cancer-specific death, respectively. Compared to normal prostate epithelial cells from men without PC, ZNF132 transcript levels were significantly reduced in PC cells from patients with localized PC and further downregulated in metastatic PC. Likewise, ZNF132 protein expression was significantly lower in primary tumors from patients with metastatic compared to localized PC and further reduced in castrate-refractory PC, indicating that ZNF132 downregulation correlates with disease progression. Reduced ZNF132 immunoreactivity was significantly associated with high Gleason score and advanced T stage in both PC patient cohorts. By univariate analysis, no/weak ZNF132 staining was a significant adverse predictor of PSA recurrence after RP (p = 0.024) and cancer-specific death following conservative treatment (p = 0.009). In multivariate models, however, ZNF132 did not add significant independent value to established prognostic factors. Finally, bisulfite sequencing revealed frequent promoter hypermethylation of ZNF132 in both PC cell lines and PC tissue samples, indicating that ZNF132 is epigenetically silenced in PC. In summary, our results show that downregulation of ZNF132 is associated with aggressive PC and furthermore identify ZNF132 as a new candidate methylation marker for PC.

Tumor-specific usage of alternative transcription start sites in colorectal cancer identified by genome-wide exon array analysis.

BACKGROUND: Approximately half of all human genes use alternative transcription start sites (TSSs) to control mRNA levels and broaden the transcriptional output in healthy tissues. Aberrant expression patterns promoting carcinogenesis, however, may arise from alternative promoter usage. RESULTS: By profiling 108 colorectal samples using exon arrays, we identified nine genes (TCF12, OSBPL1A, TRAK1, ANK3, CHEK1, UGP2, LMO7, ACSL5, and SCIN) showing tumor-specific alternative TSS usage in both adenoma and cancer samples relative to normal mucosa. Analysis of independent exon array data sets corroborated these findings. Additionally, we confirmed the observed patterns for selected mRNAs using quantitative real-time reverse-transcription PCR. Interestingly, for some of the genes, the tumor-specific TSS usage was not restricted to colorectal cancer. A comprehensive survey of the nine genes in lung, bladder, liver, prostate, gastric, and brain cancer revealed significantly altered mRNA isoform ratios for CHEK1, OSBPL1A, and TCF12 in a subset of these cancer types.To identify the mechanism responsible for the shift in alternative TSS usage, we antagonized the Wnt-signaling pathway in DLD1 and Ls174T colorectal cancer cell lines, which remarkably led to a shift in the preferred TSS for both OSBPL1A and TRAK1. This indicated a regulatory role of the Wnt pathway in selecting TSS, possibly also involving TP53 and SOX9, as their transcription binding sites were enriched in the promoters of the tumor preferred isoforms together with their mRNA levels being increased in tumor samples. Finally, to evaluate the prognostic impact of the altered TSS usage, immunohistochemistry was used to show deregulation of the total protein levels of both TCF12 and OSBPL1A, corresponding to the mRNA levels observed. Furthermore, the level of nuclear TCF12 had a significant correlation to progression free survival in a cohort of 248 stage II colorectal cancer samples. CONCLUSIONS: Alternative TSS usage in colorectal adenoma and cancer samples has been shown for nine genes, and OSBPL1A and TRAK1 were found to be regulated in vitro by Wnt signaling. TCF12 protein expression was upregulated in cancer samples and correlated with progression free survival.

Changes in phenolic compounds in garlic (Allium sativum L.) owing to the cultivar and location of growth.

The contents of total phenolic compounds, flavonoids, and phenolic acids were determined in selected garlic cultivars grown at four locations. The total phenolic content varied from 3.4 mg gallic acid equivalents (GAE)/g of dry matter (dm) to 10.8 mg GAE/g of dm with a mean value of 6.5 mg GAE/g of dm. The myricetin, quercetin, kaempferol, and apigenin flavonoids were not detected in any of the samples. Caffeic acid and ferulic acid were the major phenolic acids found with mean values of 2.9 mg/kg of dm and 2.6 mg/kg of dm, respectively. The mean contents of vanillic, p-hydroxybenzoic, and p-coumaric acids were comparable (0.4-0.8 mg/kg of dm), and the level of sinapic acid was negligible (< 0.1 mg/kg of dm). There was a significant effect of location but an insignificant effect of genotype on contents of caffeic, vanillic, p-hydroxybenzoic, and p-coumaric acids. However, genotype but not location affected the contents of total phenolics and ferulic acid. On average, the white garlic cultivars and Chinese garlic cultivars contained higher contents of total phenolics and ferulic acid than the purple garlic cultivars. However, the differences in the total phenolic content between the purple and white garlic cultivars were not significant.

Effect of genetic characteristics and environmental factors on organosulfur compounds in garlic ( Allium sativum L.) grown in Andalusia, Spain.

The content of organosulfur compounds was determined in selected garlic cultivars grown at four locations in Andalusia, Spain. The organosulfur compounds studied were three γ-glutamyl peptides, namely, γ-l-glutamyl-S-(2-propenyl)-l-cysteine (GSAC), γ-l-glutamyl-S-(trans-1-propenyl)-l-cysteine (GSPC), and γ-l-glutamyl-S-methyl-l-cysteine (GSMC), and four cysteine sulfoxides (alliin, isoalliin, methiin, and cycloalliin). There was a significant effect of the location, cultivar, and garlic ecotype on individual organosulfur compound contents. Purple-type cultivars showed on average the highest contents of GSMC, GSAC, alliin, and methiin but the lowest isoalliin content. The impact of genotype was relatively high for GSAC, whereas this factor hardly contributed to the total variability in alliin and isoalliin content. Planting date had a significant effect on the content of alliin and isoalliin. Discriminant analysis evidenced the ability of organosulfur compounds to distinguish among garlic bulbs from different locations or ecotypes with 81 or 86% accuracy, respectively.

Alternative splicing of SLC39A14 in colorectal cancer is regulated by the Wnt pathway.

Alternative splicing is a crucial step in the generation of protein diversity and its misregulation is observed in many human cancer types. By analyzing 143 colorectal samples using exon arrays, SLC39A14, a divalent cation transporter, was identified as being aberrantly spliced in tumor samples. SLC39A14 contains two mutually exclusive exons 4A and 4B and the exon 4A/4B ratio was significantly altered in adenomas (p = 3.6 × 10(-10)) and cancers (p = 9.4 × 10(-11)), independent of microsatellite stability status. The findings were validated in independent exon array data sets and by quantitative real-time reverse-transcription PCR (qRT-PCR). Aberrant Wnt signaling is a hallmark of colorectal tumorigenesis and is characterized by nuclear ß-catenin. Experimental inactivation of Wnt signaling in DLD1 and Ls174T cells by knockdown of ß-catenin or overexpression of dominant negative TCFs (TCF1 and TCF4) altered the 4A/4B ratio, indicating that SLC39A14 splicing is regulated by the Wnt pathway. An altered 4A/4B ratio was also observed in gastric and lung cancer where Wnt signaling is also known to be aberrantly activated. The splicing factor SRSF1 and its regulator, the kinase SRPK1, were found to be deregulated upon Wnt inactivation in colorectal carcinoma cells. SRPK1 was also found up-regulated in both adenoma samples (p = 1.5 × 10(-5)) and cancer samples (p = 5 × 10(-4)). In silico splicing factor binding analysis predicted SRSF1 to bind predominantly to the cancer associated exon 4B, hence, it was hypothesized that SRPK1 activates SRSF1 through phosphorylation, followed by SRSF1 binding to exon 4B and regulation of SLC39A14 splicing. Indeed, siRNA-mediated knockdown of SRPK1 and SRSF1 in DLD1 and SW480 colorectal cancer cells led to a change in the 4A/4B isoform ratio, supporting a role of these factors in the regulation of SLC39A14 splicing. In conclusion, alternative splicing of SLC39A14 was identified in colorectal tumors and found to be regulated by the Wnt pathway, most likely through regulation of SRPK1 and SRSF1.

Prediction and diagnosis of bladder cancer recurrence based on urinary content of hTERT, SENP1, PPP1CA, and MCM5 transcripts.

BACKGROUND: Identification of urinary biomarkers for detection of bladder cancer recurrence would be beneficial to minimize the frequency of cystoscopy. Our objective was to determine the usability of urine content of mRNA in the detection and prediction of bladder cancer recurrence. METHODS: We analyzed 123 prospectively cross-sectional collected urine samples from 117 patients with bladder cancer (12 incident cancers and 111 control visits). We used biopsies from cystoscopies as diagnostic criteria for recurrence, and followed the patients for a median time of 28.5 months (range 0-44 months). We measured the levels of hTERT, SENP1, PPP1CA, and MCM5 mRNA in urine by q-RT- PCR. RESULTS: We found significant differences in urinary content of hTERT (p < 0.001), SENP1 (p < 0.001), MCM5 (p < 0.001), and PPP1CA (p < 0.001) transcripts, when comparing urine samples from patients with and without tumor present in the bladder. We obtained sensitivity and specificity values for hTERT: 63/73, SENP1: 56/78, MCM5: 63/66, and PPP1CA: 69/63, respectively. Including follow-up data resulted in sensitivity and specificity values for hTERT: 62/84, SENP1:53/84, MCM5: 61/73, and PPP1CA: 65/66. Interestingly, at non-tumor visits the urinary content of especially hTERT (p = 0.0001) and MCM5 (p = 0.02) were significantly associated with subsequent tumour recurrence. Combining the markers with cytology improved the detection. The best combination was hTERT and cytology with a sensitivity of 71% and a specificity of 86% after follow-up. Further prospective validation or registration studies needs to be carried out before clinical use. CONCLUSIONS: We could use the urinary content of hTERT, SENP1, PPP1CA, and MCM5 to detect bladder cancer recurrence. All markers showed a higher sensitivity than cytology. The detection rate improved when including cytology results, but also the combination of hTERT and MCM5 increased the detection rate. Furthermore, hTERT and MCM5 levels predicted subsequent tumor recurrences.

The nuclear cofactor DOR regulates autophagy in mammalian and Drosophila cells.

The regulation of autophagy in metazoans is only partly understood, and there is a need to identify the proteins that control this process. The diabetes- and obesity-regulated gene (DOR), a recently reported nuclear cofactor of thyroid hormone receptors, is expressed abundantly in metabolically active tissues such as muscle. Here, we show that DOR shuttles between the nucleus and the cytoplasm, depending on cellular stress conditions, and re-localizes to autophagosomes on autophagy activation. We demonstrate that DOR interacts physically with autophagic proteins Golgi-associated ATPase enhancer of 16 kDa (GATE16) and microtubule-associated protein 1A/1B-light chain 3. Gain-of-function and loss-of-function studies indicate that DOR stimulates autophagosome formation and accelerates the degradation of stable proteins. CG11347, the DOR Drosophila homologue, has been predicted to interact with the Drosophila Atg8 homologues, which suggests functional conservation in autophagy. Flies lacking CG11347 show reduced autophagy in the fat body during pupal development. All together, our data indicate that DOR regulates autophagosome formation and protein degradation in mammalian and Drosophila cells.

Lysophosphatidylcholine acyltransferase 1 (LPCAT1) overexpression in human colorectal cancer.

The alteration of the choline metabolite profile is a well-established characteristic of cancer cells. In colorectal cancer (CRC), phosphatidylcholine is the most prominent phospholipid. In the present study, we report that lysophosphatidylcholine acyltransferase 1 (LPCAT1; NM_024830.3), the enzyme that converts lysophosphatidylcholine into phosphatidylcholine, was highly overexpressed in colorectal adenocarcinomas when compared to normal mucosas. Our microarray transcription profiling study showed a significant (p < 10(-8)) transcript overexpression in 168 colorectal adenocarcinomas when compared to ten normal mucosas. Immunohistochemical analysis of colon tumors with a polyclonal antibody to LPCAT1 confirmed the upregulation of the LPCAT1 protein. Overexpression of LPCAT1 in COS7 cells localized the protein to the endoplasmic reticulum and the mitochondria and increased LPCAT1 specific activity 38-fold. In cultured cells, overexpressed LPCAT1 enhanced the incorporation of [(14)C]palmitate into phosphatidylcholine. COS7 cells transfected with LPCAT1 showed no growth rate alteration, in contrast to the colon cancer cell line SW480, which significantly (p < 10(-5)) increased its growth rate by 17%. We conclude that LPCAT1 may contribute to total choline metabolite accumulation via phosphatidylcholine remodeling, thereby altering the CRC lipid profile, a characteristic of malignancy.

Translation elongation factor eEF1A binds to a novel myosin binding protein-C-like protein.

Eukaryotic translation elongation factor 1A (eEF1A) is a guanine-nucleotide binding protein, which transports aminoacylated tRNA to the ribosomal A site during protein synthesis. In a yeast two-hybrid screening of a human skeletal muscle cDNA library, a novel eEF1A binding protein, immunoglobulin-like and fibronectin type III domain containing 1 (IGFN1), was discovered, and its interaction with eEF1A was confirmed in vitro. IGFN1 is specifically expressed in skeletal muscle and presents immunoglobulin I and fibronectin III sets of domains characteristic of sarcomeric proteins. IGFN1 shows sequence and structural homology to myosin binding protein-C fast and slow-type skeletal muscle isoforms. IGFN1 is substantially upregulated during muscle denervation. We propose a model in which this increased expression of IGFN1 serves to down-regulate protein synthesis via interaction with eEF1A during denervation.

TIMP-1 expression in human colorectal cancer is associated with TGF-B1, LOXL2, INHBA1, TNF-AIP6 and TIMP-2 transcript profiles.

The balance of activity between the endogenous enzyme inhibitors known as tissue inhibitors of metalloproteinases and their targets, the matrix metalloproteinases, in the extracellular matrix is thought to play an important role in tumour cell invasion. Supporting this notion, we have shown that colorectal cancer patients have increased plasma levels of the tissue inhibitor of metalloproteinases-1 (TIMP-1), and that high plasma TIMP-1 levels are associated with short colorectal cancer patient survival. However, although TIMP-1 has been extensively studied in cancer, very little is known about how it is regulated. To further elucidate potential mechanisms of regulation of this protein, we did a number of experiments to look at associations between the transcript profile of TIMP-1 with known matrix metalloproteinases (MMPs) as well as with expression profiles of other genes differentially regulated in human colorectal cancer (CRC) and the other TIMPs 2-4, which have also been associated with the progression of colorectal cancer. Genome-wide expression profiling of 172 CRC and normal mucosa samples was used to identify transcript changes for the genes under investigation. We found that TIMP-1 was up-regulated in CRC samples compared with normal tissue, while TIMP-2 was down-regulated. Eight MMPs were up-regulated in CRC compared with normal tissue. Correlating up-regulated genes with the TIMP-1 transcript, we identified 13 that were also up-regulated in cancerous tissue. Among these were genes associated with the synthesis of extracellullar matrix, genes involved in the TGF-beta signalling pathway, and genes that are likely transcribed by the tumour cells. These insights add to the complex picture emerging about the regulation of TIMPs in colorectal cancer.