Exploring the Western Mediterranean through X-chromosome.

In this study, we investigate the forensic and population genetics properties of 21 X-chromosome markers (9 X-Alu insertions and 12 X-STRs) in a dataset composed of 716 individuals from 11 Western Mediterranean populations. The high values of combined forensic parameters indicate that this 21 X-loci panel can complement autosomal or uniparental markers in kinship analysis and complex deficient paternity testing in the populations studied. Population analyses revealed a lower differentiation between Western Mediterranean human groups for X-STRs than for X-Alu insertion polymorphisms. Moreover, X-chromosome markers suggest a sex-biased migration rate, confirming the predominance of patrilocality in this area.

Is YouTube™ a good source of information for patients to understand laparoscopic fundoplication?

INTRODUCTION: Gastro-oesophageal reflux disease affects 10-20% of the population, and laparoscopic fundoplication is one management option. As the most frequently accessed video-sharing website, YouTube has become a popular source of information for patients. This study conducted a cross-sectional analysis of the quality and demographics of patient education videos available on YouTube for laparoscopic fundoplication. METHODS: Three searches were performed on YouTube using the phrases 'laparoscopic fundoplication', 'heartburn surgery' and 'reflux operation'. The Health on The Net (HON) code, DISCERN and Journal of the American Medical Association (JAMA) systems were used to score the first 75 results from each query. Information about each video was collected, including number of views, time since posting, number of comments and the author of the video. Relationships between these variables and video quality were investigated. RESULTS: The median number of views was 3,793. The most common author category was videos produced by surgeons. Overall the quality was poor, mean HON score was 2.5/8, mean DISCERN score was 29.3/80 and mean JAMA score was 1.5/4. Surgeon-authored videos scored higher when scored using the HON and JAMA systems. Videos of longer duration scored higher using all three scoring systems. No other factors were found to be associated with video quality. CONCLUSION: The quality of information in YouTube videos on laparoscopic fundoplication is unreliable. Doctors should be aware of this and caution their patients of YouTube's limitations. Further research is needed to develop validated scoring systems for evaluating the quality of patient education videos.

Genetic sub-structure in western Mediterranean populations revealed by 12 Y-chromosome STR loci.

Haplotype and allele frequencies of 12 Y-chromosome short tandem repeat (Y-STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385 a/b, DYS437, DYS438 and DYS439) included in the Powerplex(R) Y System were determined in seven western Mediterranean populations from Valencia, Majorca, Ibiza (eastern Spain), Sicily and three Calabrian provinces (southern Italy). Amongst the 554 males included in the study, 443 different haplotypes were observed of which 372 were only observed once. The other haplotypes were shared by two to seven men. The overall haplotype diversity was 0.9988 +/- 0.0002. These Y-STRs markers showed a low capacity of discrimination (56.3%) in the Ibiza population probably due to genetic drift. Comparisons between the populations studied and other neighbouring populations showed a clear genetic sub-structure in the western Mediterranean area.

DNA extraction from soil by EZ1 advanced XL (Qiagen).

EZ1 Advanced XL (Qiagen) is an instrument designed to purify automatically nucleic acids from a wide variety of samples: different protocols for DNA extraction from forensic samples are pre-loaded into special EZ1 Advanced XL DNA Investigator Cards. The present study focuses on DNA extraction from biological samples (blood, saliva, sperm) mixed with 3 different types of soils (loam, silt, sandy), using a modified version of the Manufacturer standard protocol. In order to create several traces in the soil, we used biological samples (blood, saliva, sperm) from known donors. Quantification data, reliability and trends in STRs typing success rates using two different commercial multiplexes were evaluated. EZ1 modified DNA extraction protocol allows to recover DNA free of inhibitors and in good quantity for downstream applications.

Laparoscopic extraperitoneal repair versus open inguinal hernia repair: 20-year follow-up of a randomized controlled trial.

PURPOSE: This study compared the long-term recurrence rates of laparoscopic totally extraperitoneal (TEP) and open inguinal hernia repair in patients from a randomised trial completed in 1994. Laparoscopic inguinal hernia surgery, especially TEP repair, has gained widespread acceptance in recent years. There is still paucity of data on long-term follow-up comparing recurrence rates for open and laparoscopic techniques. This is the first study providing direct long-term comparative data about these techniques. METHODS: A randomised controlled trial was conducted between 1992 and 1994 on patients undergoing a laparoscopic TEP or an open inguinal hernia (Shouldice) repair at our institution. Of the original 104 participants, contemporary follow-up data could be obtained for 98 patients with regards to long-term recurrence. These data were collected with the help of questionnaires, telephone calls and retrieval of case records. Medical records were reviewed for all patients. Data were analysed using a Cox proportional hazards model. RESULTS: There were 7/72 (9.7%) recurrences in the open group and 9/35 (25.7%) recurrences in the laparoscopic group. This difference in recurrence rates was statistically significant (HR = 2.94; 95% CI 1.05-8.25; p = 0.041.) CONCLUSION: Laparoscopic TEP inguinal hernia repair performed in 1992-1994 had a higher recurrence rate than open Shouldice inguinal hernia repair during the same period. The original study was undertaken in the inceptive days of laparoscopic surgery and results need to be interpreted considering the technology and expertise available at that time.

Melanoma subungueal: un reto diagnóstico para los médicos / Subungual melanoma: a diagnostic challenge for doctors

El melanoma subungueal es un subgrupo del melanoma acral lentiginoso. Con frecuencia se diagnostica en etapa avanzada, dada la escasez de síntomas; de ahí que la evaluación y el tratamiento oportuno mejoran el pronóstico. Se presenta el caso de un paciente de 44 años de edad, que acude a consulta de Ortopedia con melanoniquia de tres meses de evolución. Había sido tratado en la atención primaria de salud por un hematoma secundario a un trauma directo sobre la falange distal del primer artejo del pie izquierdo. Fue seguido por consulta externa de Ortopedia y, al no experimentar una evolución adecuada, fue evaluado por la Comisión Provincial de Tumores Periféricos. Se le realizó una biopsia por punch (sacabocado), que arrojó el resultado de melanoma subungueal. Se realizó la amputación del primer artejo y del proximal del primer metatarsiano, una vez verificada la no existencia de lesiones metastásicas, con excelente resultado estético y funcional, sin mostrar recidiva en su seguimiento, a dos años de su intervención(AU)

Subungual melanoma is a subgroup of lentiginous acral melanoma. It is often diagnosed in an advanced stage by a shortage of symptoms, so timely evaluation and treatment improve prognosis. This is the case of a 44-year-old patient who goes to orthopedics with melanonichia 3 months of evolution. He had been treated for a bruise secondary to direct trauma to the distal phalanx of the first left foot ailre in primary health care. It was followed by external orthopedic consultation and by not following an adequate evolution was evaluated by the provincial commission of peripheral tumors. He was given a punch biopsy resulting in a subungual melanoma. The amputation of the first artejo and proximal first metatarsal was carried out, once verified the non-existence of metastatic lesions, with excellent aesthetic and functional result, without showing relapse in its follow-up to two years(AU)

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.

Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise.

Recently introduced rapidly mutating Y-chromosomal short tandem repeat (RM Y-STR) loci, displaying a multiple-fold higher mutation rate relative to any other Y-STRs, including those conventionally used in forensic casework, have been demonstrated to improve the resolution of male lineage differentiation and to allow male relative separation usually impossible with standard Y-STRs. However, large and geographically-detailed frequency haplotype databases are required to estimate the statistical weight of RM Y-STR haplotype matches if observed in forensic casework. With this in mind, the Italian Working Group (GEFI) of the International Society for Forensic Genetics launched a collaborative exercise aimed at generating an Italian quality controlled forensic RM Y-STR haplotype database. Overall 1509 male individuals from 13 regional populations covering northern, central and southern areas of the Italian peninsula plus Sicily were collected, including both "rural" and "urban" samples classified according to population density in the sampling area. A subset of individuals was additionally genotyped for Y-STR loci included in the Yfiler and PowerPlex Y23 (PPY23) systems (75% and 62%, respectively), allowing the comparison of RM and conventional Y-STRs. Considering the whole set of 13 RM Y-STRs, 1501 unique haplotypes were observed among the 1509 sampled Italian men with a haplotype diversity of 0.999996, largely superior to Yfiler and PPY23 with 0.999914 and 0.999950, respectively. AMOVA indicated that 99.996% of the haplotype variation was within populations, confirming that genetic-geographic structure is almost undetected by RM Y-STRs. Haplotype sharing among regional Italian populations was not observed at all with the complete set of 13 RM Y-STRs. Haplotype sharing within Italian populations was very rare (0.27% non-unique haplotypes), and lower in urban (0.22%) than rural (0.29%) areas. Additionally, 422 father-son pairs were investigated, and 20.1% of them could be discriminated by the whole set of 13 RM Y-STRs, which was very close to the theoretically expected estimate of 19.5% given the mutation rates of the markers used. Results obtained from a high-coverage Italian haplotype dataset confirm on the regional scale the exceptional ability of RM Y-STRs to resolve male lineages previously observed globally, and attest the unsurpassed value of RM Y-STRs for male-relative differentiation purposes.

Rm values of phenols. Their relationship with log P values and activity.

The experimental Rm values for a series of phenols were obtained by a reversed-phase TLC system. The extrapolation from a range of linear relationship between experimental Rm values and acetone concentration provided a set of extrapolated Rm values. This were used for studying the relationship between structure and activity in vitro and in vivo. The possibility to obtain by means of the extrapolation technique the Rm values in a standard system for serveral series of chemotherapeutic agents is pointed out.

Rm values of steroids as an expression of their lipophilic character in structure-activity studies.

The chromatographic Rm values of three series of steroids were determined by means of a reversed-phase system. The Rm values at 45% acetone in the mobile phase were shown to be correlated with the partition coefficients in an ether-water system. However, an almost equally good correlation was found when using extrapolated Rm values. The extrapolation technique could provide a standard system. The relationship between biological data and Rm values pointed out the important role of the lipophilic character in regulating the activity of steroids. In particular, the dependence of protein binding absorption and biotransformation on lipophilic character might strongly influence the availability of steroids at the site of action.

The influence of physicochemical parameters on the biliary excretion of a series of nitroimidazoles.

The relationship between physicochemical parameters and biliary excretion of nitroimidazoles was investigated. The unmetabolized form of each drug was detected in the bile by means of a UV procedure. A highly significant reversed parabolic relationship was shown between the Rm values and the biliary excretion of the test compounds. In other words, the compounds closer to the optimal Rm value are excreted less than those characterized by higher or lower Rm values. Since the Rm values seem to account for both the lipophilic and polar character of nitroimidazoles, the reversed parabola could be due to plasma protein binding and/or some protein binding within the hepatocyte. In fact, both the lipophilic and polar character seem to play an important role in protein binding of chemicals.

Relationship between lipophilic character and urinary excretion of nitroimidazoles and nitrothiazoles in rats.

Many processes are involved in the renal excretion of drugs, but very little is known about their quantitative structure-activity relationship. The relationship between urinary excretion and lipophilic character of a series of nitroimidazoles and nitrothiazoles was studied. The unmetabolized forms of the drugs were detected in the urine by means of UV and HPLC procedures. The urinary excretion of unmetabolized forms is parabolically related with the log P, as an expression of lipophilic character of molecules.

Rm values and structure-activity relationship of benzodiazepines.

Quantitative structure-activity relationships (QSAR) have been formulated for the activities of a series of benzodiazepines in rats. The lipophilic character of molecules was expressed by means of the chromatographic Rm values which were very well correlated with experimental or calculated log P values. The ideal lipophilic character for activity of benzodiazepines in the exploratory behavior test is not far from that of compounds acting in the central nervous system as unspecific depressant agents. The results of both the conflict and exploratory behavior studies might support the hypothesis of different sites of action for the antianxiety and sedative effects of benzodiazepines.

About an unusual case: GC-MS detection of lidocaine.

A man suspected to be a drug user was found dead; toxicological analysis of his biological samples showed the presence of lidocaine in all samples analysed, while cocaine was found only in the blood, urine and hair samples. We found that the cause of death was determined by ventricular arrhythmia associated to lidocaine/cocaine overdosage.

DNA analysis from mixed biological materials.

At the crime scene a mixture of biological samples from more than one individual often happens and since generally different contributors may give in the mixture a different contribution of biological material, in this study we have evaluated this effect on STRs analysis.

Detection of STRs from body fluid collected on IsoCode paper-based devices.

Isocode card and IsoCode ID Sample Registration Matrix (Schleicher and Schuell BioScience Inc.) are different paper devices chemically treated that keep sample DNA safe from degradation, allowing for the indefinite storage of DNA at ambient temperatures. In this study, blood was collected on Isocode card while saliva, sperm were collected on IsoCode ID. We also tried to verify the effect of collecting urine on Isocode ID. All samples (belonging to the same male donor) were analyzed after 1 day, 1-5 months since their application on the paper.

Anonymous letters? DNA and fingerprints technologies combined to solve a case.

Two brothers, living in two different cities, received two different anonymous letters. We performed latent prints development and DNA research on the letters and also on a glass used by a cousin suspected to be the letters' sender.

Simultaneous estimation of a Y-specific fragment, an X-specific fragment and sex determination of forensic studies in real-time PCR.

The human sex test in forensic multiplexes is based on the amelogenin gene on both the X and Y chromosomes commonly used in sex genotyping. In this work sex determination across the quantification of nuclear DNA was achieved by real-time PCR of a segment of the X-Y homologous amelogenin (AMG) gene that allowed the simultaneous estimation of a Y- and an X-specific fragment. The AMG real-time PCR design has been used to quantify a set of forensic casework samples.

Rapid and efficacious real-time quantitative PCR assay for quantitation of human DNA in forensic samples.

A reliable quantification of human DNA is necessary in the process of routine forensic human identification. When DNA is degraded, contaminated or associated with inhibitors, is important to accurately estimate the concentration of extracted DNA prior to perform an analysis based on nuclear markers amplified. In this work, a new approaches to DNA quantification employees the use of a real-time fluorescence probe system. The real-time PCR assay is highly sensitive, specific, rapid, cost-effective and flexible assay for analysis of forensic casework samples, can perform with DNA of poor quality and detect specifically amplifiable DNA rather than total DNA for STR analysis.

A case of Wilson's syndrome.

A young girl (16 years old), with a grave type of hepatitis and advanced cirrhosis of liver, died 4 days after her hospitalisation. To verify the real causes of death, it was required the autopsy. After toxicological and histo-pathological analysis it was suspected a case of Wilson's syndrome.