In modern forensic practice, molecular genetic studies play an extremely important role, especially in wartime. DNA identification allows you to identify a person by traces of biological origin, which has become critically important for the identification of the dead and missing. The effectiveness of such studies largely depends on the choice of the optimal method of DNA extraction, which ensures the reliability and speed of results. In this regard, the choice of the most reliable, sensitive and resource-efficient method of DNA extraction becomes not only a technical but also a strategic task that directly affects the success of investigations and the reduction of the number of missing persons, which is extremely important in the humanitarian dimension of modern war. Aim. Comparative analysis of two commercial kits for manual DNA extraction — PrepFiler™ Forensic DNA Extraction Kit (Applied Biosystems) and NucleoSpin® DNA Forensic (Macherey-Nagel) — to determine the feasibility of their use for different types of biological samples and traces and categories of expert workload. Materials and methods. The study was conducted on the basis of the analysis of samples of trace biological material, in particular blood stains, saliva, tissue fragments and traces from the surfaces of objects. Two sets of reagents were used for DNA extraction: PrepFiler™ Forensic DNA Extraction Kit (Thermo Fisher Scientific) and NucleoSpin® DNA Forensic (Macherey-Nagel), 100 reactions each. DNA extraction was carried out according to the protocols provided by the manufacturers. The quality and quantity of the isolated DNA were determined by real-time polymerase chain reaction using the Applied Biosystems 7300 Real-Time PCR System. Further genotyping was performed by capillary electrophoresis on the Applied Biosystems 3500 Genetic Analyzer. Results. Comparative analysis showed that NucleoSpin® DNA Forensic is optimal for fast and convenient DNA isolation from simple reference samples such as saliva or blood, where no significant degradation or inhibitors are observed. It provides sufficient DNA concentration in a short time, however, in cases of working with complex or degraded samples (especially bone remains), partial STR profiles were more often observed. In contrast, PrepFiler™ and, in particular, PrepFiler™ BTA demonstrated better performance under such difficult conditions, providing complete profiles even in the presence of inhibitors or prolonged degradation. This makes it a reasonable choice for forensic cases with problematic samples, despite the higher cost. Conclusions. Molecular genetic research is a key tool in modern forensic practice, in particular for identifying a person from biological samples. The use of DNA identification methods based on polymerase chain reaction (PCR) ensures high accuracy and reliability of forensic examinations in criminal and civil cases, and also plays an important role in the identification of the deceased and missing, especially in wartime conditions. A comparative analysis of two commercial kits for manual DNA extraction — PrepFiler™ Forensic DNA Extraction Kit and NucleoSpin® DNA Forensic — showed that both kits have their advantages and can be effectively used depending on the type of biological material and the conditions of the examination. The choice of the optimal DNA extraction method significantly affects the quality and quantity of the obtained genetic material, which, in turn, determines the reliability of DNA profiling and the speed of the expert laboratory.

A Rare Case of Early Corneal Blood Staining After Post-operative Hyphema in a Child With Congenital Glaucoma and Haab’s Striae

Disseminated intravascular coagulation (DIC) is a potentially fatal condition commonly encountered in the context of obstetric complications. Various ocular manifestations have been reported as a consequence of DIC such as choroidal infarctions and vitreous and orbital hemorrhage. We report a case of a 24-year-old female, who developed DIC secondary to placental abruption at 33 weeks' gestation with a bilateral panophthalmitis-like picture, hyphema, and corneal blood staining.

BackgroundTo explore whether the application of mivacurium can facilitate laryngeal mask airway (LMA) insertion and benefit patients.MethodsA total of 167 patients undergoing hysteroscopy were randomly divided into mivacurium (group M) and control (group C) groups. The anesthesia induction scheme was mivacurium + sufentanil + propofol in group M, whereas mivacurium was replaced with saline in group C. The main outcome was the LMA insertion condition Secondary outcomes included attempts and elapsed time of LMA insertion, intraoperative anesthetic consumption, perioperative hemodynamics, postoperative sore throat (POST), nausea, vomiting, dizziness, and agitation.ResultsThere was no difference in the baseline data (p > 0.05). There was no significant difference in mouth opening; however, the incidence of swallowing, coughing, body movement, and pharyngeal spasm in group M was lower (p < 0.001), and the proportion of no resistance during LMA insertion was higher (80.5% vs. 21.2%, p < 0.001). The success rate of first-attempt LMA insertion in group M was higher (98.8% vs. 48.2%, p < 0.001), the elapsed time was shorter (16.9 (9.0) vs. 73.0 (91.5) s, p < 0.001), and fewer patients needed additional propofol (1.2% vs. 54.1%, p < 0.001). Blood staining on the LMA surface showed no significant difference, but the postoperative pharyngeal pain score in group M was lower (1.0 (1.0) vs. 2.0 (1.0), p < 0.001). Intraoperative propofol and remifentanil consumption, postoperative dizziness and nausea were lower in group M.ConclusionMivacurium facilitates LMA insertion and reduce intraoperative anesthetic consumption and adverse reactions, such as POST, nausea, and dizziness,so as to benefit the patient.Clinical Trial Registrationclinicaltrials.gov, identifier ChiCTR2500101122.

As a result of its viscous nature, blood stain patterns formed from blood splatter makes blood one of the most common pieces of evidence encountered. Bloodstain pattern analysis (BPA) is a branch of forensic science that examines the physical properties of blood and uses graphical pattern recognition to map out and evaluate bloodstains found at crime scenes and crime scene reconstruction. Crime scene reconstruction ensures a better understanding of events that unfolded at a scene. However, health and ethical issues in using real blood during reconstruction have led scientist to search for alternatives to mimic blood. An investigation using henna dye to mimic bloodstain by dropping from different heights (3 to 7 feet) was carried out in an outdoor crime scene simulation area. With the necessary steps involved in post scene processes diligently followed, it was observed that at 3 the formation of insignificant satellites with small spines around each drop, at 4 feet minimal satellite stains and longer spines appeared, at 5 feet very small spines and minimal satellites were observed with an elongation on parent droplet, at 6 feet shows more distant longer satellites and at 7 feet tiny elongated droplets observed with satellites and had smaller droplet attached. It was concluded that when the height of dropping fake blood is increased, the distance of satellite stains emerging from the fake bloodstain also increases. Therefore the bloodstain pattern formed is dependent on the height of the blood spatter on a horizontal surface. Therefore henna could be used to for investigations of crime scenes and educational studies on bloodstain pattern analysis (BPA).

The European Organization for External Quality Assurance Providers in Laboratory Medicine (EQALM) Haematology Working Group (WG) has previously provided two guidelines to support the standardization of haematology blood smear external quality assessment (EQA) (Vives CJL, Albarède S, Flandrin G, Heller S, Horvath K, Houwen B, etal. Guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear interpretation. Part I: control material. Clin Chem Lab Med 2004;42:922-6; Vives CJL, Van Blerk M, Albarède S, Gutierrez G, Heller S, Nordin G, etal. Guidelines for setting up an external quality assessment scheme for blood smear interpretation. Part II: survey preparation, statistical evaluation and reporting. Clin Chem Lab Med 2006;44:1039-43) but these recommendations did not include analytical performance specifications. In this paper the WG provides advice on the performance specifications for the evaluation of blood cell identification, comments on morphological characteristics and diagnostic hypothesis. To develop these specifications, the WG made a survey of the practices in use by EQALM members and provided a standard set of EQA data, provided by one of the members, for performance evaluation. The results of this exercise show a variation in the performance assessment outcomes from one EQA provider to another, suggesting that a degree of standardisation would be of benefit to participating laboratories. The WG has provided advice on common performance specifications, based on model 1 of the Milan Consensus (Sandberg S, Fraser CG, Horvath AR, Jansen R, Jones G, Oosterhuis W, etal. Defining analytical performance specifications: consensus statement from the 1st strategic conference of the European federation of clinical chemistry and laboratory medicine. Clin Chem Lab Med 2015;53:833-5).

Protocol for immunofluorescent staining, high-resolution imaging, and spatial projection of Schlemm's canal in mouse whole-mount corneal limbus tissue.

Synergistic protease-lipase treatment for enhanced blood stain removal from textiles: Process optimization and efficacy evaluation

The coupling of methylation-sensitive restriction enzyme with recombinant polymerase amplification and lateral flow dipstick for the detection of (menstrual) blood stains.

Human African trypanosomiasis is caused by protozoa of the genus Trypanosoma sp. and is primarily transmitted by the bite of the tsetse fly. This report presents a pediatric case diagnosed with human African trypanosomiasis in low-incidence areas of Central Africa, diagnosed by Cuban collaborators. Case presentation: A 12-year-old patient with a medical history had traveled to an area with active cases of trypanosomiasis within the last 2 years. The patient presented with recurrent fever, general weakness, and drowsiness. Rapid diagnostic tests were positive for salmonellosis and malaria. The patient was treated with antibiotics, with no clinical improvement. The diagnosis was positive for Trypanosoma sp. by thick blood smear and staining. Clinical improvement occurred after nifurtimox-eflornithine combination therapy according to the regional protocol for Trypanosoma brucei gambiense in the second phase of infection. Clinical improvement was seen at 48 hours. Conclusions: Human African trypanosomiasis is a common infection in regions where medical assistance is provided by Cuban medical personnel. The high endemism of other infections with similar symptoms and acquired immunity favors late identification without adequate knowledge and timely clinical-epidemiological analysis. A complementary definition and specific therapeutic measures ensure a good outcome and a better prognosis.

Forensics, along with the implementation of techniques from nanosciences, has brought about a change in the domain of forensic science and criminal investigation. It has brought about a positive change in the criminal investigation framework by making investigation more efficient and to-the-point. This paper reviews the nanotechnology-based techniques that are introduced in forensic science to unravel the mysteries behind crimes as it highlights various kinds of nano-sized particles, nanodevices along with their applications that the forensic experts use to analyze the evidences collected from the crime scenes. Google scholar, PubMed, and Scopus search engines are used to select the related articles to write this review paper. DNA analysis, fingerprint detection, drug detection, explosive analysis, blood stain analysis, time since death estimation, and analysis of counterfeit documents are some of the sectors in which nanotechnology has made notable contributions. As the paper unfolds, it makes sure that the readers get the taste of both the worlds, and that it helps them grasp the concept and idea behind the techniques used to make a change across the globe.

A robust dataset for surgical instrument detection to aid autonomous robotic surgery.

The use of epigenetic clocks for measuring age acceleration in the field of cancer research has been a common practice for many years. In forensic genetics, DNA methylation can be used to estimate the age of a stain donor. As lifestyle and disease can alter a person’s methylation profile, the accuracy of forensic age estimation tools might decrease compared to the chronological age when estimating a person affected by cancer. In our study, we applied the VISAGE enhanced age estimation tool on blood samples from cancer patients suffering from a variety of cancer entities, including solid and hematologic tumours. A comparison of the age estimation errors between the cancer patients (n = 100) and a healthy control cohort (n = 102) revealed small statistically significant differences and a tendency towards age acceleration in the blood of these patients. Although this study showed that in patients with aggressive cancers (like CLL or AML) estimation accuracy is clearly decreased, for most entities the observed differences were subtle and an analysis of individual CpG sites did not reveal strikingly different methylation patterns. Conclusively, age estimation on blood stains from cancer patients might not result in significantly higher estimation errors, except for very aggressive forms of cancer.

Investigating handheld near-infrared spectroscopy for forensic body fluid analysis.

Laceyella sacchari strain BK-TM, isolated from the Salt Lake of Chott Ech Chergui (El-Bayadh, Algeria), was found to produce an extracellular thermostable serine protease (SPLS). The highest level of protease activity detected after 6days of incubation at 50°C was 44,600U/mL. SPLS was purified after heat treatment for 5min at 80°C, subjected to ammonium sulfate fractionation (80%), and Sephacryl S-200 High Resolution (HR) column purification. The purified enzyme consists of a single protein with an approximate molecular weight of 29kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The sequence of the 24 N-terminal residues of SPLS was highly similar to those of Thermoactinomycetaceae proteases. Furthermore, the complete inhibition by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DIFP) indicates that SPLS is a member of the serine protease family. The enzyme exhibits optimal activity at pH 9 and 70°C. The thermostability of SPLS increased when 1mM of Ca2+ was added, with a half-life of 8h at 80°C and 3h at 90°C. Its catalytic activity was superior to that of SPSM from Streptomyces mutabilis strain TN-X30, PREFERENZ P300, and PROTEASE Type XIV. Interestingly, SPLS demonstrated high compatibility with ISIS and Maison Det, serving as solid and liquid laundry detergents, respectively. Furthermore, performance evaluations revealed that SPLS effectively removes blood stains. Overall, SPLS exhibited interesting biochemical features, suggesting its potential use as a cleaning bio-additive in detergent formulations.

ObjectivesThis study evaluated the efficacy of three distinct bleaching agents over time on blood-stained, devitalized teeth. Furthermore, the recoloring subsequent to bleaching will be monitored.MethodsThe study was conducted on 60 caries-free, unfilled, upper human incisors. The Freccia and Peters blood staining technique was employed, and four groups (n = 15) were identified: control, 35% hydrogen peroxide-treated, 37% carbamide peroxide-treated, and sodium perborate-treated groups. Color differences were measured using ΔE00, ΔWID, L*, a*, and b* values. To investigate tooth discoloration after bleaching, 10 unbleached teeth with three groups of 10 bleached teeth were compared by vine staining. The group of bleached teeth was restored immediately, another group waited one week, and the third group had sodium ascorbate applied and analyzed using one-way analysis of variance tests (p < 0.05).ResultsAmong the groups, carbamide peroxide exhibited the most significant whitening during the 6-day bleaching process, followed by hydrogen peroxide and sodium perborate. Subsequent examination of the wine recoloring of post-bleaching samples demonstrated that bleached teeth exhibited a heightened propensity for recoloration in contrast to unbleached teeth. Notably, sodium ascorbate treatments for hydrogen peroxide neutralization and the wait-and-restore approach were not statistically significant in terms of preventing recoloration.ConclusionsSodium perborate is less effective and more time-consuming than hydrogen peroxide or carbamide peroxide for bleaching purposes. Carbamide peroxide is the most effective bleaching agent. The sodium ascorbate treatment and the wait-and-restore approach are ineffective in preventing recoloring. Bleached teeth have more discoloration than unbleached teeth.

Japanese traditional seasoning "miso" accelerates alteration of bloodstain color: Proof of innocent for a murder committed in 1966.

Blood stains can be used to understand the dynamics of a crime, to verify the identity of a suspect, to perform pattern analysis in reconstructing the incident, or to determine the time when a crime was committed. Changes in the cell and cell membrane surfaces of erythrocytes, which are blood cells, can be indicators for quantitative assessment of the time of the crime. Atomic force microscopy (AFM) is a rapidly developing tool that has recently been used to assess the age of blood stains and has the potential to provide useful information for forensic investigation. In this study, erythrocytes were observed to exhibit morphological changes in the cell surface over time using AFM and cellular viscoelasticity through force-distance curve measurements. Approximately, 80 natural erythrocytes were measured in the experiments. Topographic measurements were made for each erythrocyte. It was observed that the biconcave shapes of erythrocytes deteriorated over time and that there were differences in the cross-sectional planes of the cell membrane. As a result, it was observed that the shapes of erythrocytes deformed over time. In our study, images that can determine the changes in the morphological properties of erythrocytes were successfully obtained, and cell morphological properties and cell viscoelasticity could be observed with AFM. On the 1st day, the deepest part of the erythrocytes was 354.50 nm on average, 176.01 nm on the 7th day, 116.31 nm on the 14th day, and 56.99 nm on the 30th day. Accordingly, Young's modulus (Force curve analysis) was measured to check the stresses occurring on the membrane surfaces. On the 1st day, the young's modulus of the erythrocyte membrane was 184.20 MPa on average, 345.57 MPa on the 7th day, 763.48 MPa on the 14th day, and 1631.82 MPa on the 30th day. The results of this study showed the effectiveness of AFM in detecting structural and morphological changes that occur during the aging process of erythrocytes.

A study was conducted to analyse the histological structure of lymphocytes and monocytes in dogs with dermatitis before and after therapy. Dermatitis is skin inflammation that can be caused by bacterial infections, allergies, or environmental factors. This study used 20 dogs with dermatitis that were given a combination of antiparasitic, antibiotic, anti-inflammatory, and antihistamine therapy. Blood samples were taken before and after therapy to be analysed using Morphology of Edge Blood (MDT) staining to observe changes in the histological structure of lymphocyte and monocyte cells. The results showed: before therapy, lymphocytes and monocytes experienced more necrosis (P&lt;0.05) histological structure changes such as pycnosis, karyorexis, and karyolysis. After therapy, there was a decrease in the number of cells that experienced necrosis in lymphocytes, from 8.45 ± 2.08% to 6.85 ± 1.75%. Whereas in monocytes, there was a decrease in the number (P&lt;0.05) of necrotised cells at post- therapy, from 27.5±19.4% to 13.5±10.3%. The conclusion of this study is that the histological structure of lymphocytes and monocytes that experience necrosis is less than cells that do not necrose. The histological structure of the cells showed a lot more normal. The combination therapy used proved effective in improving the structure of lymphocytes and monocytes.

Purification, characterization, structural elucidation, and industrial applications of thermostable alkaline protease produced by seaweed-associated Nocardiopsis dassonvillei strain VCs-4.