Persistence Of DNA Research Articles

A synthetic fingerprint solution and its importance in DNA transfer, persistence and recovery studies

A review of the literature on DNA transfer and persistence highlights many difficulties that are encountered when conducting research of this nature. One of the main problems highlighted repeatedly in the literature is the prevalence of inherent uncontrolled variation that accompany these studies, and in turn, the results obtained. This work aims to decrease the amount of intrinsic variability associated with DNA transfer and persistence experiments using a realistic proxy solution which is adaptable, of known composition, reproducible, and capable of being standardised. This proxy is composed of three parts: a synthetic fingerprint solution, cellular DNA, and cell free DNA. In this proof-of-concept study the proxy was tested with a small-scale DNA transfer and recovery experiment and the data obtained suggests that the use of a solution that mimics real fingerprint secretions, over an alternative (such as buffer or a body fluid), is important when working with non-donor provided trace DNA samples. This is because the DNA deposit solution likely impacts the transfer of DNA from fingers/hands to a surface as well as the ability to recover the biological material once deposited.

An optically fabricated gradient nanochannel array to access the translocation dynamics of T4-phage DNA through nanoconfinement.

It has been widely recognized that nanostructures in natural biological materials play important roles in regulating life machinery. Even though nanofabrication techniques such as two-photon polymerization (TPP) provide sub-100 nm fabrication resolution, it remains technologically challenging to produce 3D nanoscale features modeling the complexity in vivo. We herein demonstrate that a nanochannel array carrying different sizes and nanostructures with gradually transitioning dimensions can be easily produced on a slightly tilted nano-stage. Using the gradient nanochannel array, we systematically investigate the factors affecting the dynamics of DNA translocation through nanoconfinement, including the size of biomolecules and geometrical features of the physical environment, which resembles the selectivity of nanopores in the cell membrane. It is observed that T4-phage DNA shows distinctive conformational transition dynamics during translocation through nanochannels driven by electric field or flow, and the deformation energy required for DNA to enter the nanochannels depends on both chemical environmental conditions, i.e., the ionic strength regulating DNA persistence length and nanochannel dimension. In the electric field, DNA repeatedly gets stretched and compressed during its migration through the nanochannel, reflected by elevated fluctuation in extension, which is substantially greater than the thermal fluctuation. However, driven by flow, DNA remains stretched during translocation through nanochannels, and shows variances in extension of merely a few hundred nanometers. These results indicate that the optically fabricated gradient nanochannel array is a suitable platform for optimizing the experimental conditions for biomedical applications such as gene mapping, and verify that production of complex three dimensional (3D) nanostructures can be greatly simplified by including slight inclination during TPP fabrication.

Discovery of a first-in-class orally available HBV cccDNA inhibitor

The persistence of covalently closed circular DNA (cccDNA) in infected hepatocytes is the major barrier preventing viral eradication with existing therapies in patients with chronic hepatitis B. Therapeutic agents that can eliminate cccDNA are urgently needed to achieve viral eradication and thus HBV cure. A phenotypic assay with HBV-infected primary human hepatocytes (PHHs) was employed to screen for novel cccDNA inhibitors. A HBVcircle mouse model and a uPA-SCID (urokinase-type plasminogen activator-severe combined immunodeficiency) humanized liver mouse model were used to evaluate the anti-HBV efficacy of the discovered cccDNA inhibitors. Potent and dose-dependent reductions in extracellular HBV DNA, HBsAg, and HBeAg levels were achieved upon the initiation of ccc_R08 treatment two days after the HBV infection of PHHs. More importantly, the level of cccDNA was specifically reduced by ccc_R08, while it did not obviously affect mitochondrial DNA. Additionally, ccc_R08 showed no significant cytotoxicity in PHHs or in multiple proliferating cell lines. The twice daily oral administration of ccc_R08 to HBVcircle model mice, which contained surrogate cccDNA molecules, significantly decreased the serum levels of HBV DNA and antigens, and these effects were sustained during the off-treatment follow-up period. Moreover, at the end of follow-up, the levels of surrogate cccDNA molecules in the livers of ccc_R08-treated HBVcircle mice were reduced to below the lower limit of quantification. We have discovered a small-molecule cccDNA inhibitor that reduces HBV cccDNA levels. cccDNA inhibitors potentially represent a new approach to completely cure patients chronically infected with HBV. Covalently closed circular DNA (cccDNA) persistence in HBV-infected hepatocytes is the root cause of chronic hepatitis B. We discovered a novel small-molecule cccDNA inhibitor that can specifically reduce cccDNA levels in HBV-infected hepatocytes. This type of molecule could offer a new approach to completely cure patients chronically infected with HBV.

Reporting on forensic biology findings given activity level issues in the Netherlands

There appears to be some hesitation within the forensic biology community to formally evaluate and report on findings given activity level propositions. This hesitance in part stems from concerns about the lack of relevant data on the dynamics of biological traces and doubt about the relevance of such expert opinions to the trier of fact. At the Netherlands Forensic Institute formal evaluative opinions on the probability of case findings given propositions at the activity level are provided since 2013, if requested by a mandating authority. In this study we share the results from a retrospective analysis of 74 of such requests. We explore which party initiates requests, the types of cases that are submitted, the sources of data being used to assign probabilities to DNA transfer, persistence, prevalence and recovery (TPPR) events, the conclusions that were drawn by the scientists, and how the conclusions were used by the courts. This retrospective analysis of cases demonstrates that published sources of data are generally available and can be used to address DNA TPPR events in most cases, although significant gaps still remain. The study furthermore shows that reporting on forensic biology findings given activity level propositions has been generally accepted by the district and appeal courts, as well as the other parties in the criminal justice system in the Netherlands.

How changes to the substrate’s physical characteristics can influence the deposition of touch and salivary deposits

An in-depth study into the physical substrate characteristics such as substrate surface roughness, topography, and physicochemical characteristics like wettability and surface free energy (SFE) was conducted to investigate the impact on the deposition and adherence of touch and salivary deposits on aluminium and polypropylene. A robust protocol was established to generate a set of substrates with a controlled linear surface roughness range (0.5–3.5 µm) in order to identify the impact of surface roughness on DNA transfer, persistence, prevalence, and recovery (DNA-TPPR). The polypropylene substrate was shown to produce fibres when artificially roughened, becoming more prominent at a higher surface roughness range, and has shown to have a direct impact on the distribution of salivary and touch deposits. At the low to moderate surface roughness range 0.5–2.0 µm, salivary and touch deposits have generally shown to follow the topographical features of the substrate they were deposited on, before a plateau of the surface roughness measure on the deposit was observed, indicating that a saturation point was reached and the grooves in the substrate were beginning to fill. Touch deposits have shown to maintain a consistent deposition height pre-surface roughness threshold, irrespective of substrate surface roughness while the deposition height of salivary deposits was heavily influenced by substrate surface roughness and topography. The substrate SFE, wettability, hydrophobicity, and the surface tension of the deposit was shown to drive the adhesion properties of the saliva and touch deposits on the respective substrates, and it was observed that this may be of importance for the improvement of the current DNA-TPPR understanding, DNA sampling protocols, and DNA transfer considerations within casework.

Expression of Concern: DNA Persistence and Relapses Questions on the Treatment Strategies of Enterococcus Infections of Prosthetic Valves.

Expression of Concern: DNA Persistence and Relapses Questions on the Treatment Strategies of Enterococcus Infections of Prosthetic Valves.

An investigation of DNA damage and DNA repair in chemical carcinogenesis triggered by small-molecule xenobiotics and in cancer: Thirty years with the comet assay

In the present review we addressed the determination of DNA damage induced by small-molecule carcinogens, considered their persistence in DNA and mutagenicity in in vitro and in vivo systems over a period of 30 years. The review spans from the investigation of the role of DNA damage in the cascade of chemical carcinogenesis. In the nineties, this concept evolved into the biomonitoring studies comprising multiple biomarkers that not only reflected DNA/chromosomal damage, but also the potential of the organism for biotransformation/elimination of various xenobiotics. Since first years of the new millennium, dynamic system of DNA repair and host susceptibility factors started to appear in studies and a considerable knowledge has been accumulated on carcinogens and their role in carcinogenesis. It was understood that the final biological links bridging the arising DNA damage and cancer onset remain to be elucidated. In further years the community of scientists learnt that cancer is a multifactorial disease evolving over several decades of individual´s life. Moreover, DNA damage and DNA repair are inseparable players also in treatment of malignant diseases, but affect substantially other processes, such as degeneration. Functional monitoring of DNA repair pathways and DNA damage response may cast some light on above aspects. Very little is currently known about the relationship between telomere homeostasis and DNA damage formation and repair. DNA damage/repair in genomic and mitochondrial DNA and crosstalk between these two entities emerge as a new interesting topic.

Mechanical Constraint Effect on DNA Persistence Length.

Persistence length is a significant criterion to characterize the semi-flexibility of DNA molecules. The mechanical constraints applied on DNA chains in new single-molecule experiments play a complex role in measuring DNA persistence length; however, there is a difficulty in quantitatively characterizing the mechanical constraint effects due to their complex interactions with electrostatic repulsions and thermal fluctuations. In this work, the classical buckling theory of Euler beam and Manning's statistical theories of electrostatic force and thermal fluctuation force are combined for an isolated DNA fragment to formulate a quantitative model, which interprets the relationship between DNA persistence length and critical buckling length. Moreover, this relationship is further applied to identify the mechanical constraints in different DNA experiments by fitting the effective length factors of buckled fragments. Then, the mechanical constraint effects on DNA persistence lengths are explored. A good agreement among the results by theoretical models, previous experiments, and present molecular dynamics simulations demonstrates that the new superposition relationship including three constraint-dependent terms can effectively characterize changes in DNA persistence lengths with environmental conditions, and the strong constraint-environment coupling term dominates the significant changes of persistence lengths; via fitting effective length factors, the weakest mechanical constraints on DNAs in bulk experiments and stronger constraints on DNAs in single-molecule experiments are identified, respectively. Moreover, the consideration of DNA buckling provides a new perspective to examine the bendability of short-length DNA.

Persistence of DNA from canine parvovirus modified-live virus in canine tissues.

Canine parvovirus (CPV-2) modified-live virus vaccine strain can replicate in lymphoid tissues and intestinal mucosa after administration, being shed through canine faeces. Detection of vaccine strains has been reported in the bloodstream and faeces, potentially interfering with molecular diagnostic tests. The persistence of these strains in canine tissues has not yet been described. With this aim, canine tissues were tested during a molecular survey to screen for the presence of canine enteric viruses. Tissue samples from 165 dead dogs were tested by a conventional PCR assay. Positive samples and five commercial vaccines were subjected to sequence analysis. Vaccinal strains were detected and virus load was measured by using a set of real-time PCR assays using minor-groove binder (MGB) probes. Seventy-five dogs (45.4%) tested positive for CPV-2. Strains from 70 dogs were characterised as field variants. The presence of CPV sequences of vaccine origin was observed in the spleen, intestine, and mesenteric lymph nodes of five young dogs. Vaccinal strains were detected from 12 to 24 days after the last vaccine administration. Viral loads comprised between 6.3 × 102 and 9.95 × 104 DNA copies/10 µl of template. This study confirms that CPV vaccinal strains can be detected in canine tissues after vaccination, so post-mortem diagnosis of CPV infection needs further molecular analyses to assess the viral type (vaccine or field strains). The present study updates the current information on the persistence of CPV vaccine strains in canine tissues and their possible interference with molecular assays.

Safety and Outcomes of a Plasmid DNA Vaccine Encoding the ERBB2 Intracellular Domain in Patients With Advanced-Stage ERBB2-Positive Breast Cancer

High levels of ERBB2 (formerly HER2)-specific type 1 T cells in the peripheral blood are associated with favorable clinical outcomes after trastuzumab therapy; however, only a minority of patients develop measurable ERBB2 immunity after treatment. Vaccines designed to increase ERBB2-specific T-helper cells could induce ERBB2 immunity in a majority of patients. To determine the safety and immunogenicity of 3 doses (10, 100, and 500 μg) of a plasmid-based vaccine encoding the ERBB2 intracellular domain (ICD). Single-arm phase 1 trial including 66 patients with advanced-stage ERBB2-positive breast cancer treated in an academic medical center between 2001 and 2010 with 10-year postvaccine toxicity assessments. Data analysis was performed over 2 periods: January 2012 to March 2013 and July 2021 to August 2022. Patients were sequentially enrolled to the 3 dose arms. The vaccine was administered intradermally once a month with soluble granulocyte-macrophage colony-stimulating factor as an adjuvant for 3 immunizations. Toxicity evaluations occurred at set intervals and yearly. Peripheral blood mononuclear cells were collected for evaluation of immunity. Biopsy of vaccine sites at weeks 16 and 36 measured DNA persistence. Safety was graded by Common Terminology Criteria for Adverse Events, version 3.0, and ERBB2 ICD immune responses were measured by interferon-γ enzyme-linked immunosorbent spot. Secondary objectives determined if vaccine dose was associated with immunity and evaluated persistence of plasmid DNA at the vaccine site. A total of 66 patients (median [range] age, 51 [34-77] years) were enrolled. The majority of vaccine-related toxic effects were grade 1 and 2 and not significantly different between dose arms. Patients in arm 2 (100 μg) and arm 3 (500 μg) had higher magnitude ERBB2 ICD type 1 immune responses at most time points than arm 1 (10 μg) (arm 2 compared with arm 1, coefficient, 181 [95% CI, 60-303]; P = .003; arm 3 compared with arm 1, coefficient, 233 [95% CI, 102-363]; P < .001) after adjusting for baseline factors. ERBB2 ICD immunity at time points after the end of immunizations was significantly lower on average in patients with DNA persistence at week 16 compared with those without persistence. The highest vaccine dose was associated with the greatest incidence of persistent DNA at the injection site. In this phase 1 nonrandomized clinical trial, immunization with the 100-μg dose of the ERBB2 ICD plasmid-based vaccine was associated with generation of ERBB2-specific type 1 T cells in most patients with ERBB2-expressing breast cancer, and it is currently being evaluated in randomized phase 2 trials. ClinicalTrials.gov Identifier: NCT00436254.

African trypanosome infection patterns in cattle in a farm setting in Southern Ghana

African Trypanosomiasis is a debilitating disease in both humans and animals that occurs in sub-Saharan Africa and has a severe negative impact on the livelihood of people in the affected areas. The disease is caused by protozoan parasites of the genus Trypanosoma, which is often described simply as blood-borne; however, a number of studies have shown the parasite inhabits many different environments within the host. Control of the disease involves measures that include the use of trypanocidal drugs to which there are growing number of reported cases of resistance. Here, the patterns of trypanosome DNA presence during a diminazene aceturate treatment round on a cohort of cattle in Adidome, Ghana were assessed. A group of 24 cows were selected irrespective of age and sex and the infecting trypanosome species followed for 18 days before and after treatment with diminazene aceturate in the blood and skin of the animals using a diagnostic nested PCR that targeted the alpha-beta tubulin gene array. Persistence of trypanosome DNA was observed over the period and parasite DNA was readily detected in both the skin and blood, with parasite DNA disappearing and reappearing in both across the study. Moreover, there was limited correlation between the parasite DNA detected in the skin and blood. Overall, the data show the patterns of a natural trypanosome infection during drug treatment. In addition, the diagnostic potential of sampling the skin for African trypanosomiasis is highlighted.

DNA on the inside door handles of entrance doors

DNA from door handles on entry doors could provide a clue as to who last left the scene. However, after years of extensive research on DNA transfer and persistence it can be considered common knowledge that general claims like "the last who touched leaves the most DNA" do not hold true. But who's DNA do we find on door handles that are usually used several times per day by the inhabitants? To assess this question, we sampled inside door handles from real-life burglaries and at the same time collected reference samples from all the inhabitants, to determine if we can detect any (major) profiles from non-inhabitants. We also searched to evaluate how often we detect DNA from the person who last touched the door handle as a (major) contributor. Only small amounts of DNA were recovered from the handles, originating most often, but not always, from inhabitants or even the last inhabitant touching the handle.

DNA on drugs

The use of illicit drugs is a continuing blight on society. Detecting DNA from individuals involved in the manufacturing and distribution of drugs can provide valuable investigative information or strategic intelligence which, in turn, can be used to disrupt the supply and distribution of illicit drugs. Our study details the transfer, persistence, prevalence, and recovery of human DNA on the exterior of tablets and capsules, as well as within drug powders. Various experiments were conducted to mimic stages in the creation and packaging of tablets and capsules. We showed that the act of brief contact (1–3 s) is sufficient to generate informative DNA profiles that can be uploaded and compared to databases internationally. This work complements chemical drug profiling data by linking seizures to each other and individuals via DNA profiles, providing information to prosecution or intelligence agencies. The generation of DNA information from illicit drug preparations is another tool that can be used in the fight against illicit drug manufacture and distribution.

Influence of temperature and water quality on the persistence of human mitochondrial DNA, human Hf183 Bacteroidales, fecal coliforms and enterococci in surface water in human fecal source tracking context.

Mitochondrial DNA (mtDNA) is used as a genetic marker to track fecal contamination in surface water. Its potential to effectively discriminate between the nonpoint sources of fecal pollution (e.g. human, livestock) in water environments is relevant for water quality management. However, there is a lack of knowledge about the environmental persistence of mtDNA in relation to those of other microbial parameters, such as fecal indicator bacteria (FIB). In this study, mesocosms composed of water collected from four rivers and tap water were spiked with raw wastewater to mimic human fecal contamination. Mesocosms composed of raw wastewater were also studied. The mesocosms were incubated at 4 °C or at 22 °C for 189 days, from which the levels of human mtDNA (HumtDNA) and human Bacteroidales (Hf183) were measured by qPCR. The levels of FIB (fecal coliforms and enterococci) and heterotrophs were determined by culture methods along with the determination of physicochemical attributes. The decay rates of the genetic markers and FIB were determined with first-order decay rate models. The decay rates of HumtDNA (0.004-0.059 d-1), Hf183 (0.007-0.082 d-1), and the two FIBs (0.005-0.066 d-1) were similar at 4 °C, while the genetic markers both had higher decay rates (0.013-0.919 d-1) at 22 °C. Different HumtDNA decay rates were observed between the river mesocosms (0.043-0.919 d-1) and the wastewater and tap water mesocosms (0.004-0.095 d-1). Covariations of pH and conductivity among the HumtDNA, Hf183 and FIB decay rates were observed. HumtDNA and Hf183 had similar environmental persistence, whereas fecal coliforms and enterococci persisted longer at 22 °C. Finally, HumtDNA had the same trends of persistence in the four river mesocosms, suggesting a relative stability of this marker in different rivers. Our results suggest that HumtDNA could be more suitable for tracking the source of a recent fecal contamination in complement to FIB.

Environmental DNA Metabarcoding: A Novel Contrivance for Documenting Terrestrial Biodiversity.

Simple SummaryThe innovative concept of environmental DNA has found its foot in aquatic ecosystems but remains an unexplored area of research concerning terrestrial ecosystems. When making management choices, it is important to understand the rate of eDNA degradation, the persistence of DNA in terrestrial habitats, and the variables affecting eDNA detectability for a target species. Therefore an attempt has been made to provide comprehensive information regarding the exertion of eDNA in terrestrial ecosystems from 2012 to 2022. The information provided will assist ecologists, researchers and decision-makers in developing a holistic understanding of environmental DNA and its applicability as a biodiversity monitoring contrivance.The dearth of cardinal data on species presence, dispersion, abundance, and habitat prerequisites, besides the threats impeded by escalating human pressure has enormously affected biodiversity conservation. The innovative concept of eDNA, has been introduced as a way of overcoming many of the difficulties of rigorous conventional investigations, and is hence becoming a prominent and novel method for assessing biodiversity. Recently the demand for eDNA in ecology and conservation has expanded exceedingly, despite the lack of coordinated development in appreciation of its strengths and limitations. Therefore it is pertinent and indispensable to evaluate the extent and significance of eDNA-based investigations in terrestrial habitats and to classify and recognize the critical considerations that need to be accounted before using such an approach. Presented here is a brief review to summarize the prospects and constraints of utilizing eDNA in terrestrial ecosystems, which has not been explored and exploited in greater depth and detail in such ecosystems. Given these obstacles, we focused primarily on compiling the most current research findings from journals accessible in eDNA analysis that discuss terrestrial ecosystems (2012–2022). In the current evaluation, we also review advancements and limitations related to the eDNA technique.

The Use of Ellagic Acid and Annona Muricata Improves Semen Quality in Men with High-Risk Papillomavirus Infection.

Background: Few data are currently available on the treatment of patients with HPV infection. In particular, there is no agreement on the use of antioxidants in these patients. Ellagic acid and annona muricata appear to improve HPV clearance in infected women. However, it is presently unknown whether they could enhance the clearance of HPV infection in infertile male patients. Aim: To evaluate the effects of a commercially available combined compound containing ellagic acid and annona muricata on semen quality in patients with documented papillomavirus (HPV) infection, and on the frequency of HPV DNA detection in seminal fluid after treatment. In addition, anti-sperm antibodies and the percentage of spermatozoa with fragmented DNA were evaluated. Materials and methods: This was a retrospective case-control study including patients attending our center for infertility. Fifty selected patients who were positive for high risk (HR)-HPV with available semen analysis results were consecutively enrolled. Patients were classified into two groups, according to the clinician’s decision to either administer ellagic acid 100 mg and annona muricata 100 mg (combined tablet formulation) for a period of three months (Group A; 25 patients), or to re-evaluate HPV DNA after a period of active surveillance only (protected sexual intercourse) (Group B; 25 patients). Results: Group A patients had a mean age of 31.0 ± 11.0 years, while Group B was 33.0 ± 8.0 years old (p > 0.05). After three months of treatment with ellagic acid and annona muricata, all conventional seminal parameters improved more significantly in Group A than in Group B patients: sperm concentration = 45 mil/mL vs. 20 mil/mL (p < 0.05); sperm progressive motility = 45% vs. 18% (p < 0.05); and normal sperm morphology = 18% vs. 6% (p < 0.05). After the treatment, the frequency of persistence of HPV DNA in the seminal fluid was significantly lower in Group A patients compared to those in Group B (12/25 = 48% vs. 22/25 = 88%; p < 0.05). Finally, after 3 months, Group A showed a significant reduction in anti-sperm antibodies and in the percentage of spermatozoa with fragmented DNA. Conclusion: The results of this study demonstrate, for the first time, the effects of a commercially available combined compound containing ellagic acid and annona muricata on semen quality in patients with HR-HPV infection, and that this therapy is also associated with a significant reduction in the persistence of HPV DNA in the seminal fluid.

Global burden of occult hepatitis B virus infection: where we are and what to do

Occult hepatitis B virus (HBV) infection is characterised by the persistence of replication-competent HBV DNA in the liver (with or without HBV DNA in the blood) of individuals testing negative for HBsAg, using currently available assays.1 Occult HBV infection represents a phase of the natural history of HBV infection,2 in which HBV DNA in the form of a covalently closed circular DNA minichromosome has suppressed replicative activity and gene expression with extremely low rates of virion production.

Detection and persistence of environmental DNA (eDNA) of the different developmental stages of a vector mosquito, Culex pipiens pallens.

Preventing mosquito-borne infectious diseases requires that vector mosquitoes are monitored and controlled. Targeting immature mosquitoes (eggs, larvae, and pupae), which have less mobility than adults, is an effective management approach. However, conducting these surveys is often difficult due to the limitations of morphological classification and survey costs. The application of environmental DNA (eDNA) analysis can solve these issues because it allows easy estimation of species distribution and morphology-independent species identification. Although a few previous studies have reported mosquito eDNA detection, there is a gap in knowledge regarding the dynamics related to the persistence of immature mosquito eDNA. We used Culex pipiens pallens, a vector of West Nile fever, as a model species. First, we developed a species-specific detection assay and confirmed its specificity using in silico and in vitro tests. Next, we conducted laboratory experiments using breeding tanks. Water samples were collected at each developmental stage. In addition, water samples were collected daily until the seventh day after emergence from the pupae. We quantified eDNA using real-time PCR with the developed assay to investigate the dynamics of mosquito eDNA. The specificity of the developed assay was confirmed by in silico and in vitro tests. Mosquito eDNA was detected at all developmental stages and detected up to seven days after emergence of pupae. In particular, high concentrations of eDNA were detected immediately after hatching from eggs and after emergence from pupae. Highly frequent positive eDNA signals were continuously detected between egg hatching and pupa hatching. Mosquito eDNA was detected immediately after the eggs were introduced, and eDNA-positive detections continued until pupae emergence, suggesting that eDNA analysis is useful for monitoring mosquito larvae. In the future, monitoring immature mosquitoes using eDNA analysis will contribute to prevent mosquito-borne infectious diseases.

Effects of total dissolved gas supersaturation and sediment on environmental DNA persistence of grass carp (Ctenopharyngodon idella) in water

Environmental DNA (eDNA) technology has become an alternative tool for monitoring aquatic communities due to its sensitive, economical, and non-invasive properties. However, the application of this technique is often limited by the complexity of environmental conditions, which often poses a barrier to the transmission of biological information. Here, we conducted a series of experiments with grass carp as the target species to evaluate the effects of total dissolved gas (TDG) supersaturation and sediment on the persistence of eDNA under different flow conditions. The results showed TDG supersaturation promoted eDNA decay in still water but with no significant effect in flowing water for rapid dissipation of TDG. For sediment, its presence accelerated the decay of eDNA no matter the flow conditions. The grass carp eDNA showed an exponential decay pattern in water and the decay rate constant decreased gradually with time. Our study highlights the importance of integrating experimental results with the natural environment and provides an important reference for species monitoring using eDNA technology in aquatic ecosystems with high dams built.

Pooled safety analysis of STRATA2011 and STRATA2016 clinical trials evaluating the use of StrataGraft® in patients with deep partial-thickness thermal burns

ObjectiveThis analysis includes pooled safety data from 2 clinical trials (NCT01437852; NCT03005106) that evaluated the safety and efficacy of StrataGraft in patients with deep partial-thickness (DPT) burns. MethodsThe study enrolled 101 adult patients with thermal burns covering 3–49% of total body surface area. Patients were followed for up to 1 year. The pooled safety events included: adverse events (AEs), adverse reactions (ARs), serious AEs (SAEs), discontinuation, and deaths; immunological responses (reactivity to panel reactive antibodies [PRA] and human leukocyte antigen [HLA] class 1 alleles); and persistence of allogeneic DNA from StrataGraft. ResultsEighty-seven (86.1%) patients experienced 397 AEs. Thirty patients (29.7%) experienced ARs; 16 patients (15.8%) experienced SAEs. The most frequent AEs were pruritus (n = 31; 30.7%), and blister, hypertension, and hypertrophic scar (n = 11 each; 10.9%); the most common AR was pruritus (n = 13; 12.9%). One patient discontinued the study; 2 patients experienced SAEs (unrelated to StrataGraft) leading to death. PRA and HLA allele reactivity was ≤ 25% at Month 3, with no persistent allogeneic DNA from StrataGraft. ConclusionsStrataGraft was well tolerated by patients, with a safety profile similar to autograft. StrataGraft may offer a safe alternative to autograft for DPT burns.