Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer.
Zygote. 2016 Dec 08;:1-13
Authors: Arias ME, Sánchez-Villalba E, Delgado A, Felmer R
Sperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.
PMID: 27928970 [PubMed - as supplied by publisher]
A Bayesian Approach to Age-at-Death Estimation from Osteoarthritis of the Shoulder in Modern North Americans.
J Forensic Sci. 2016 Dec 08;:
Authors: Brennaman AL, Love KR, Bethard JD, Pokines JT
Osteoarthritis (OA) is a marker of degeneration within the skeleton, frequently associated with age. This study quantifies the correlation between OA and age-at-death and investigates the utility of shoulder OA as a forensic age indicator using a modern North American sample of 206 individuals. Lipping, surface porosity, osteophyte formation, eburnation, and percentage of joint surface affected were recorded on an ordinal scale and summed to create composite scores that were assigned a specific phase. Spearman's correlation indicated a positive relationship between each composite score and age (right shoulder = 0.752; left shoulder = 0.734). Transition analysis revealed a tendency toward earlier degeneration of the right shoulder. Bayesian statistics generated phase-related age estimates based on highest posterior density regions. Best age estimates were into the seventh decade at the 90th and 50th percentile. The proposed method supplements traditional techniques by providing age estimates beyond a homogenous 50+ age cohort.
PMID: 27930820 [PubMed - as supplied by publisher]
Estimating legal age based on fusion of The proximal humeral epiphysis.
Int J Legal Med. 2016 Dec 06;
Authors: Sánchez MB, Codinha S, García AM, Sánchez JA
The increasing demand which requires ascertaining the legal age of undocumented individuals who reach the various countries of the European Community means that new lines of research must be developed which help respond to questions posed by the Justice Administration. For this reason, this study has been designed on the basis of fusion times of the proximal humeral epiphysis. Moreover, the ultrasound scan has been used as the diagnostic method. It is a non-invasive technique, unlike the radiograph, which is used under current standards for the forensic diagnosis of age. Used as a study sample were the ultrasound images of the proximal humeral epiphysis among 221 individuals belonging to the Spanish population, of both genders, of ages ranging from 5 to 30 years. All of the images were classified into 6 stages of fusion based on the morphology of each. The results display differences among the six age groups proposed for each of the stages of fusion and are of great interest from the perspective of enforcing the Spanish Criminal Law Act on Minors, because Stage 4 would mean that the person being studied is under the age of 16 years in the case of males and 15 years in the case of females. These results, coupled with the use of ultrasound as a non-invasive diagnostic technique, make this study a very useful method when the use of radiographs is not possible.
PMID: 27924406 [PubMed - as supplied by publisher]
The effects of three different exercise modalities on markers of male reproduction in healthy subjects: a randomized controlled trial.
Reproduction. 2017 Feb;153(2):157-174
Authors: Hajizadeh Maleki B, Tartibian B, Chehrazi M
The aim of this study was to investigate the effects of moderate-intensity continuous training (MICT), high-intensity continuous training (HICT) and high-intensity interval training (HIIT) on markers of male reproduction including seminal markers of oxidative stress and inflammation as well as semen quality and sperm DNA integrity in healthy human subjects. A total of 397 healthy male volunteers were screened and 280 were randomly assigned to one of the MICT (n = 70), HICT (n = 70), HIIT (n = 70) and non-exercise (NON-EX, n = 70) groups. Subjects had inflammatory markers (IL-1β, IL-6, IL-8 and TNF-α), oxidants (ROS, MDA and 8-isoprostane), antioxidants (SOD, catalase and TAC), semen parameters and sperm DNA damage measured at baseline (T1), the end of week 12 (T2), the end of week 24 (T3), and 7 (T4) and 30 days (T5) after training. Chronic MICT, HICT and HIIT attenuated seminal markers of oxidative stress and inflammation with different kinetics for the three types of exercise (P < 0.05), and these changes were correlated with favorable improvements in semen quality parameters and sperm DNA integrity (P < 0.05). MICT was superior to HICT and HIIT in the improvements of markers of male reproductive function (P < 0.05). In conclusion, different exercise modalities favorably affect markers of male reproduction with different kinetics, suggesting intensity-, duration- and type-dependent adaptations to exercise training in healthy human subjects.
PMID: 27920258 [PubMed - in process]
Dental age estimation of children and adolescents: Validation of the Maltese Reference Data Set.
J Forensic Leg Med. 2016 Nov 28;45:29-31
Authors: Alsaffar H, Elshehawi W, Roberts G, Lucas V, McDonald F, Camilleri S
The purpose of this study was to validate a Reference Data Set, developed from the radiographic archives of the Dental Department, Mater Dei Hospital, Malta, for Dental Age Estimation of the Maltese population and to assess the accuracy of the method. Dental Panoramic Tomograms of 100 male and 100 females were selected. Tooth Development Stages were recorded for all 16 permanent teeth on the left and both third molars on the right. Summary and percentile data were calculated for each Tooth Development Stage and the Dental Age of each subject estimated using the unweighted average method. The distribution of the differences between the Dental and Chronological age was analysed. There was no statistically significant difference between the means of the Dental and Chronological ages and the mean differences were within acceptable limits. The conclusion was that the Dataset is valid for the Maltese population and the method used gives adequate accuracy.
PMID: 27918905 [PubMed - as supplied by publisher]
Sexual dimorphism of the tibia in contemporary Greek-Cypriots and Cretans: Forensic applications.
Forensic Sci Int. 2016 Nov 16;:
Authors: Kranioti EK, García-Donas JG, Almeida Prado PS, Kyriakou XP, Langstaff HC
Sex estimation is an essential step in the identification process of unknown heavily decomposed human remains as it eliminates all possible matches of the opposite sex from the missing person's database. Osteometric methods constitute a reliable approach for sex estimation and considering the variation of sexual dimorphism between and within populations; standards for specific populations are required to ensure accurate results. The current study aspires to contribute osteometric data on the tibia from contemporary Greek-Cypriots to assist the identification process. A secondary goal involves osteometric comparison with data from Crete, a Greek island with similar cultural and dietary customs and environmental conditions. Left tibiae from one hundred and thirty-two skeletons (70 males and 62 females) of Greek-Cypriots and one hundred and fifty-seven skeletons (85 males, 72 females) of Cretans were measured. Seven standard metric variables including Maximum length (ML), Upper epiphyseal breadth (UB), Nutrient foramen anteroposterior diameter (NFap), Nutrient Foramen transverse diameter (NFtrsv), Nutrient foramen circumference (NFCirc), Minimum circumference (MinCirc) and Lower epiphyseal breadth (LB) were compared between sexes and populations. Univariate and multivariate discriminant functions were developed and posterior probabilities were calculated for each sample. Results confirmed the existence of sexual dimorphism of the tibia in both samples as well as the pooled sample. Classification accuracy for univariate functions ranged from 78% to 85% for Greek-Cypriots and from 69% to 83% for Cretans. The best multivariate equations after cross-validation resulted in 87% for Greek-Cypriots and 90% accuracy for Cretans. When the samples were pooled accuracy reached 87% with over 95% confidence for about one third of the population. Estimates with over 95% of posterior probability can be considered reliable while any less than 80% should be treated with caution. This work constitutes the initial step towards the creation of an osteometric database for Greek-Cypriots and we hope it can contribute to the biological profiling and identification of the missing and to potential forensic cases of unknown skeletal remains both in Cyprus and Crete.
PMID: 27919515 [PubMed - as supplied by publisher]
Leo and Steve discuss Android meeting Gooligan, Windows Upgrades bypass Bitlocker, nearly one million UK routers taken down by a Mirai variant, the popular AirDroid app is "Doing it wrong", researchers invent a clever credit card disclosure hack, Cloudflare reports a new emerging botnet threat, deliberate backdoors discovered in 80 different models of Sony IP cameras, we get some closure on our SanFran MUNI hacker, a fun hack with Amazon's Echo and Google's Home, How to kill a USB port in seconds, a caution about keyless entry (and exit), too-easy-to-spoof fingerprint readers, an extremely troubling report from the UK, and finally some good news: the open-source covert USB hack defeating "BeamGun"!... plus a bunch of fun miscellany, some great Sci-Fi reader/listener book news, and... however many questions we're able to get to by the end of two hours!
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Hosts: Steve Gibson and Leo Laporte
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A test of the citrate method of PMI estimation from skeletal remains.
Forensic Sci Int. 2016 Nov 24;270:70-75
Authors: Wilson SJ, Christensen AM
Citrate content in bone has been shown to be associated with the postmortem interval (PMI), with citrate decreasing after death as a function of time. Here we test this method using porcine ribs for the period of 1-165days after death, and also assess citrate content and variation from samples placed into two different postmortem environments (terrestrial and aquatic). Higher citrate variation, lower citrate recovery, and a weaker association with time were found in this study as compared to others. Citrate content, however, was found to decrease with increasing PMI, and the method was found to be easy and inexpensive to apply. No significant differences were found in citrate loss between terrestrial and aquatic environments. Although more research is needed, citrate content appears to be a promising new approach in estimating PMI from skeletal remains.
PMID: 27915189 [PubMed - as supplied by publisher]
Euclidean Distances as measures of speaker similarity including identical twin pairs: A forensic investigation using source and filter voice characteristics.
Forensic Sci Int. 2016 Nov 17;270:25-38
Authors: San Segundo E, Tsanas A, Gómez-Vilda P
There is a growing consensus that hybrid approaches are necessary for successful speaker characterization in Forensic Speaker Comparison (FSC); hence this study explores the forensic potential of voice features combining source and filter characteristics. The former relate to the action of the vocal folds while the latter reflect the geometry of the speaker's vocal tract. This set of features have been extracted from pause fillers, which are long enough for robust feature estimation while spontaneous enough to be extracted from voice samples in real forensic casework. Speaker similarity was measured using standardized Euclidean Distances (ED) between pairs of speakers: 54 different-speaker (DS) comparisons, 54 same-speaker (SS) comparisons and 12 comparisons between monozygotic twins (MZ). Results revealed that the differences between DS and SS comparisons were significant in both high quality and telephone-filtered recordings, with no false rejections and limited false acceptances; this finding suggests that this set of voice features is highly speaker-dependent and therefore forensically useful. Mean ED for MZ pairs lies between the average ED for SS comparisons and DS comparisons, as expected according to the literature on twin voices. Specific cases of MZ speakers with very high ED (i.e. strong dissimilarity) are discussed in the context of sociophonetic and twin studies. A preliminary simplification of the Vocal Profile Analysis (VPA) Scheme is proposed, which enables the quantification of voice quality features in the perceptual assessment of speaker similarity, and allows for the calculation of perceptual-acoustic correlations. The adequacy of z-score normalization for this study is also discussed, as well as the relevance of heat maps for detecting the so-called phantoms in recent approaches to the biometric menagerie.
PMID: 27912151 [PubMed - as supplied by publisher]
In vitro effect of myo-inositol on sperm motility in normal and oligoasthenospermia patients undergoing in vitro fertilization.
Gynecol Endocrinol. 2016 Dec 2;:1-4
Authors: Artini PG, Casarosa E, Carletti E, Monteleone P, Di Noia A, Di Berardino OM
It is a known fact that abnormal seminal liquid specimens contain abnormal amounts of oxygen free radicals and reactive oxygen species (ROS), and that the use of antioxidant molecules both in vivo and in vitro leads to improvement of semen quality in terms of motility, reduction in DNA damage, with obvious consequences on the fertilization potential. Myo-inositol has been observed to have anti-oxidant properties and be present in much greater concentrations specifically in seminal liquid than in the blood. Moreover, there seems to be a direct relationship between myo-inositol and mitochondrial membrane potential (MMP) and sperm motility. Studies performed in vivo have demonstrated that a dietary supplementation with myo-inositol in men undergoing assisted reproduction techniques may improve sperm quality and motility in oligoasthenospermia (OAT) patients. In the following study we utilized myo-inositol in vitro to verify its effect on semen quality in both normal and OAT patients undergoing in vitro fertilization (IVF) with respect to standard sperm medium. In vitro incubation of seminal liquid carried out using myo-inositol (Andrositol-Lab, Lo.Li. Pharma-Roma, Italy) at a concentration of 15 μl/ml improved progressive motility in both normospermia and OAT subjects. In our opinion, myo-inositol may prove to be a useful strategy to improve sperm preparation for clinical use in IVF.
PMID: 27908215 [PubMed - as supplied by publisher]