Assessment of seabed litter at Concepción Seamount (Canary island) using a remotely operated towed vehicle.

The seafloor is recognised as a major sink for marine litter. However, studies conducted in this compartment addressing marine litter densities and its interactions with fauna are scarce, mainly due to sampling constraints. In this paper, we assess marine litter density, composition and interactions with marine communities and evaluate its relationship with fishing activities at the "Banco de la Concepción" seamount (Canary Islands, Spain). We took advantage of underwater video records taken with a Remotely Operated Towed Vehicle in the framework of the LIFE IP INTEMARES project. A total of 56 video transects were analysed covering about 9 km with 19 h of video recording. Transects were categorised as high, low, and null fishing effort based on the Vessel Monitoring System (VMS) positional data registered between 2009 and 2017. Litter items were recorded in 70% of the transects with a mean density of 2122 (±2464) items km-2. There were significant differences in litter densities over the three levels of fishing pressure, with a density decrease from stations of high to stations of null fishing pressure. Regarding categories, plastic was by far the most abundant category found (83.1%), mainly consisting of fishing lines, both monofilaments and entangled longlines. The study of the interactions of marine litter with fauna showed that less than 20% of the items presented an interaction with benthic organisms either by causing or not a visible impact. The sponge Asconema setubalense accounted for more than half (57.4%) of all interactions, but only 5% of all A. setubalense specimens showed physical damage.

Protocol paper: Multi-site, cluster-randomized clinical trial for optimizing functional outcomes of older cancer survivors after chemotherapy.

BACKGROUND: Cancer survivors over the age of 65 have unique needs due to the higher prevalence of functional and cognitive impairment, comorbidities, geriatric syndromes, and greater need for social support after chemotherapy. In this study, we will evaluate whether a Geriatric Evaluation and Management-Survivorship (GEMS) intervention improves functional outcomes important to older cancer survivors following chemotherapy. METHODS: A cluster-randomized trial will be conducted in approximately 30 community oncology practices affiliated with the University of Rochester Cancer Center (URCC) National Cancer Institute Community Oncology Research Program (NCORP) Research Base. Participating sites will be randomized to the GEMS intervention, which includes Advanced Practice Practitioner (APP)-directed geriatric evaluation and management (GEM), and Survivorship Health Education (SHE) that is combined with Exercise for Cancer Patients (EXCAP©®), or usual care. Cancer survivors will be recruited from community oncology practices (of participating oncology physicians and APPs) after the enrolled clinicians have consented and completed a baseline survey. We will enroll 780 cancer survivors aged 65 years and older who have completed curative-intent chemotherapy for a solid tumor malignancy within four weeks of study enrollment. Cancer survivors will be asked to choose one caregiver to also participate for a total up to 780 caregivers. The primary aim is to compare the effectiveness of GEMS for improving patient-reported physical function at six months. The secondary aim is to compare effectiveness of GEMS for improving patient-reported cognitive function at six months. Tertiary aims include comparing the effectiveness of GEMS for improving: 1) Patient-reported physical function at twelve months; 2) objectively assessed physical function at six and twelve months; and 3) patient-reported cognitive function at twelve months and objectively assessed cognitive function at six and twelve months. Exploratory health care aims include: 1) Survivor satisfaction with care, 2) APP communication with primary care physicians (PCPs), 3) completion of referral appointments, and 4) hospitalizations at six and twelve months. Exploratory caregiver aims include: 1) Caregiver distress; 2) caregiver quality of life; 3) caregiver burden; and 4) satisfaction with patient care at six and twelve months. DISCUSSION: If successful, GEMS would be an option for a standardized APP-led survivorship care intervention. TRIAL REGISTRATION: ClinicalTrials.govNCT05006482, registered on August 9, 2021.

DNA damage and aging on hematopoietic stem cells: Impact of oxidative stress in ApoE-/- mice.

The effects of aging on ROS production and DNA damage were assessed in hematopoietic stem cells (HSCs) from apolipoprotein E-deficient (ApoE-/-) mice (2-, 12- and 24-month-old), a traditional experimental model of atherogenic dyslipidemia. HSCs from aged ApoE-/- mice were associated with increased ROS levels, leading to loss quiescence, DNA damage, apoptosis and telomere shortening. The concurrence of lack of ApoE and aging result in exhaustion and senescence of HSCs accompanied by increased oxidative stress and inflammation. Therefore, our data open avenues to a better understanding of age-related changes and genetic factors, which may synergistically compromise the efficacy of aged HSC recovery and/or transplantation.

Incubation variables, performance, and morphometry of the duodenal mucosa of Japanese quails (Coturnix coturnix japonica) submitted to different incubation temperatures and thermally challenged after hatching / [Variáveis de incubação, desempenho e morfometria da mucosa duodenal de codornas japonesas (Coturnix coturnix japonica) submetidas a diferentes temperaturas de incubação e desafiadas termicamente após eclosão]

This study aimed to evaluate the effects of different temperatures on incubation variables, performance, and morphometry of the duodenal mucosa of Japanese quails (Coturnix coturnix japonica) submitted to chronic heat stress after hatching. We distributed 540 eggs in three incubators with a temperature of 37.8°C and 60% of humidity. From the 6th day of incubation until hatching, the temperatures were adjusted to (37.8°C, 38.5°C and 39.5°C). After hatching, quails were evaluated for the quality score, weighed, and distributed in a completely randomized design with three incubation temperatures (37.8, 38.5, and 39.5°C) and two ambient temperatures (stress and thermoneutral). At 10, 20, 30, and 40 days they were weighed to determine the live weight (g) and weight gain(g). To collect the duodenum and determine morphometric parameters, we euthanized four quails of each treatment. The data were analyzed, and the differences between the means determined by the Tukey test at 5%. The incubation temperature of 39.5°C provided lower hatching rate and the live weight at birth; however, from the 10th day of age, increased live weight, weight gain, and positively influenced the morphological parameters of the duodenal mucosa in situations of chronic stress.(AU)

Objetivou-se avaliar os efeitos de diferentes temperaturas de incubação sobre as variáveis de incubação, desempenho e morfometria da mucosa duodenal de codornas japonesas (Coturnix coturnix japonica) submetidas ao estresse térmico crônico por calor após eclosão. Foram distribuídos 540 ovos em três incubadoras, com temperatura de 37,8°C e umidade 60%. A partir do sexto dia de incubação até a eclosão, as temperaturas foram ajustadas para 37,8°C, 38,5°C e 39,5°C. Após a eclosão, as codornas foram avaliadas quanto ao escore de qualidade, pesadas e distribuídas em um delineamento inteiramente ao acaso, com três temperaturas de incubação (37,8ºC, 38,5ºC e 39,5°C) e duas temperaturas ambientes (estresse e termoneutro). Aos 10, 20, 30 e 40 dias, foram pesadas para determinar o peso vivo (g) e o ganho de peso(g). Quatro codornas de cada tratamento foram eutanasiadas para coleta do duodeno, para determinar os parâmetros morfométricos. Os dados foram analisados e as diferenças entre as médias foram determinadas pelo teste de Tukey a 5%. A temperatura de incubação de 39,5°C proporcionou menor taxa de eclosão e menor peso vivo ao nascer, entretanto, a partir do 10° dia de idade, essa temperatura aumentou o peso vivo, o ganho de peso e influenciou positivamente os parâmetros morfológicos da mucosa duodenal em situações de estresse crônico.(AU)

Efeito da manipulação térmica durante a incubação sobre as variáveis hematológicas, bioquímica sérica e morfometria da bolsa cloacal de codornas japonesas submetidas ao estresse crônico por calor / Effect of thermal manipulation during incubation on the hematological variables, serum biochemistry and morphometry of cloacal bursa of Japanese quails submitted to chronic heat stress

Objetivou-se avaliar os efeitos da manipulação da temperatura de incubação sobre a resposta imune de codornas desafiadas termicamente após eclosão. Para isso, foram utilizados 540 ovos, distribuídos em três incubadoras, com temperatura de 37,8°C e umidade de 60%. A partir do sexto dia de incubação até a eclosão, as temperaturas foram ajustadas em 37,8°C (padrão), 38,5°C (intermediária) e 39,5°C (alta). Após a eclosão as codornas foram pesadas e distribuídas, em delineamento inteiramente ao acaso, com três temperaturas de incubação (37,8, 38,5 e 39,5°C) e duas temperaturas de ambiente (estresse e termoneutro). Aos 10, 20, 30 e 40 dias, quatro codornas por tratamento foram eutanasiadas para coleta da bolsa cloacal, do fígado e do coração, para se determinar o peso absoluto (g), o peso relativo (%) e a área dos folículos bursais. Sangue foi coletado para realização do hemograma, do leucograma e da bioquímica sérica. Os dados foram analisados e as diferenças entre as médias foram determinadas pelo teste de Tukey a 5%. O estresse térmico por calor, a partir dos 20 dias, promove redução no peso absoluto do fígado, do coração, da bolsa cloacal e na área dos folículos bursais, além de heterofilia, linfopenia e aumento da relação heterófilo/linfócito. Em conclusão, o estresse térmico por calor após 10 dias de idade pode causar imunossupressão.(AU)

The objective of this study was to evaluate the effects of manipulation of the incubation temperature on the immune response of quails challenged thermally after hatching. For this, 540 eggs were distributed in three incubators, with temperature of 37.8°C and 60% humidity. From the 6th day of incubation to hatching the temperatures were adjusted to 37.8°C (standard), 38.5°C (intermediate) and 39.5°C (high). After hatching the quails were weighed and distributed in a completely randomized design with three incubation temperatures (37.8, 38.5 and 39.5°C) and two ambient temperatures (stress and thermoneutral). At 10, 20, 30 and 40 days four quail per treatment were euthanized to collect the cloacal burse, liver and heart to determine the absolute weight (g), relative weight (%) and area of the bursal follicles. Blood was sampled for determination of hemogram, leukogram and serum biochemistry. The data were analyzed and the differences between the means were determined by the Tukey test at 5%. Heat stress from 20 days onwards promotes a reduction in the absolute weight of the liver, heart, cloacal sac and in the area of the follicles. In addition, there was heterofilia, lymphopenia and increased heterophile/lymphocyte ratio. In conclusion, heat stress after 10 days of age can cause immunosuppression.(AU)

GHEP-ISFG collaborative exercise on mixture profiles (GHEP-MIX06). Reporting conclusions: Results and evaluation.

One of the main goals of the Spanish and Portuguese-Speaking Group of the International Society for Forensic Genetics (GHEP-ISFG) is to promote and contribute to the development and dissemination of scientific knowledge in the field of forensic genetics. Due to this fact, GHEP-ISFG holds different working commissions that are set up to develop activities in scientific aspects of general interest. One of them, the Mixture Commission of GHEP-ISFG, has organized annually, since 2009, a collaborative exercise on analysis and interpretation of autosomal short tandem repeat (STR) mixture profiles. Until now, six exercises have been organized. At the present edition (GHEP-MIX06), with 25 participant laboratories, the exercise main aim was to assess mixture profiles results by issuing a report, from the proposal of a complex mock case. One of the conclusions obtained from this exercise is the increasing tendency of participating laboratories to validate DNA mixture profiles analysis following international recommendations. However, the results have shown some differences among them regarding the edition and also the interpretation of mixture profiles. Besides, although the last revision of ISO/IEC 17025:2017 gives indications of how results should be reported, not all laboratories strictly follow their recommendations. Regarding the statistical aspect, all those laboratories that have performed statistical evaluation of the data have employed the likelihood ratio (LR) as a parameter to evaluate the statistical compatibility. However, LR values obtained show a wide range of variation. This fact could not be attributed to the software employed, since the vast majority of laboratories that performed LR calculation employed the same software (LRmixStudio). Thus, the final allelic composition of the edited mixture profile and the parameters employed in the software could explain this data dispersion. This highlights the need, for each laboratory, to define through internal validations its criteria for editing and interpreting mixtures, and to continuous train in software handling.

Correction to: Fraction of high-grade cervical intraepithelial lesions attributable to genotypes targeted by a nonavalent HPV vaccine in Galicia, Spain.

After publication of the article [1], it was brought to our attention that the author E. López-Díez is missing their second affiliation. The author would also like to indicate an affiliation to "Universidade de Vigo, Pontevedra, Spain".

Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise.

In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.

Predicting birth weight in fetuses with gastroschisis.

OBJECTIVE: To determine the accuracy of commonly utilized ultrasound formulas for estimating birth weight (BW) in fetuses with gastroschisis. STUDY DESIGN: A retrospective review was conducted of all inborn pregnancies with gastroschisis within the five institutions of the University of California Fetal Consortium (UCfC) between 2007 and 2012. Infants delivered at ⩾28 weeks who had an ultrasound within 21 days before delivery were included. Prediction of BW was evaluated for each of the five ultrasound formulas: Hadlock 1 (abdominal circumference (AC), biparietal diameter (BPD), femur length (FL) and head circumference (HC)) and Hadlock 2 (AC, BPD and FL), Shepard (AC and BPD), Honarvar (FL) and Siemer (BPD, occipitofrontal diameter (OFD), and FL) using Pearson's correlation, mean difference and percent error and Bland-Altman analysis. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for the ultrasound diagnosis of intrauterine growth restriction (IUGR) were assessed. RESULTS: We identified 191 neonates born with gastroschisis within the UCfC, with 111 neonates meeting the inclusion criteria. The mean gestational age at delivery was 36.3±1.7 weeks and the mean BW was 2448±460 g. Hadlock (1) formula was found to have the best correlation (r=0.81), the lowest mean difference (8±306 g) and the lowest mean percent error (1.4±13%). The Honarvar and Siemer formulas performed significantly worse when compared with Hadlock 1, with a 13.7% (P<0.001) and 3.9% (P=0.03) difference, respectively, between estimated and actual BW. This was supported by Bland-Altman plots. For Hadlock 1 and 2, sensitivity was 80% with a NPV of 91%. CONCLUSION: The widely used Hadlock (1) and (2) formulas provided the best estimated BW in infants with gastroschisis despite its inclusion of abdominal circumference. Furthermore, this formula performs well with diagnosis of IUGR.

A GHEP-ISFG collaborative study on the genetic variation of 38 autosomal indels for human identification in different continental populations.

A collaborative effort was carried out by the Spanish and Portuguese Speaking Working Group of the International Society for Forensic Genetics (GHEP-ISFG) to promote knowledge exchange between associate laboratories interested in the implementation of indel-based methodologies and build allele frequency databases of 38 indels for forensic applications. These databases include populations from different countries that are relevant for identification and kinship investigations undertaken by the participating laboratories. Before compiling population data, participants were asked to type the 38 indels in blind samples from annual GHEP-ISFG proficiency tests, using an amplification protocol previously described. Only laboratories that reported correct results contributed with population data to this study. A total of 5839 samples were genotyped from 45 different populations from Africa, America, East Asia, Europe and Middle East. Population differentiation analysis showed significant differences between most populations studied from Africa and America, as well as between two Asian populations from China and East Timor. Low FST values were detected among most European populations. Overall diversities and parameters of forensic efficiency were high in populations from all continents.

Fraction of high-grade cervical intraepithelial lesions attributable to genotypes targeted by a nonavalent HPV vaccine in Galicia, Spain.

BACKGROUND: Human papillomavirus (HPV) bivalent and quadrivalent vaccines have been widely implemented in worldwide organized immunization programs. A nonavalent HPV vaccine is now available in several countries. The objective was to describe the fraction of squamous non-invasive high-grade cervical intraepithelial lesions attributable to genotypes targeted by bi-quadrivalent vaccines and by nonavalent vaccine according to age and diagnosis in women living in the city of Vigo (Galicia, Spain). METHODS: Cervical scrapings (2009-2014) of women with histological diagnosis of cervical intraepithelial neoplasia grade 2 (CIN2, n = 145) and grade 3-carcinoma in situ (CIN3-CIS, n = 244) were tested with Linear Array HPV Genotyping test (Roche diagnostics, Mannheim, Germany). Hierarchical estimation of the fraction attributable to HPV 16/18 or HPV 31/33/45/52/58 detected alone or in combination was calculated. Absolute additional fraction attributable to genotypes targeted by nonavalent vaccine compared to genotypes targeted by bi-quadrivalent vaccines was calculated as the increment of attributable cases with respect to all studied cases. Age group 1, 2 and 3 included women 18 to 34, 35-44 and ≥45 years old, respectively. EPIDAT 3.1 was used. RESULTS: Fraction attributable to genotypes targeted by bi-quadrivalent vaccines was 59% CIN2 vs. 69% CIN3-CIS (p < 0.001). It was 63/51/50% of CIN2 and 78/66/45% of CIN3-CIS in age group 1, 2, 3, respectively. Fraction attributable to genotypes targeted by nonavalent vaccine was 86% CIN2 and 86% CIN3-CIS. It was 87/91/75% of CIN2 and 90/86/76% of CIN3-CIS in age group 1, 2, 3, respectively. Fraction attributable to genotypes targeted by these vaccines tended to decrease as age increased (p-trend <0.05). Globally, absolute additional attributable fraction was 16%, 26% and 29% in age group 1, 2 and 3, respectively (p < 0.005). CONCLUSIONS: Absolute additional fraction of CIN2 and CIN3-CIS attributable to genotypes targeted by nonavalent vaccine was observed in women of any age, especially in those over 35 years old.

Manejo de las vías centrales de bebés prematuros de menos de 1.500 gramos / Management of central lines in preterm babies weighing less than 1, 500 grams

Objetivo: determinar las diferencias a lo largo de los tres años en el manejo de las vías centrales de bebés prematuros menores de 1.500 gramos. Método: estudio descriptivo longitudinal retrospectivo en bebés de < 1.500 gramos ingresados en la UCI neonatal del CHUS de Santiago de Compostela, entre enero del año 2012 y diciembre de 2014. Variables de estudio: sexo, edad gestacional, peso, fecundación in vitro (FIV), gemelaridad, vía de finalización del parto, canalización o no de vía central, tipo de vía central, localización, duración, causa de retirada, tipo de germen aislado en el cultivo y exitus o no del bebé. Para el contraste de hipótesis se utilizó Anova y Chi cuadrado. Resultados: se estudiaron 120 bebés, siendo la media de edad gestacional de 29,8 semanas y el peso medio al nacimiento de 1.148 gramos. En 62 bebés (51,6%) se insertó una vía central, siendo la de primera elección la vía umbilical, con un tiempo medio de permanencia de seis días, seguido del PICC, con una permanencia media de ocho días y de localización más habitual en el miembro superior, seguida del miembro inferior y la cabeza. La causa más común de retirada de la vía central fue la electiva, debido a la finalización del tratamiento, y la contaminación más frecuente el Estafilococo epidermidis. En el periodo de estudio se encontró una leve disminución, estadísticamente significativa (p= 0,033). Conclusiones: el acceso vascular central se muestra como la principal vía de manejo del gran prematuro menor de 1.000 gramos de peso, siendo la vía umbilical la de primera elección, seguido del PICC. Aproximadamente la mitad de los bebés tuvo una vía central (AU)

Objective: to determine any differences in the management of central lines in preterm babies weighing less than 1,500 grams, during a 3-year period. Method: a descriptive longitudinal retrospective study in babies <1,500 grams admitted to the neonatal ICU at the Santiago de Compostela CHUS, between January, 2012 and December, 2014. Study variables: gender, gestational age, weight, IVF, multiple birth, type of child delivery, central venous catheterization or not, type of central line, location, duration, cause for removal, type of germ isolated in culture, and baby exitus or not. Anova and Chi Square tests were used for hypothesis contrast. Results: the study included 120 babies, with a mean gestational age of 29.8 weeks and 1,148 grams as mean weight at birth; 62 babies (51.6%) had a central line inserted, with umbilical access as first choice, for a mean period of six days, followed by peripheral catheterization, for a mean period of eight days, and the most frequent location was upper limb, followed by lower limb and head. The most frequent cause for removal of the central line was elective, due to treatment completion, and the most frequent contamination was by Staphylococcus Epidermidis. During the period of the study, a mild reduction was found, which was statistically significant (p= 0.033). Conclusions: central vascular access appears as the main pathway for management in preterm babies weighing less than 1,000 grams, with umbilical access as first choice, followed by peripheral catheterization. Approximately half of male and female babies had a central line inserted (AU)

A collaborative EDNAP exercise on SNaPshot™-based mtDNA control region typing.

A collaborative European DNA Profiling (EDNAP) Group exercise was undertaken to assess the performance of an earlier described SNaPshot™-based screening assay (denoted mini-mtSNaPshot) (Weiler et al., 2016) [1] that targets 18 single nucleotide polymorphism (SNP) positions in the mitochondrial (mt) DNA control region and allows for discrimination of major European mtDNA haplogroups. Besides the organising laboratory, 14 forensic genetics laboratories were involved in the analysis of 13 samples, which were centrally prepared and thoroughly tested prior to shipment. The samples had a variable complexity and comprised straightforward single-source samples, samples with dropout or altered peak sizing, a point heteroplasmy and two-component mixtures resulting in one to five bi-allelic calls. The overall success rate in obtaining useful results was high (97.6%) given that some of the participating laboratories had no previous experience with the typing technology and/or mtDNA analysis. The majority of the participants proceeded to haplotype inference to assess the feasibility of assigning a haplogroup and checking phylogenetic consistency when only 18 SNPs are typed. To mimic casework procedures, the participants compared the SNP typing data of all 13 samples to a set of eight mtDNA reference profiles that were described according to standard nomenclature (Parson et al., 2014) [2], and indicated whether these references matched each sample or not. Incorrect scorings were obtained for 2% of the comparisons and derived from a subset of the participants, indicating a need for training and guidelines regarding mini-mtSNaPshot data interpretation.

Analysis of uni and bi-parental markers in mixture samples: Lessons from the 22nd GHEP-ISFG Intercomparison Exercise.

Since 1992, the Spanish and Portuguese-Speaking Working Group of the ISFG (GHEP-ISFG) has been organizing annual Intercomparison Exercises (IEs) coordinated by the Quality Service at the National Institute of Toxicology and Forensic Sciences (INTCF) from Madrid, aiming to provide proficiency tests for forensic DNA laboratories. Each annual exercise comprises a Basic (recently accredited under ISO/IEC 17043: 2010) and an Advanced Level, both including a kinship and a forensic module. Here, we show the results for both autosomal and sex-chromosomal STRs, and for mitochondrial DNA (mtDNA) in two samples included in the forensic modules, namely a mixture 2:1 (v/v) saliva/blood (M4) and a mixture 4:1 (v/v) saliva/semen (M8) out of the five items provided in the 2014 GHEP-ISFG IE. Discrepancies, other than typos or nomenclature errors (over the total allele calls), represented 6.5% (M4) and 4.7% (M8) for autosomal STRs, 15.4% (M4) and 7.8% (M8) for X-STRs, and 1.2% (M4) and 0.0% (M8) for Y-STRs. Drop-out and drop-in alleles were the main cause of errors, with laboratories using different criteria regarding inclusion of minor peaks and stutter bands. Commonly used commercial kits yielded different results for a micro-variant detected at locus D12S391. In addition, the analysis of electropherograms revealed that the proportions of the contributors detected in the mixtures varied among the participants. In regards to mtDNA analysis, besides important discrepancies in reporting heteroplasmies, there was no agreement for the results of sample M4. Thus, while some laboratories documented a single control region haplotype, a few reported unexpected profiles (suggesting contamination problems). For M8, most laboratories detected only the haplotype corresponding to the saliva. Although the GHEP-ISFG has already a large experience in IEs, the present multi-centric study revealed challenges that still exist related to DNA mixtures interpretation. Overall, the results emphasize the need for further research and training actions in order to improve the analysis of mixtures among the forensic practitioners.

Gene polymorphisms and increased DNA damage in morbidly obese women.

Obesity is characterized by increased adipose tissue mass resulting from a chronic imbalance between energy intake and expenditure. Furthermore, there is a clearly defined relationship among fat mass expansion, chronic low-grade systemic inflammation and reactive oxygen species (ROS) generation; leading to ROS-related pathological events. In the past years, genome-wide association studies have generated convincing evidence associating genetic variation at multiple regions of the genome with traits that reflect obesity. Therefore, the present study aimed to evaluate the relationships among the gene polymorphisms ghrelin (GHRL-rs26802), ghrelin receptor (GHSR-rs572169), leptin (LEP-rs7799039), leptin receptor (LEPR-rs1137101) and fat mass and obesity-associated (FTO-rs9939609) and obesity. The relationships among these gene variants and the amount of DNA damage were also investigated. Three hundred Caucasian morbidly obese and 300 eutrophic (controls) women were recruited. In summary, the results demonstrated that the frequencies of the GHRL, GHSR, LEP and LEPR polymorphisms were not different between Brazilian white morbidly obese and eutrophic women. Exceptions were the AA-FTO genotype and allele A, which were significantly more frequent in obese women than in the controls (0.23% vs. 0.10%; 0.46 vs. 0.36, respectively), and the TT-FTO genotype and the T allele, which were less frequent in morbidly obese women (p<0.01). Furthermore, significant differences in the amount of genetic lesions associated with FTO variants were observed only in obese women. In conclusion, this study demonstrated that the analyzed SNPs were not closely associated with morbid obesity, suggesting they are not the major contributors to obesity. Therefore, our data indicated that these gene variants are not good biomarkers for predicting risk susceptibility for obesity, whereas ROS generated by the inflammatory status might be one of the causes of DNA damage in obese women, favoring genetically related diseases as obesity comorbidities.

Forensic ancestry analysis with two capillary electrophoresis ancestry informative marker (AIM) panels: Results of a collaborative EDNAP exercise.

There is increasing interest in forensic ancestry tests, which are part of a growing number of DNA analyses that can enhance routine profiling by obtaining additional genetic information about unidentified DNA donors. Nearly all ancestry tests use single nucleotide polymorphisms (SNPs), but these currently rely on SNaPshot single base extension chemistry that can fail to detect mixed DNA. Insertion-deletion polymorphism (Indel) tests have been developed using dye-labeled primers that allow direct capillary electrophoresis detection of PCR products (PCR-to-CE). PCR-to-CE maintains the direct relationship between input DNA and signal strength as each marker is detected with a single dye, so mixed DNA is more reliably detected. We report the results of a collaborative inter-laboratory exercise of 19 participants (15 from the EDNAP European DNA Profiling group) that assessed a 34-plex SNP test using SNaPshot and a 46-plex Indel test using PCR-to-CE. Laboratories were asked to type five samples with different ancestries and detect an additional mixed DNA sample. Statistical inference of ancestry was made by participants using the Snipper online Bayes analysis portal plus an optional PCA module that analyzes the genotype data alongside calculation of Bayes likelihood ratios. Exercise results indicated consistent genotyping performance from both tests, reaching a particularly high level of reliability for the Indel test. SNP genotyping gave 93.5% concordance (compared to the organizing laboratory's data) that rose to 97.3% excluding one laboratory with a large number of miscalled genotypes. Indel genotyping gave a higher concordance rate of 99.8% and a reduced no-call rate compared to SNP analysis. All participants detected the mixture from their Indel peak height data and successfully assigned the correct ancestry to the other samples using Snipper, with the exception of one laboratory with SNP miscalls that incorrectly assigned ancestry of two samples and did not obtain informative likelihood ratios for a third. Therefore, successful ancestry assignments were achieved by participants in 92 of 95 Snipper analyses. This exercise demonstrates that ancestry inference tests based on binary marker sets can be readily adopted by laboratories that already have well-established CE regimes in place. The Indel test proved to be easy to use and allowed all exercise participants to detect the DNA mixture as well as achieving complete and concordant profiles in nearly all cases. Lastly, two participants successfully ran parallel next-generation sequencing analyses (each using different systems) and achieved high levels of genotyping concordance using the exercise PCR primer mixes unmodified.

RNA/DNA co-analysis from human skin and contact traces--results of a sixth collaborative EDNAP exercise.

The European DNA profiling group (EDNAP) organized a sixth collaborative exercise on RNA/DNA co-analysis for body fluid/tissue identification and STR profiling. The task was to identify skin samples/contact traces using specific RNA biomarkers and test three housekeeping genes for their suitability as reference genes. Eight stains, a skin RNA dilution series and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by 22 participating laboratories using RNA extraction or RNA/DNA co-extraction methods. Two sets of previously described skin-specific markers were used: skin1 pentaplex (LCE1C, LCE1D, LCE2D, IL1F7 and CCL27) and skin2 triplex (LOR, KRT9 and CDSN) in conjunction with a housekeeping gene, HKG, triplex (B2M, UBC and UCE). The laboratories used different chemistries and instrumentation. All laboratories were able to successfully isolate and detect mRNA in contact traces (e.g., human skin, palm-, hand- and fingerprints, clothing, car interiors, computer accessories and electronic devices). The simultaneous extraction of RNA and DNA provides an opportunity for positive identification of the tissue source of origin by mRNA profiling as well as a simultaneous identification of the body fluid donor by STR profiling. The skin markers LCE1C and LOR and the housekeeping gene marker B2M were detected in the majority of contact traces. Detection of the other markers was inconsistent, possibly due to the low amounts and/or poor quality of the genetic material present in shed skin cells. The results of this and the previous collaborative RNA exercises support RNA profiling as a reliable body fluid/tissue identification method that can easily be combined with current STR typing technology.

Triple combination as adjuvant to cryotherapy in the treatment of solar lentigines: investigator-blinded, randomized clinical trial.

BACKGROUND: Post-inflammatory hyperpigmentation is a frequent concern when treating solar lentigines. OBJECTIVES: To assess the safety and efficacy of a triple combination cream with fluocinolone acetonide 0.01%, hydroquinone 4% and tretinoin 0.05% as adjuvant to cryotherapy in the treatment of solar lentigines in hands dorsum, and in the prevention of post-inflammatory hyperpigmentation after cryotherapy. METHODS: This prospective, randomized, controlled, investigator-blinded, single-centre study enrolled 50 patients. Twenty-five patients received a 2-week daily triple combination cream plus sunscreen pre-treatment and 25 received sunscreen alone. After that, cryotherapy was performed in all patients followed by a 3-week recovery period. After this period, patients received the same initial treatment and were followed up for 8 weeks. Melanin and erythema levels of a target and a control lentigo were objectively measured using a narrowband reflectance spectrophotometer. Lentigines count, colour homogeneity and global improvement were also assessed. RESULTS: The number of solar lentigines reduced in the first 2 weeks only in patients who used the triple combination 25 ± 7 vs. 22 ± 8 (P < 0.0001), and reduced at the end of the study for both groups (P < 0.0001). The melanin levels also reduced in the first 2 weeks only in patients who used the triple combination 297 ± 69 vs. 273 ± 66 (P < 0.0001) and reduced at the end of the study for both groups (P < 0.0001). Erythema and residual blisters from cryotherapy were the reported adverse reactions. CONCLUSION: Triple combination cream can be used to enhance the resolution of solar lentigines, and to significantly reduce melanin levels and lentigines count, improving treatment results. It was well-tolerated and did not increase the occurrence of neither erythema nor other side-effects after the cryotherapy.

Los conceptos que inspiran el trabajo de los psicoanalistas de pareja y familia. Estudio preliminar del análisis cuantitativo en el contenido de una encuesta / The concepts underlying the work of psychoanalyst ’s couple and family preliminary. Study of quantitative analysis on the content of a survey

En este trabajo los autores se proponen explorar qué conceptos psicoanalíticos centrales que hacen de soporte de la práctica clínica para los miembros de la Asociación Internacional de Psicoanálisis de Pareja y Familia (AIPPF). Para ello los autores han desarrollado un instrumento que permite recolectar la información (cuestionario auto administrado), que exponen brevemente. En el cuestionario se le solicita a cada miembro que mencione los conceptos de importancia en cinco terrenos: psicoanálisis individual, psicoanálisis intersubjetivo, psicoanálisis de grupo, psicoanálisis de familia, psicoanálisis de pareja, y que especifique dichos conceptos en tres niveles: el de la teoría, el de la clínica, el de la práctica y técnica. La muestra obtenida corresponde a 22 respuestas, que los autores comentan y analizan en detalle. En sus conclusiones, los autores destacan que, respecto de la terapia de pareja y familia, están presentes diferentes orientaciones teóricas, que las respuestas al cuestionario han sido meditadas, y que en ocasiones resultan originales y con un aire de modernidad. También se observa que el cambio en las configuraciones familiares en la actualidad ha dejado una marca en los modos de pensar algunos aspectos de la clínica y de los vínculos. Los autores terminan el trabajo destacando la utilidad de este trabajo para desarrollar nuevas investigaciones sobre aspectos más puntuales, entre los cuales mencionan varios que están siendo objeto de un enfoque más refinado en estudios recientes en la institución.

The author’s purpose in the present paper is to explore core psychoanalyticconcepts that lend support to clinical practice for the members of the International Association of Psychoanalysis of Couple and Family (AIPPF). To achieve this goalthe authors have developed a tool that allows to collect information (self-administered questionnaire), briefly presented. The questionnaire requests each member to sortout important concepts in the five following areas: individual psychoanalysis,intersubjective psychoanalysis, group psychoanalysis, family psychoanalysis, couple psychoanalysis, as well as to outline these concepts on three levels: theory, clinic, practice and technique. The sample obtained corresponds to 22 responses which the authors discuss andanalyze in detail. In their conclusions, the authors draw attention to the fact that different theoretical orientations are present in couple and family therapy and that theresponses to the questionnaire were pondered and on occasion are original and manifest current notions. It can also be observed that current change in family configuration shas marked the stand point concerning certain clinical aspects and links.The authors conclude by highlighting the value of this work to develop new researchon more specific aspects, among which several subjected to a more refined approachin recent studies in the institution are mentioned.