Highly-selective optical filter for NADH fluorescence detection in multiphoton microscopy.

Colorectal cancer (CRC) is a pressing global health concern, emphasizing the need for early detection tools. In this study an optical filter for precise detection of nicotinamide adenine dinucleotide (NADH) fluorescence via two-photon excitation fluorescence (TPEF) was developed. Fabricated with silicon dioxide and titanium dioxide thin films in a Fabry-Perot structure, the filter achieved a peak transmittance of about 95% at 483 nm, with a 12 nm full-width at half maximum. TPEF measurements using a tailored setup and NADH liquid phantoms underscored the filter's significance in selectively capturing NADH fluorescence while mitigating interference from other fluorophores. This work marks a substantial stride towards integrating multiphoton microscopy into conventional colonoscopy, enabling non-invasive, objective optical biopsy for colorectal tissue analysis. Further refinements of the experimental setup are imperative to advance tissue differentiation and enhance CRC diagnosis.

Synthesis of temporomandibular disorders management intervention outcomes for development of core outcome sets: A systematic review.

INTRODUCTION: The selection of appropriate outcomes in clinical trials and systematic reviews is a crucial factor in determining the results that are useful, reliable, and relevant for both patients and healthcare professionals. Clinicians and researchers have been encouraged to develop and apply core outcome sets (COS) to minimise the discrepancy between studies. AIM: This systematic review is the first phase of the COS development project for clinical trials in temporomandibular disorders (COS-TMD). It aims to identify and synthesise the outcomes used in the randomised controlled trials (RCT) that evaluated the effectiveness of interventions used in TMD management. MATERIALS AND METHODS: An electronic search was performed in several databases: MEDLINE (via PubMed), Scopus, Web of Science, Cochrane Library and EMBASE. The eligibility criteria comprised RCT that applied any intervention to treat temporomandibular joint disorders or masticatory muscle disorders. The identified outcomes were categorised according to domains of the Initiative on Methods, Measurement and Pain Assessment in Clinical Trials (IMMPACT). RESULTS: The electronic search resulted in 1606 studies. After removing duplicates and applying the eligibility criteria, 106 RCT were included. A total of 43 studies evaluated masticatory muscle disorders, 27 evaluated temporomandibular joint disorders, and 36 analysed mixed TMD. CONCLUSIONS: The evaluation showed significant variability in the types of outcomes and their measurement instruments. In addition, some domains such as physical and emotional functioning, participant ratings of global improvement and adverse events have been neglected when determining the effectiveness of treatments for TMD.

Development of clinical practice guidelines for allied health and nursing assessment and management of Duchenne muscular dystrophy.

PURPOSE: To provide evidence-based guidance specific to allied health and nursing practice for the assessment and management of individuals with Duchenne muscular dystrophy (DMD). MATERIALS AND METHODS: Thirteen key focus areas were identified in consultation with health professionals and consumer advocacy groups. A series of systematic literature reviews were conducted to identify assessment and management strategies for each key focus area. A consensus process using modified Delphi methodology, including an Australia-New Zealand expert consensus meeting, was conducted. Recommendations underwent consultative review with key groups before being finalised and prepared for dissemination. RESULTS: This clinical practice guideline (CPG) generated 19 evidence-based recommendations, 117 consensus-based recommendations and five research recommendations across the 13 focus areas to inform allied health assessment and management of individuals with DMD. CONCLUSIONS: The resulting recommendations can be used in conjunction with existing medical CPGs to improve, standardise and advocate for allied health and rehabilitation care in DMD. The process used here may be useful for the development of CPGs in other rare diseases.Implications for rehabilitationImplementation-ready evidence-based statements to guide clinical care of individuals with DMD are provided with the potential to improve participation, function in the community and quality of life.A model for developing best practice statements for other rare neurological diseases is described.Allied health and nursing health professionals should focus research efforts to generate quality evidence to support rehabilitation practice.

Profiling of antioxidant potential and phytoconstituents of Plantago coronopus / Avaliação do potencial antioxidante e perfil fitoquímico do Plantago coronopus

Abstract The halophyte species Plantago coronopus has several described ethnomedicinal uses, but few reported biological activities. This work carried out for the first time a comparative analysis of P. coronopus organs in terms of phenolic composition and antioxidant activity of organic and water extracts from roots, leaves and flowers. The leaves contents in selected nutrients, namely amino acids and minerals, are also described. Roots (ethyl acetate and methanol extracts) had the highest radical scavenging activity (RSA) towards 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, while leaves (hexane extract) had higher RSA on nitric oxide radical and iron chelating ability. High performance liquid chromatography (HPLC) analysis identified eighteen phenolics from which salicylic acid and epicatechin are here firstly described in Plantago species. Leaves had mineral levels similar to those of most vegetables, proving to be a good source for elements like calcium, sodium, iron and magnesium, and also for several of the essential amino acids justifying it use as food. Our results, especially those regarding the phenolics composition, can explain the main traditional uses given to this plantain and, altogether, emphasize the potential of P. coronopus as a source of bioactive molecules particularly useful for the prevention of oxidative stress-related diseases.

Resumo A espécie halófita Plantago coronopus tem vários usos etnomedicinais já descritos, mas em relação à bioatividade a informação é escassa. Este trabalho efetuou, pela primeira vez, uma análise comparativa dos órgãos de P. coronopus em termos de compostos fenólicos e atividade antioxidante de extratos orgânicos e aquosos provenientes das raízes, folhas e flores da planta, bem como o conteúdo de determinados nutrientes, aminoácidos e minerais, nas folhas da planta. As raízes (extratos de acetato de etila e metanol) apresentaram a maior atividade de captação para os radicais 1,1-difenil-2-picril hidrazil (DPPH) e 2,2'-azino-bis(3-etilbenzotiazolina-6-ácido sulfónico) (ABTS), enquanto as folhas (extrato de hexano) mostraram maior atividade captadora para o radical óxido nítrico bem como maior capacidade quelante do ferro. A análise por cromatografia liquida de alta eficiência (CLAE) identificou dezoito compostos fenólicos e, destes, o ácido salicílico e a epicatequina são aqui descritos pela primeira vez em espécies de Plantago. As folhas desta planta halófita mostraram ainda conter minerais em níveis semelhantes aos da maioria dos vegetais, provando ser uma boa fonte de elementos como o cálcio, sódio, ferro e magnésio, bem como de vários dos aminoácidos essenciais o que justifica seu uso na alimentação. Os resultados, particularmente aqueles relacionados à composição fenólica, podem justificar os principais usos medicinais atribuídos a esta espécie e, na sua totalidade, demonstram o potencial de P. coronopus como fonte de moléculas bioativas particularmente úteis na prevenção de doenças relacionadas com estresse oxidativo.

Profiling of antioxidant potential and phytoconstituents of Plantago coronopus.

The halophyte species Plantago coronopus has several described ethnomedicinal uses, but few reported biological activities. This work carried out for the first time a comparative analysis of P. coronopus organs in terms of phenolic composition and antioxidant activity of organic and water extracts from roots, leaves and flowers. The leaves contents in selected nutrients, namely amino acids and minerals, are also described. Roots (ethyl acetate and methanol extracts) had the highest radical scavenging activity (RSA) towards 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, while leaves (hexane extract) had higher RSA on nitric oxide radical and iron chelating ability. High performance liquid chromatography (HPLC) analysis identified eighteen phenolics from which salicylic acid and epicatechin are here firstly described in Plantago species. Leaves had mineral levels similar to those of most vegetables, proving to be a good source for elements like calcium, sodium, iron and magnesium, and also for several of the essential amino acids justifying it use as food. Our results, especially those regarding the phenolics composition, can explain the main traditional uses given to this plantain and, altogether, emphasize the potential of P. coronopus as a source of bioactive molecules particularly useful for the prevention of oxidative stress-related diseases.

Using the mini-VIDAS(®) Easy Salmonella protocol to assess contamination in transitional and coastal waters.

Classical methodologies for Salmonella detection may be too long in time to assure public safety. Presently, one of the fastest assays for Salmonella detection using the mini-VIDAS(®) system is the Easy Salmonella protocol. This assay, developed for food matrixes analysis, was here assessed for the applicability on the detection of these bacteria in transitional and saltwaters. The presence of Salmonella was detected in 4.2 % of the samples studied. In these transitional waters, the proposed protocol presented an efficiency of 79.1 %, due to a high false positive rate (20.8 %), and a false negative rate of 0 %-implying reducing analysis time, the use of enrichment broths, and making it more cost effective. Despite the multitude of samples nature, the method here described revealed to be an efficient and promising tool for transitional waters analysis.

Genetic landscape of uveal melanoma.

Uveal melanoma is genetically one of the simplest malignant tumors in adults. Initiation of these tumors is dependent of an oncogenic mutation in the GNAQ or GNA11 genes present in almost all cases. The nature of second mutational events is of major interest as it monosomy 3, gain of 8q and BAP1 inactivation are associated with unfavorable prognosis while SF3BI or EIF1AX are of good prognosis. Despite their common lineage, cutaneous and uveal melanomas are distinct diseases, implicating different oncogenic pathways and contrasting mutational landscapes. Even if uveal melanoma is a simple tumor, it is also one of the deadliest tumors in adults. There is a major clinical need for drugs targeting either the downstream pathways of Gαq and Gα11 or the biological cell functions dysregulated by BAP1 loss of function.

Benefit of adjuvant trastuzumab-based chemotherapy in T1ab node-negative HER2-overexpressing breast carcinomas: a multicenter retrospective series.

BACKGROUND: Randomized clinical trials showed the benefit of adjuvant trastuzumab-based chemotherapy (ATBC) for node-positive and/or >1 cm HER2+ breast carcinomas. No efficacy data have been published on ATBC in large series of pT1abN0 HER2+ tumors. PATIENTS AND METHODS: This retrospective study evaluated 276 cases of pT1abN0 HER2+ breast tumors in eight French cancer centers. Factors associated with prognosis and ATBC prescription were analyzed. RESULTS: A total of 129 cases (47%) were treated with ATBC (ATBC+), 19 with chemotherapy alone, 5 with trastuzumab alone, and 123 (45%) with neither trastuzumab nor chemotherapy (ATBC-). ATBC use was associated with the date of diagnosis (before or after June 2005) and with poor prognostic features. At a median follow-up of 44 months, there were 13 recurrences in the ATBC- group and 2 in the ATBC+ group. ATBC was associated with a significant survival benefit (99% 40-month disease-free survival for ATBC+ versus 93% for ATBC- cases; P = 0.018). Lack of hormone receptors (HRs) and the presence of lymphovascular invasion (LVI) were significantly associated with a poor prognosis and a greater benefit of ATBC. CONCLUSIONS: ATBC was associated with a significantly reduced risk of recurrence in pT1abN0 HER2+ tumors, and was more beneficial in HR- and/or LVI+ tumors.

Prognostic value of cardiac troponin I in patients with COPD acute exacerbation.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is frequently associated with right ventricular loading and pulmonary hypertension. We aimed to evaluate a possible association between cardiac troponin I (cTnI) levels and adverse events in hospitalised patients with acute exacerbation of COPD . METHODS: Retrospective cohort study, with analysis of admissions for acute exacerbation of COPD , with cTnI obtained in the first 48 hours of admission. A positive cTnI test was defined as 0.012 ng/ml or higher (99th percentile). Baseline and peak troponin I levels were taken as independent variables, and outcome variables included length of hospital stay, complications during hospitalisation, and in-hospital and extra-hospital mortality (evaluated 18 months post-discharge). RESULTS: Data concerned 173 patients (105 male, 68 female), with a median age of 77 years (interquartile range of 11 years). The median baseline cTnI was 0.030 ng/ml (n=173), and the median peak cTnI was 0.040 ng/ml (n=173; absolute peak value of 1.260 ng/ml). Nearly 70% of cases had a positive cTnI at admission. Both baseline and peak cTnI correlated significantly with the need for noninvasive ventilatory support. We were not able to find significant differences in in-hospital survival associated with the two troponin groups, but overall 18-month survival was significantly higher among patients with lower values of baseline and peak cTnI. CONCLUSIONS: In patients hospitalised for acute COPD exacerbations, elevated baseline and peak cTnI were associated with a greater need for noninvasive ventilatory support and were significant predictors of 18-month overall survival.

[Epidemiological link between eating habits and cancer: the example of colorectal cancer]. / Lien épidémiologique entre comportement alimentaire et cancer: exemple du cancer colorectal.

One million people worldwide are affected by colorectal cancer (CRC) every year, resulting in 500,000 deaths. Incidence rate varies widely according to areas. It has been much more common in high-income countries. This trend begins to be reversed, with a stabilization of incidence rates in high-income countries and a dramatic increase in developing regions. CRC increase seems to overlap with the increasing level of urbanization in the latter. In the eighties, Doll and Peto already reported that 80% of cancer mortality was avoidable, particularly for CRC for which environmental influence is dominating. Observational studies have reported that migrants tended to reach similar CRC incidence levels that those of natives in the host country. An estimated 60% drop of all CRC cases could be obtained provided eating habits are modified, since these seem to be the main cause of incidence variations. Over the last ten years, many epidemiological studies and meta-analyses have been conducted. However, this abundant literature gave contradictory results. Therefore, in 1997, two groups of experts: the World Cancer Research Fund (WCRF) and the American Institute of Cancer Research (AICR), considered drafting a report on the correlation between diet and cancer a necessity. This systematic literature review, updated in 2007, established that red meat, processed meat and alcohol increase the risk of CRC. Conversely fibers, milk and calcium probably protect against CRC. Considering the most important prospective studies, randomised trials and the 2nd WCRF's report, our article aims at reviewing the evidences about diet and CRC.

Weak D type 2 is the most prevalent weak D type in Portugal.

The weak D phenotype is the most common D variant, with a frequency of 0.2-1% in Caucasian individuals. There are several weak D types, with different frequencies in European countries, which may pose serologic problems and have the potential for alloimmunization. Samples from Portuguese individuals were tested for RhD by two or three distinct monoclonal and oligoclonal antisera, in direct agglutination tests. When discrepant results were observed, samples were tested with panels of monoclonal anti-D by LISS-indirect antiglobulin test. Cases that reacted weakly with IgM but positive with IgG anti-D were analysed by PCR-sequence-specific primers and real-time PCR. Ninety-nine samples were referred after being characterized as weak D. This genotype was recognized, with a preponderance of weak D type 2 (63.6%) over type 1 (16.2%) and 3 (14.1%). The high incidence of weak D type 2 in our population is in marked contrast to studies performed in other European populations and might be due to our sample selection criteria or ethnic variation. There are advantages in genotyping serologically depressed D samples to avoid the waste of D-negative RBC units and the use of immunoglobulin in pregnant women, who have no risk of alloimmunization.

Blood group antigen profile predicted by molecular biology-use of real-time polymerase chain reaction to genotype important KEL, JK,RHD, and RHCE alleles.

The most clinically important blood group systems in transfusion medicine, excluding the ABO system, are the RH, Kell, and Kidd systems. Alloantibodies to antigens of these systems may be produced following blood transfusion or during pregnancy and can result in serious hemolytic transfusion reactions and hemolytic disease of the newborn. We developed rapid and robust techniques for RHD, RHCE, KEL, and JK genotyping with the use of a real-time polymerase chain reaction instrument. Two fluorescence-based methods for the detection of amplification products were used: for KEL1/KEL2, JK1/JK2, and RHE/RHe (exon 5) we used the hybridization probes protocol; for RHC/RHc the analysis was done in sequences of exon 1 for RHC and exon 2 for RHc; and for RHD, analysis was done in sequences of intron 4, exon 7, and exon 4 pseudogene using the SYBR Green I protocol. The genotyping tests were validated with samples from 85 Caucasian Portuguese and 15 Black European blood donors. Complete phenotype-genotype correlations were obtained. The potential use of the presented methods can be predicted in clinical transfusion medicine, allowing appropriate monitoring, early intervention, and improved care. When blood group genotyping techniques are necessary, this methodology is highly competitive for a routine laboratory.

Molecular defects underlying the Kell null phenotype.

Expression of the Kell blood group system is dependent on two proteins, Kell and XK, that are linked by a single disulfide bond. Kell, a type II membrane glycoprotein, is a zinc endopeptidase, while XK, which has 10 transmembrane domains, is a putative membrane transporter. A rare phenotype termed Kell null (Ko) is characterized by the absence of Kell protein and Kell antigens from the red cell membrane and diminished amounts of XK protein. We determined the molecular basis of eight unrelated persons with Ko phenotypes by sequencing the coding and the intron-exon splice regions of KEL and, in some cases, analysis of mRNA transcripts and expression of mutants on the cell surface of transfected cells. Six subjects were homozygous: four with premature stop codons, one with a 5' splice site mutation, G to A, in intron 3, and one with an amino acid substitution (S676N) in exon 18. Two Ko persons with premature stop codons had identical mutations in exon 4 (R128Stop), another had a different mutation in exon 4 (C83Stop), and the fourth had a stop codon in exon 9 (Q348Stop). Two Ko persons were heterozygous for two mutations. One had a 5' splice site mutation (G to A) in intron 3 of one allele that caused aberrant splicing and exon skipping, and the other allele had an amino acid substitution in exon 10 (S363N). The other heterozygote had the same amino acid substitution in exon 10 (S363N) in one allele and a premature stop codon in exon 6 (R192Stop) in the other allele. The S363N and S676N mutants, expressed in 293T cells, were retained in a pre-Golgi compartment and were not transported to the cell surface, indicating that these mutations inhibit trafficking. We conclude that several different molecular defects cause the Kell null phenotype.

[Individualized monitoring of the therapy with gentamycin using pharmacokinetic methods. Which method to choose?]. / Monitorização individualizada da terapêutica com gentamicina por métodos farmacocinéticos. Que Método escolher?

Gentamicin has an excellent cost/efficacy ratio for gram negative infections treatment. Its use is often limited in clinical practice by its narrow safety margins and a high incidence of toxicity. Gentamicin related nephrotoxicity is a major adverse effect, mostly in patients with other concomitant potential risk factors. As many other Authors we have found in our Internal Medicine Service during 1992 a gentamicin related nephrotoxicity incidence of 22.5%. Various empiric methods and nomograms have shown a significant incidence of error in predicting individualized gentamicin dosage regimens. Pharmacokinetics methods have demonstrated much better results regarding efficacy and toxicity. The aim of this prospective study carried out during 1993-1994 was to individualize by pharmacokinetics methods dosage regimens of gentamicin in patients with one or more concomitant risk factors of nephrotoxicity. The purpose of pharmacokinetics dosage regimens has been to achieve trough serum concentrations of gentamicin in therapeutics range-0.5 to 2 micrograms/ml-on the first 24 to 48 hours of treatment, and the maintenance in this range during all the treatment, avoiding both toxic and under therapeutic levels. The incidence of gentamicin related nephrotoxicity has been evaluated in this population. Twenty patients were studied: 18 males and 2 females aged 59.6 years (19 to 85). All had one or more potential risk factors for nephrotoxicity-65 years or more: 13, previous renal failure: 6, other nephrotoxic drugs: 10, diuretics: 4, dehydration: 5, congestive heart failure: 5, diabetes: 3, hypertension: 3. For the first 10 patients gentamicin dosage regimens have been determined by Sawchuk-Zaske pharmacokinetics method and for the subsequent 10 patients by Bayesian method. The two subpopulations had no significant differences regarding mean age, sex and potential risk factors for nephrotoxicity. Results of Sawchuk-Zaske method: 53 trough gentamicin serum concentration were obtained; 86.8% were within the therapeutic range, 7.5% were toxic and 5.7% were under therapeutic. Results of Bayesian method: 44 determinations of gentamicin through concentrations were obtained; 86.3% within therapeutic range, 2.4% were toxic and 11.3% were under therapeutic. A great variability in pharmacokinetic patient's profile has been found and explains the great variability of individualized dosage regimens of gentamicin (30 to 320 mg/day). No patients had gentamicin related nephrotoxicity. Both pharmacokinetics methods lead to a efficient and save employment of gentamicin in patients with previous renal failure and other potential risk factors for nephrotoxicity.