Unveiling the mechanisms underlying photothermal efficiency of gold shell-isolated nanoparticles (AuSHINs) on ductal mammary carcinoma cells (BT-474).

Gold nanoparticles are valuable photothermal agents owing to their efficient photothermal conversion, photobleaching resistance, and potential surface functionalization. Herein, we combined bioinspired membranes with in vitro assays to elicit the molecular mechanisms of gold shell-isolated nanoparticles (AuSHINs) on ductal mammary carcinoma cells (BT-474). Langmuir and Langmuir-Schaefer (LS) films were handled to build biomembranes from BT-474 lipid extract. AuSHINs incorporation led to surface pressure-area (π-A) isotherms expansion, increasing membrane flexibility. Fourier-transform infrared spectroscopy (FTIR) of LS multilayers revealed electrostatic AuSHINs interaction with head portions of BT-474 lipid extract, causing lipid chain disorganization. Limited AuSHINs insertion into monolayer contributed to hydroperoxidation of the unsaturated lipids upon irradiation, consistently with the surface area increments of ca. 2.0%. In fact, membrane disruption of irradiated BT-474 cells containing AuSHINs was confirmed by confocal microscopy and LDH leakage, with greater damage at 2.2 × 1013 AuSHINs/mL. Furthermore, the decrease in nuclei dimensions indicates cell death through photoinduced damage.

Candidate biomarkers of physical frailty in heart failure: an exploratory cross-sectional study.

AIMS: Physical frailty is highly prevalent and predictive of worse outcomes in heart failure (HF). Candidate biomarker analysis may help in understanding the mechanisms underlying physical frailty in HF. We aimed to identify candidate biomarkers associated with physical frailty in HF using a multimarker strategy of distinct pathophysiological processes. METHODS AND RESULTS: We collected data and plasma samples from 113 adults with New York Heart Association Functional Class I-IV HF. Physical frailty was measured with the Frailty Phenotype Criteria. Plasma biomarkers included: N-terminal pro-B-type natriuretic peptide, norepinephrine, dihydroxyphenylglycol, soluble tumour necrosis factor alpha receptor-1, adiponectin, insulin, glucose, insulin-like growth factor-1 (IGF-1), and myostatin. Comparative statistics and multivariate linear regression were used to test group differences and associations. The average age was 63.5 ± 15.7 years, half were women (48%), and most had a non-ischaemic aetiology of HF (73%). Physical frailty was identified in 42% and associated with female sex, higher body mass index and percent body fat, more comorbidities, and HF with preserved ejection fraction. Adjusting for Seattle HF Model projected survival score, comorbidities, body composition, and sex, physical frailty was associated with significantly lower plasma adiponectin [ß ± standard error (SE) -0.28 ± 0.14, P = 0.047], IGF-1 (ß ± SE -0.21 ± 0.10, P = 0.032), and myostatin (ß ± SE -0.22 ± 0.09, P = 0.011). In sex-stratified analyses, IGF-1 and myostatin were significantly associated with physical frailty in men but not women. CONCLUSION: We identified biomarkers involved in adipose tissue and skeletal muscle development, maintenance, and function that were associated with physical frailty in HF.

Plasma levels of efavirenz and frequency of the CYP2B6 516G>T polymorphism in people living with HIV-1 in Mexico.

Efavirenz (EFV) is a widely used antiretroviral, due to its safety, efficacy, and low cost. However, plasma concentrations have been related with an increased risk of virological failure and the appearance of serious adverse reactions. EFV is metabolized by Cytochrome P450, the main isoenzyme involved is CYP2B6 and the most relevant genetic polymorphisms found in several populations has been the CYP2B6 516G> T. The aim of this study was to identify the frequency of the CYP2B6 516G>T polymorphism and its effect on the plasma concentration of efavirenz (EFV) in a group of people living with HIV (PLWH) and undergoing EFV treatment in Morelos, Mexico. Ninety-six PLWH undergoing EFV treatment, at a daily dose of 600 mg orally in combination with other antiretrovirals (ARVs), were included in this study. The CYP2B6 516G>T polymorphism was detected using PCR-RFLP. The plasma concentrations of EFV were evaluated by high-resolution liquid chromatography coupled to a mass-mass detector, using a protein precipitation method. The median plasma EFV concentration was 4.6 µg/mL (IQR = 4.64) and 64.6% of the subjects had concentrations above the therapeutic range. The CYP2B6 516G>T genotype findings were as follows: 46.9% of the population presented the wild-type genotype (GG), while 45.8 % and 7.3 % showed the heterozygote (GT) and the polymorphic homozygote (TT) genotype, respectively. The homozygote G had the lowest plasma concentrations of EFV (median = 4.1 µg/mL and IQR = 1.7 µg/mL), followed by those with the GT genotype (median = 5.1 µg/mL and IQR = 3.0 µg/mL). Participants with the homozygous T genotype had the highest EFV concentrations (median = 9.7 µg/mL and IQR = 5.8 µg/mL). In conclusion, the CYP2B6 516G>T polymorphism was associated with plasma levels of EFV in PLWH undergoing ARV treatment. EFV plasma concentrations at 600mg doses were outside the therapeutic range in most subjects.

Riesgo de contagio por COVID-19 en función del tipo de contacto y de la renta familiar / Risk of contagion by COVID-19 depending on the type of contact and family income

Introducción: tras una segunda oleada de la pandemia COVID-19 en Zaragoza, se estudian los contactos estrechos con pacientes con COVID-19, con la finalidad de valorar el riesgo de infección tras exposición en función del paciente índice, adulto o niño, y la renta familiar. Material y métodos: se realiza un estudio descriptivo de todos los pacientes pediátricos que han tenido contacto con pacientes con COVID-19 en el centro de salud de Delicias Sur de Zaragoza entre los meses de julio y agosto de 2020 y se valora la aparición de síntomas, consultas en servicio de urgencias, realización de pruebas complementarias, contacto estrecho con adulto o con niño y la renta familiar. Resultados: un total de 292 pacientes fueron contacto estrecho con pacientes con COVID-19, de los cuales 128 fueron positivos para la PCR de SARS-CoV-2. Al analizar el tipo de contacto, se encontró que un 10,94% había mantenido contacto estrecho con un niño y un 89,06% con un adulto. El riesgo de contagio tras sufrir contacto con un niño positivo fue del 34,15%, mientras que, tras contacto con un adulto positivo, el riesgo fue del 45,78%. Asimismo, el riesgo de contagio entre las rentas inferiores a 18 000 € fue de 47,9 frente al 27,6% de aquellos pacientes con rentas superiores a 18 000 €. Conclusiones: el riesgo de contagio de COVID-19 es mayor cuando el contacto estrecho es con un adulto y cuando el nivel de renta familiar es inferior (AU)

Purpose: during the second wave of the COVID-19 pandemic in Zaragoza, we studied the paediatric contacts of COVID-19-positive patients to estimate the risk of infection after exposure to a positive child or adult and the risk of infection based on household income.Methods: we conducted a descriptive study of all paediatric patients that were close contacts of individuals with COVID-19 in the Delicias Sur Primary Care Centre (Zaragoza, Spain) between July and August 2020. We also analysed the most frequent symptoms, visits to the emergency department, diagnostic tests, contact with a child versus an adult with COVID-19 and household income.Results: a total of 292 patients had had close contact with individuals with COVID-19; 218 of them had positive SARS-CoV-2 PCR test results. When we analysed the close contacts, we found that 10.94% of patients that tested positive had been in close contact with a child with COVID-19 and 89.06% with an adult with COVID-19. The estimated risk of infection after exposure was 29.8% in the case of close contact with a child with COVID-19 compared to 46.53% when it came to close contact with an adult case. The risk of infection was higher in patients with an annual household income of less than €18000 (47.9%) compared to patients with a higher annual household income (27.6%).Conclusion: the risk of SARS-CoV-2 infection was higher in patients that had close contact with adult cases and with lower household incomes. (AU)

Background and Design of the Biological and Physiological Mechanisms of Symptom Clusters in Heart Failure (BIOMES-HF) Study.

BACKGROUND: Symptoms, which often cluster together, are a significant problem in heart failure (HF). There is considerable heterogeneity in symptom burden, particularly in the vulnerable transition period after a hospitalization for HF, and the biological underpinnings of symptoms during transitions are unclear. The purpose of this article is to describe the background and design of a study that addresses these knowledge gaps, entitled Biological and Physiological Mechanisms of Symptom Clusters in Heart Failure (BIOMES-HF). METHODS AND RESULTS: BIOMES-HF is a prospective gender- and age-balanced longitudinal study of 240 adults during the 6-month transition period after a HF hospitalization. The aims are to (1) identify clusters of change in physical symptoms, (2) quantify longitudinal associations between biomarkers and physical symptoms, and (3) quantify longitudinal associations between physical frailty and physical symptoms among adults with HF. We will measure multiple symptoms, biomarkers, and physical frailty at discharge and then at 1 week and 1, 3, and 6 months after hospitalization. We will use growth mixture modeling and longitudinal mediation modeling to examine changes in symptoms, biomarkers, and physical frailty after HF hospitalization and associations therein. CONCLUSIONS: This innovative study will advance HF symptom science by using a multibiomarker panel and the physical frailty phenotype to capture the multifaceted nature of HF. Using advanced quantitative modeling, we will characterize heterogeneity and identify potential mechanisms of symptoms in HF. As a result, this research will pinpoint amenable targets for intervention to provide better, individualized treatment to improve symptom burden in HF. LAY SUMMARY: Adults with heart failure may have significant symptom burden. This study is designed to shed light on our understanding of the role of biological and physiological mechanisms in explaining heart failure symptoms, particularly groups of co-occurring symptoms, over time. We explore how symptoms, biomarkers, and physical frailty change after a heart failure hospitalization. The knowledge generated from this study will be used to guide the management and self-care for adults with heart failure.

α-Glucosidase inhibitory phenolics from Echeveria subrigida (B. L. Rob & Seaton) leaves.

Echeveria subrigida is native to Mexico and its methanol extract (ME) shows relevant biological activities for human health, including the α-glucosidase inhibitory (αGI) activity that suggests its antidiabetic potential. Fractionation of the ME based on the αGI activity (IC50 in µg/mL) showed that quercetin-3-O-ß-glucoside (131.1), isorhamnetin-3-O-ß-glucoside (166.4), and dimers to heptamers proanthocyanidins (9.6) were among the main responsible of αGI activity in the ME. The purified compounds showed better activity than acarbose (IC50 = 4426 µg/mL).

Characterizing Sex Differences in Physical Frailty Phenotypes in Heart Failure.

BACKGROUND: Although women with heart failure (HF) are potentially more likely to be physically frail compared with men with HF, the underlying contributors to this sex difference are poorly understood. The purpose of this study was to characterize sex differences in physical frailty phenotypes in HF. METHODS: We prospectively enrolled adults with class I-IV HF. Physical frailty was measured with the frailty phenotype criteria. Symptoms of dyspnea, sleep-related impairment, pain interference, depression, and anxiety were assessed. Body composition was measured using dual-energy x-ray absorptiometry. Simple comparative statistics and stepwise regression modeling were used. RESULTS: The average age of the sample (n=115) was 63.6±15.7 years, 49% were women, and 73% had nonischemic cause. Forty-three percent of the sample was physically frail. Women had a 4.6 times greater odds of being physically frail compared with men, adjusting for covariates (odds ratio=4.63 [95% CI, 1.81-11.84], P=0.001). Both physically frail men and women were characterized by more type 2 diabetes, higher comorbidity burden, and worse dyspnea symptoms. Physically frail women had significantly worse symptoms compared with non-physically frail women but no difference in body composition characteristics. Physically frail men had significantly lower appendicular muscle mass, higher percent fat, lower hemoglobin, and more depressive symptoms compared with non-physically frail men. CONCLUSIONS: Women are significantly more likely to be physically frail compared with men in HF. Physical frailty in both women and men is characterized by comorbidities and worse symptoms; physical frailty in men is characterized by worse physiological characteristics.

Association of human beta-defensin 1 gene polymorphisms with nonsegmental vitiligo.

BACKGROUND: Vitiligo is a pigmentation disorder of autoimmune aetiology. Polymorphisms in beta-defensin genes have been linked to a predisposition to some autoimmune disorders. AIM: To evaluate the role of polymorphisms in DEFB1, the gene encoding for human beta-defensin (HBD)-1 and its 5' untranslated region in nonsegmental vitiligo. METHODS: In total, 354 participants [171 patients with non-segmental vitiligo and 183 age and sex-matched healthy controls (HCs)], were genotyped by the PCR-restriction fragment length polymorphism (RFLP) method. For 80 of these individuals (40 patients and -40 HCs) serum HBD-1 was also measured by ELISA. RESULTS: The -44 G allele, CG genotype and GGG haplotype increased the risk for vitiligo (P < 0.02 in all cases), whereas the -20 AA genotype seems to be protective (P = 0.04). Serum HBD-1 levels were lower in patients with vitiligo than in HCs (P < 0.01), as well as in patients with active vitiligo compared with those with stable vitiligo and with HCs (P < 0.05 in both cases), CONCLUSION: Our results suggest that HBD-1 and its gene polymorphisms may modulate vitiligo susceptibility and/or disease activity. This is the first report, to our knowledge, of the association of serum HBD-1 levels and DEFB1 gene polymorphisms with vitiligo.

[Biomechanical analysis through numerical simulation of rupture of the tibial posterior tendon in valgus flat foot: a cadeveric study]. / Análisis biomecánico a través de simulación numérica de la ruptura del tendón del tibial posterior en el pie plano valgo: estudio en cadáver.

The insufficiency of the posterior tibial tendon is the most common acquired cause of pain related to valgus flatfoot deformity in adults. The acquired flatfoot adult is a very painful symptomatic deformity resulting from a gradual stretching (attenuation) of the posterior tibial tendon and ligaments that support the arch of the foot. The progressive pain acquired flatfoot adult affects four times more women than men. Some factors that contribute to increased risk of acquired flatfoot in adults, are diabetes, hypertension and obesity. It is thought that the combination of the following events is the cause of acquired flatfoot adult. Johnson Strom classification modified by Mayerson evaluates in 4 stages. This study was divided into 3 stages: Stage 1: Dissection and three-dimensional analysis of the tendon, Step 2: Application of tools bioengineering to determine the causes of rupture of the tibial tendon: Stage 3: Evaluation of 24 patients with flatfoot disease valgus for describe the deformity.

La insuficiencia del tendón del tibial posterior es la causa adquirida de dolor más común relacionada con la deformidad de pie plano valgo en adultos. El pie plano adquirido por el adulto es una deformidad sintomática muy dolorosa que resulta de un estiramiento gradual (atenuación) del tendón del tibial posterior, así como de los ligamentos que sostienen el arco de la planta del pie. El dolor progresivo en el pie plano adquirido por el adulto afecta cuatro veces más a las mujeres que a los hombres. Algunos factores que contribuyen a incrementar el riesgo del pie plano adquirido en el adulto son la diabetes, la hipertensión y la obesidad. Se piensa que la combinación de los eventos siguientes es la causa del pie plano adquirido por el adulto. La clasificación de Johnson Strom modificada por Mayerson lo evalúa en cuatro estadios. Este estudio se dividió en tres etapas: etapa 1: disección y análisis tridimensional del tendón, etapa 2: aplicación de herramientas de bioingeniería para determinar las causas de ruptura del tendón del tibial posterior y etapa 3: evaluación de 24 pacientes con enfermedad de pie plano valgo para describir la deformidad.

Immunoassay quantification using surface-enhanced fluorescence (SEF) tags.

Fluorescence-based immonoassays are widely used in several areas, ranging from basic biomedical research to disease diagnostics. A variety of new probes have been developed recently to address some limitations in typical assays performed with organic dyes. Ideally, new fluorescence tags that allow quantification with a low limit of detection are highly desired. In this work, the surface-enhanced fluorescence (SEF) phenomenon was explored in the development of tags for Immunoglobulin-M (IgM) detection. Shell-isolated gold nanoparticles (Au-SHINs) with 100 nm core size and a 10 nm silica shell were synthesized. These particles contain an outermost thin fluorescent layer of nile blue (NB) that was further coated by another 5 nm of silica (SEF tags). The outer silica shell was then functionalized with antibodies to allow the detection of IgM as in typical immunological sandwich assays. IgM concentrations down to the 10 ng mL-1 mark were successfully detected. A linear dependence between the average fluorescence intensity and the IgM concentration was found.

Palliative care and decision making in advanced heart failure.

OPINION STATEMENT: The diagnosis of advanced heart failure (HF) is established in patients for whom symptoms are refractory to guideline-directed therapies. Palliative care (PC) is based on symptom management and support of the patient and family, making its integration into the care of those with advanced HF essential. Comorbidities including frailty, cognitive dysfunction, and depression are often under-recognized in patients with advanced HF and may correlate with outcomes. Decisions should be based on the patient's values, goals agreed upon by the clinician with the patient, and what is medically reasonable. Palliative Care should be integrated to help with both palliation of symptoms and support for families and patients.

Physical and sensory characterizations of oral coatings of oil/water emulsions.

The physical and sensory properties of oil coatings on the tongue formed by five oil/water emulsions varying in oil content were investigated. A total of 20 subjects processed orally each emulsion for 30 s in triplicate. In vivo fluorescence measurements at the front and back of the anterior tongue were made to quantify the oil fraction deposited at different time points. Calibration lines relating fluorescence intensity to oil fraction were determined using pig tongues at 37.5 °C to mimic oral conditions. The oil fraction on the tongue increased linearly with an increasing oil content of the emulsions. The oil fraction deposited at the back of the anterior tongue was 1.5-2.0× larger than at the front. The intensity of sensory attributes describing after-feel perception was related to the oil fraction by Weber-Fechner's law. This study uses in vivo fluorescence to study food behavior in the mouth and unravel new insights in after-feel perception of emulsions.

Rectal hyposensitivity for non-noxious stimuli, postprandial hypersensitivity and its correlation with symptoms in complete spinal cord injury with neurogenic bowel dysfunction.

STUDY DESIGN: Prospective clinical study. OBJECTIVES: To assess fasting and postprandial (PP) perception of rectal distension and its correlation with symptoms in patients with spinal cord injury (SCI) and neurogenic bowel dysfunction compared to ten healthy subjects (HS). SETTING: Experimental Medicine and Motility Unit, Mexico General Hospital and National Institute of Rehabilitation. METHODS: Twenty patients with complete SCI at cervical, thoracic and lumbar levels [American Spinal Injury Association (ASIA) A] were studied. Rectal sensitivity was evaluated with a barostat. RESULTS: In SCI patients, while lower the rectal tone more time was used for defecate (R=0.50, P=0.048) and more PP episodes of fecal incontinence occur (R=0.54, P=0.030). The thresholds for non-noxious stimuli of first (23.6 mmHg, CI 19.5-27.7) vs 14.0 (CI 10.9-17.1), P=0.004; gas (27.9 mmHg, CI 19.9-35.8) vs 17.9 mmHg (CI 14.25-21.69), P=0.02 and urge-to-defecate sensation (33.2 mmHg, CI 27.5-38.8) vs 22.4 mmHg (CI 17.9-26.9), P=0.01 were reported by SCI patients at higher pressure than HS, respectively. SCI patients reported PP pain sensation at a lower pressure than controls (27.8 mmHg, CI 21.5-34.2 vs 36.5 mmHg, CI 31.8-41.2), P=0.04. CONCLUSION: SCI patients preserve rectal sensation, present rectal hyposensitivity for non-noxious stimuli and PP hypersensitivity. Lower rectal tone was related to the time used for defecate and with fecal incontinence. The results suggest that an intact neural transmission between the spinal cord and higher centres is indispensable for noxious stimulus, but not for non-noxious stimuli. Also, barostat sensitivity studies can complement ASIA criteria to verify a complete injury.

An assessment of the use of drug and non-drug interventions in the treatment of Ichthyophthirius multifiliis Fouquet, 1876, a protozoan parasite of freshwater fish.

Infection by the ciliate protozoan Ichthyophthirius multifiliis Fouquet, 1876 causes significant economic losses in freshwater aquaculture worldwide. Following the ban on the use of malachite green for treating food fish, there has been extensive research aimed at identifying suitable replacements. In this paper we critically assess drug and non-drug interventions, which have been tested for use or have been employed against this parasite and evaluate possibilities for their application in farm systems. Current treatments include the administration of formaldehyde, sodium chloride (salt), copper sulphate and potassium permanganate. However, purportedly more environmentally friendly drugs such as humic acid, potassium ferrate (VI), bronopol and the peracetic acid-based products have recently been tested and represent promising alternatives. Further investigation, is required to optimize the treatments and to establish precise protocols in order to minimize the quantity of drug employed whilst ensuring the most efficacious performance. At the same time, there needs to be a greater emphasis placed on the non-drug aspects of management strategies, including the use of non-chemical interventions focusing on the removal of free-swimming stages and tomocysts of I. multifiliis from farm culture systems. Use of such strategies provides the hope of more environmentally friendly alternatives for the control of I. multifiliis infections.

In vitro treatments for the theront stage of the ciliate protozoan Cryptocaryon irritans.

The ciliate protozoan Cryptocaryon irritans Brown, 1951, the 'marine white spot', causes one of the most important parasitic fish diseases, with extensive losses every year in mariculture and in the ornamental fish industry. In the present study, we explore the in vitro use of 8 different compounds against the theront (infective) stage of C. irritans; these compounds include extracts of natural products (epigallocatechin gallate (EGCG), L-DOPA, papain), peracetic acid-based compounds (Proxitane 5:23 and 15% peracetic acid, PAA), quinine-based compounds (quinacrine hydrochloride and chloroquine diphosphate) and hydrogen peroxide. All of these compounds had an effect on theront survival; however, only EGCG caused significant theront mortality when applied in doses > or =50 mg l(-1) and over a period of 3 h; papain caused a maximum theront mortality of <50%. We discuss the type of application and potential utility of the compounds tested as part of a management control strategy for C. irritans infections in marine aquaculture and the ornamental fish industry.

Serodiagnostic reliability of single-step enriched low-molecular weight proteins of Taenia solium metacestode of American and Asian isolates.

The low-molecular weight proteins (LMWPs) of Taenia solium metacestode (TsM) constituted pertinent serodiagnostic antigens for cysticercosis. We established a novel single-step purification of the LMWPs from TsM cyst fluid (CF). When the CF was precipitated with trichloroacetic acid/acetone mixture at the final concentrations of 5 and 50%, most LMWPs (ranging 7-38kDa) remained in the supernatant fraction. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses revealed that the LMWPs consisted mainly with the previously reported subunit proteins of the 120- and 150-kDa antigen complexes. Serum samples from neurocysticercosis (NC) and other helminthic infections, as well as those from healthy controls were tested by both immunoblotting and micro-ELISA. In 135 cases of active stage NC patients, 132 cases (97.7%) showed positive reactions. Serum samples from other helminthic diseases (n=125) and healthy controls (n=100) exhibited no positive reactions except for cystic echinococcosis, of which 12% (3/25 cases) exhibited low levels of cross-reactivity. The LMWPs from different geographical regions (Korea and Mexico) showed diagnostic sensitivity and specificity of 97.7% and 98.7% against active stage NC. Our single-step separation method for the LMWPs provided excellent performance with easy applicability and high reproducibility, which has a great benefit for preparation of potent antigen in endemic areas.

Trichuriasis and low-iron status in schoolchildren from Northwest Mexico.

BACKGROUND/OBJECTIVES: The aim of this study was to investigate the association between trichuriasis and iron status in rural schoolchildren from Northwest Mexico. SUBJECTS/METHODS: A total of 73 schoolchildren (37 boys, 36 girls) between the ages of 6 and 10 years were voluntarily recruited from the public primary school of the rural community of El Higueral in Culiacan, Sinaloa (Northwest Mexico) for a cross-sectional study with a longitudinal follow-up of 5 weeks. Data were collected on socioeconomic status, anthropometry, haematological and biochemical indices of iron status, daily iron intake, and prevalence and intensity of trichuriasis. Multiple regression models, independent t-test and paired t-test were used to analyse the association between trichuriasis and iron status in cross-sectional and longitudinal samples, respectively. Adjusted models were tested for linear regression assumptions using residual plots. RESULTS: The mean age of the Trichuris-free and Trichuris-infected groups was 7.7±1.3 and 7.7±1.5 years respectively (P=0.92). The height for age was significantly higher in the Trichuris-free group than the Trichuris-infected group (P=0.02). No differences were found in the socioeconomic variables between the two groups. At baseline, significantly higher concentrations of haemoglobin, haematocrit, blood cell count (RBC) and serum iron were measured in the Trichuris-free group compared with the Trichuris-infected children (P<0.05). An association was found between trichuriasis and haemoglobin adjusted for socioeconomic variables, age and sex. Haemoglobin, RBC and serum ferritin concentrations were significantly increased in the infected children 5 weeks after treatment (P<0.05). CONCLUSIONS: Trichuriasis could be a risk factor for low-iron status in the schoolchildren of Northwest Mexico.

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea).

Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss' Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a "G. salaris-like" (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select "G. salaris-like" specimens, which are subsequently identified by molecular-based techniques.