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IMA Fungus Jun 2024Molecular studies of fungi within the order Laboulbeniales (Ascomycota, Pezizomycotina) have been hampered for years because of their minute size, inability to grow in...
Molecular studies of fungi within the order Laboulbeniales (Ascomycota, Pezizomycotina) have been hampered for years because of their minute size, inability to grow in axenic culture, and lack of reliable and cost-efficient DNA extraction protocols. In particular, the genus Laboulbenia is notorious for low success with DNA extraction and polymerase chain reaction (PCR) amplification. This is attributed to the presence of melanin, a molecule known to inhibit PCR, in the cells. We evaluated the efficacy of a standard single cell-based DNA extraction protocol by halving the recommended amount of reagents to reduce the cost per extraction and adding bovine serum albumin (BSA) during the multiple displacement amplification step to reverse the effect of melanin. A total of 196 extractions were made, 111 of which were successful. We found that halving the reagents used in the single cell-based extraction kit did not significantly affect the probability of successful DNA extraction. Using the halved protocol reduces cost and resource consumption. Moreover, there was no significant difference in the probability of successfully extracting DNA based on whether BSA was added or not, suggesting that the amount of melanin present in cells of the thallus has no major inhibitory effect on PCR. We generated 277 sequences from five loci, but amplification of the internal transcribed spacer region, the mitochondrial small subunit rDNA, and protein-coding genes remains challenging. The probability of successfully extracting DNA from Laboulbeniales was also impacted by specimen storage methods, with material preserved in > 95% ethanol yielding higher success rates compared to material stored in 70% ethanol and dried material. We emphasize the importance of proper preservation of material and propose the design of Laboulbeniales-specific primers to overcome the problems of primer mismatches and contaminants. Our new insights apply not only to the genus Laboulbenia; Laboulbeniales generally are understudied, and the vast majority of species remain unsequenced. New and approachable molecular developments will benefit the study of Laboulbeniales, helping to elucidate the true diversity and evolutionary relationships of these peculiar microfungi.
PubMed: 38863065
DOI: 10.1186/s43008-024-00146-9 -
Nature Communications Jun 2024The SpaceX Inspiration4 mission provided a unique opportunity to study the impact of spaceflight on the human body. Biospecimen samples were collected from four crew...
The SpaceX Inspiration4 mission provided a unique opportunity to study the impact of spaceflight on the human body. Biospecimen samples were collected from four crew members longitudinally before (Launch: L-92, L-44, L-3 days), during (Flight Day: FD1, FD2, FD3), and after (Return: R + 1, R + 45, R + 82, R + 194 days) spaceflight, spanning a total of 289 days across 2021-2022. The collection process included venous whole blood, capillary dried blood spot cards, saliva, urine, stool, body swabs, capsule swabs, SpaceX Dragon capsule HEPA filter, and skin biopsies. Venous whole blood was further processed to obtain aliquots of serum, plasma, extracellular vesicles and particles, and peripheral blood mononuclear cells. In total, 2,911 sample aliquots were shipped to our central lab at Weill Cornell Medicine for downstream assays and biobanking. This paper provides an overview of the extensive biospecimen collection and highlights their processing procedures and long-term biobanking techniques, facilitating future molecular tests and evaluations.As such, this study details a robust framework for obtaining and preserving high-quality human, microbial, and environmental samples for aerospace medicine in the Space Omics and Medical Atlas (SOMA) initiative, which can aid future human spaceflight and space biology experiments.
Topics: Humans; Space Flight; Biological Specimen Banks; Specimen Handling; Astronauts
PubMed: 38862509
DOI: 10.1038/s41467-024-48806-z -
F1000Research 2023The sternum exhibits unique anatomical variations with major clinical and forensic implications. This study is devoted to providing baseline epidemiological information... (Observational Study)
Observational Study
The sternum exhibits unique anatomical variations with major clinical and forensic implications. This study is devoted to providing baseline epidemiological information about the sternal foramen and variant xiphoid morphology in Ethiopia. Two extremely interesting and unusual variations of the sternal foramen are also discussed. This observational study was carried out using dried adult human sternum obtained from skeletal remains samples brought for medicolegal examination over a period of 4 years. A total of 94 dried adult human sternums (66 males (70.2%) and 28 females (29.8%)) were obtained with an age range of 21 to 57 years and a mean age at death of 38.383 ± 11.3480 years. Dried human sternum specimens were morphologically examined, and morphometric parameters were recorded and photographed. A sternal foramen was found in 18 specimens (19.1%); 17 were male and one was female. A single sternal foramen was observed in 83.3% (n=15/18) of the sternal bodies and 11.1% (n=2/18) of the xiphoid processes (both males). In addition, a double sternal foramen was observed in a single male specimen on the mesosternum and xiphoid process. The most common sternal foramen site was at the fifth costochondral junction level. The xiphoid process was present in 77 samples and ended as a single process in 83.1% (n=64/77) of samples. In 15.6% (n=12/77) of the samples, the xiphoid process was bifurcated and trifurcated in a single male (1.3%) specimen. The sternal foramen and variation in xiphoid morphology are common anatomical variations in Ethiopia. The findings of the current study highlight the necessity of strict precautionary measures during sternal procedures in this study population. In addition, such incidental findings during radiologic and autopsy procedures should be properly evaluated to avoid misdiagnosis and misinterpretation of such findings as traumatic or pathologic conditions.
Topics: Humans; Sternum; Male; Female; Ethiopia; Adult; Middle Aged; Young Adult; Anatomic Variation; Xiphoid Bone
PubMed: 38845617
DOI: 10.12688/f1000research.133151.1 -
Plant Disease May 2024Since the emergence of Ug99 wheat stem rust in Uganda in 1998 (Pretorius et al. 2000), the threat of movement into South Asia has been a concern due to long-distance...
Since the emergence of Ug99 wheat stem rust in Uganda in 1998 (Pretorius et al. 2000), the threat of movement into South Asia has been a concern due to long-distance dispersal capacity of airborne spores (Brown and Hovmøller 2002; Singh et al. 2008; Meyer et al. 2017). Increased preparedness by comprehensive rust surveillance efforts and development and deployment of resistant cultivars in advance of an incursion into South Asia has been one of the success stories of the Borlaug Global Rust Initiative (Sharma et al. 2013). In November 2023, an off-season rust survey was conducted in Marpha, Gandaki and Bagmati provinces in Nepal. Rust was only observed at two sites, Dangdunge of Dolakha district and Mude of Sindhupalchok district, where spring wheat was grown as fodder crop outside the main cropping season. Rust infected wheat leaves (10-15 leaves per site) were air dried and sealed in envelopes that were shipped under permit to the Global Rust Reference Center, Denmark. Bulk samples of stem rust, Puccinia graminis f.sp. tritici (Pgt), were recovered from both envelopes, and single pustule isolates were raised and multiplied on Morocco and McNair. Meanwhile, specimens of dry leaves were subjected to SSR genotyping according to standard procedures (Patpour et al. 2022). One distinct multi-locus Pgt genotype was observed, identical to and representing 99% of Ug99 isolates within Clade I collected in East Africa between 2012-2022. A Pgt single pustule isolate from each of the sampling sites were inoculated onto 20 internationally agreed stem rust differential lines using standard procedures, and 14 supplementary lines providing additional resolution of pathogen virulence (Patpour et al. 2022). The pathotyping was repeated in two independent experiments, which resulted in the infection type pattern of Pgt race TTKTT (Supplementary Table 1). Additional independent SSR genotype assays of recovered isolates confirmed the prevalent genotype of Clade I (Patpour et al. 2022; Szabo et al. 2022). This first detection of Ug99 race TTKTT in South Asia emphasizes the need for continued coordinated international surveillance efforts and utilization of diverse sources of resistance to control stem rust in wheat. New surveillance efforts in Nepal during February-March 2024 did not reveal additional cases of wheat stem rust. However, more detailed and sustained rust surveillance efforts, assessment of the vulnerability of current wheat crops to Ug99 and other races of stem-, stripe/yellow- and leaf rust, as well as intensified breeding for rust resistance throughout the region is strongly recommended to meet current and future plant health risks.
PubMed: 38812370
DOI: 10.1094/PDIS-03-24-0644-PDN -
Bioengineering (Basel, Switzerland) May 2024The application of calcium coacervates (CCs) may hold promise for dental hard tissue remineralization. The aim of this study was to evaluate the effect of the...
The application of calcium coacervates (CCs) may hold promise for dental hard tissue remineralization. The aim of this study was to evaluate the effect of the infiltration of artificial enamel lesions with a CC and its single components including polyacrylic acid (PAA) compared to that of the self-assembling peptide P-4 in a pH-cycling (pHC) model. Enamel specimens were prepared from bovine incisors, partly varnished, and stored in demineralizing solution (DS; pH 4.95; 17 d) to create two enamel lesions per sample. The specimens were randomly allocated to six groups (n = 15). While one lesion per specimen served as the no-treatment control (NTC), another lesion (treatment, T) was etched (HPO, 5 s), air-dried and subsequently infiltrated for 10 min with either a CC (10 mg/mL PAA, 50 mM CaCl (Ca) and 1 M KHPO (PO)) (groups CC and CC + DS) or its components PAA, Ca or PO. As a commercial control, the self-assembling peptide P-4 (Curodont Repair, Credentis, Switzerland) was tested. The specimens were cut perpendicularly to the lesions, with half serving as the baseline (BL) while the other half was exposed to either a demineralization solution for 20 d (pH 4.95; group CC + DS) or pHC for 28 d (pH 4.95, 3 h; pH 7, 21 h; all five of the other groups). The difference in integrated mineral loss between the lesions at BL and after the DS or pHC, respectively, was analyzed using transversal microradiography (ΔΔZ = ΔZ - ΔZ). Compared to the NTC, the mineral gain in the T group was significantly higher in the CC + DS, CC and PAA ( < 0.05, Wilcoxon). In all of the other groups, no significant differences between treated and untreated lesions were detected ( > 0.05). Infiltration with the CC and PAA resulted in a consistent mineral gain throughout the lesion body. The CC as well as its component PAA alone promoted the remineralization of artificial caries lesions in the tested pHC model. Infiltration with PAA further resulted in mineral gain in deeper areas of the lesion body.
PubMed: 38790332
DOI: 10.3390/bioengineering11050465 -
ENeurologicalSci Jun 2024A case-control study of sporadic amyotrophic lateral sclerosis (ALS) in a mountainous village in the French Alps discovered an association of cases with a history of...
A case-control study of sporadic amyotrophic lateral sclerosis (ALS) in a mountainous village in the French Alps discovered an association of cases with a history of eating wild fungi (false morels) collected locally and initially identified and erroneously reported as . Specialist re-examination of dried specimens of the ALS-associated fungi demonstrated they were members of the group, namely , species that have been reported to contain substantially higher concentrations of gyromitrin than present in . Gyromitrin is metabolized to monomethylhydrazine, which is responsible not only for the acute oral toxic and neurotoxic properties of false morels but also has genotoxic potential with proposed mechanistic relevance to the etiology of neurodegenerative disease. Most ALS patients had a slow- or intermediate-acetylator phenotype predicted by - () genotyping, which would increase the risk for neurotoxic and genotoxic effects of gyromitrin metabolites.
PubMed: 38770222
DOI: 10.1016/j.ensci.2024.100502 -
Cureus May 2024Introduction The aim of this study was to evaluate the activity of an ethanolic extract of Aloe vera on and Materials and methods A total of 42 heat-cured acrylic...
Introduction The aim of this study was to evaluate the activity of an ethanolic extract of Aloe vera on and Materials and methods A total of 42 heat-cured acrylic resin specimens were made and divided into three groups according to the disinfection method: (1) Corega disinfectant tablets; (2) ethanol extract of Aloe vera; and (3) distilled water (as a control group). Fresh Aloe vera whole leaves were washed with distilled water, chopped into small pieces, air-dried, and ground into powder. The powder was extracted with 95% ethanol. The acrylic specimens were contaminated with and , and then the specimens were immersed in study solutions for three minutes. The viable colonies were counted using the colony-forming units (CFU) method. Results The results showed a decrease in the number of CFU for denture tablets and Aloe vera ethanoic extract groups compared to the negative control group. There were no significant statistical differences between the denture tablet group and the Aloe vera ethanolic extract group (P < 0.05). Aloe vera ethanolic extract groups significantly decreased the number of CFU compared to the negative control group and less compared to the denture tablet, where significant statistical differences were found between the tablet group and the Aloe vera ethanolic extract group. Conclusions Within the limitations of this study, it was concluded that Aloe vera extract was effective against and when acrylic resin specimens were immersed for three minutes.
PubMed: 38726356
DOI: 10.7759/cureus.59916 -
The Lancet. Infectious Diseases May 2024The first licensed malaria vaccine, RTS,S/AS01, confers moderate protection against symptomatic disease. Because many malaria infections are asymptomatic, we conducted a...
Genotypic analysis of RTS,S/AS01 malaria vaccine efficacy against parasite infection as a function of dosage regimen and baseline malaria infection status in children aged 5-17 months in Ghana and Kenya: a longitudinal phase 2b randomised controlled trial.
BACKGROUND
The first licensed malaria vaccine, RTS,S/AS01, confers moderate protection against symptomatic disease. Because many malaria infections are asymptomatic, we conducted a large-scale longitudinal parasite genotyping study of samples from a clinical trial exploring how vaccine dosing regimen affects vaccine efficacy.
METHODS
Between Sept 28, 2017, and Sept 25, 2018, 1500 children aged 5-17 months were randomly assigned (1:1:1:1:1) to receive four different RTS,S/AS01 regimens or a rabies control vaccine in a phase 2b open-label clinical trial in Ghana and Kenya. Participants in the four RTS,S groups received two full doses at month 0 and month 1 and either full doses at month 2 and month 20 (group R012-20); full doses at month 2, month 14, month 26, and month 38 (group R012-14); fractional doses at month 2, month 14, month 26, and month 38 (group Fx012-14; early fourth dose); or fractional doses at month 7, month 20, and month 32 (group Fx017-20; delayed third dose). We evaluated the time to the first new genotypically detected infection and the total number of new infections during two follow-up periods (12 months and 20 months) in more than 36 000 dried blood spot specimens from 1500 participants. To study vaccine effects on time to the first new infection, we defined vaccine efficacy as one minus the hazard ratio (HR; RTS,S vs control) of the first new infection. We performed a post-hoc analysis of vaccine efficacy based on malaria infection status at first vaccination and force of infection by month 2. This trial (MAL-095) is registered with ClinicalTrials.gov, NCT03281291.
FINDINGS
We observed significant and similar vaccine efficacy (25-43%; 95% CI union 9-53) against first new infection for all four RTS,S/AS01 regimens across both follow-up periods (12 months and 20 months). Each RTS,S/AS01 regimen significantly reduced the mean number of new infections in the 20-month follow-up period by 1·1-1·6 infections (95% CI union 0·6-2·1). Vaccine efficacy against first new infection was significantly higher in participants who were infected with malaria (68%; 95% CI 50-80) than in those who were uninfected (37%; 23-48) at the first vaccination (p=0·0053).
INTERPRETATION
All tested dosing regimens blocked some infections to a similar degree. Improved vaccine efficacy in participants infected during vaccination could suggest new strategies for highly efficacious malaria vaccine development and implementation.
FUNDING
GlaxoSmithKline Biologicals SA, PATH, Bill & Melinda Gates Foundation, and the German Federal Ministry of Education and Research.
PubMed: 38723650
DOI: 10.1016/S1473-3099(24)00179-8 -
Gels (Basel, Switzerland) Mar 2024Tooth avulsion and delayed replantation may cause inflammatory responses and root resorption of the tooth. The aim of this study is to investigate the effect of a...
BACKGROUND/AIM
Tooth avulsion and delayed replantation may cause inflammatory responses and root resorption of the tooth. The aim of this study is to investigate the effect of a doxycycline-loaded nitric oxide-releasing nanomatrix (DN) gel on the delayed replantation of avulsed rat teeth, with a focus on assessing the gel's potential to promote regeneration and inhibit complications associated with the replantation process.
MATERIALS AND METHODS
Twenty-four right maxillary first molars from male Sprague-Dawley rats were atraumatically extracted using sterile extraction forceps. The molars were dried for 1 h at room temperature (approximately 23 °C) and divided into four groups according to the root conditioning methods after extra-alveolar 60-min drying: Group 1, no root conditioning treatment prior to replantation; Group 2, soaking in 2% NaF solution for 5 min before replantation; Group 3, 5-min soaking in NO gel and injection of the gel into the alveolar socket; Group 4, 5-min soaking in DN gel and injection of the gel into the alveolar socket before replantation. The animals were euthanized four weeks after the operation and the specimens were evaluated histologically.
RESULTS
The use of NO gel alone showed better anti-inflammatory and periodontal effects than the control group, but it did not show a significant effect compared to the group using NaF. When using NO gel loaded with doxycycline, it showed a significant anti-inflammatory effect compared to the control group and showed a similar inhibitory effect to the group using NaF.
CONCLUSIONS
Within the limits of this study, in delayed replantation situations, the control of inflammatory resorption and replacement resorption is an important factor for achieving a better prognosis of replanted teeth. Root surface treatment with DN gel decreased root resorption after delayed replantation.
PubMed: 38667632
DOI: 10.3390/gels10040213 -
Environment International May 2024Use of capillary blood devices for exposome research can deepen our understanding of the intricate relationship between environment and health, and open up new avenues... (Comparative Study)
Comparative Study
Use of capillary blood devices for exposome research can deepen our understanding of the intricate relationship between environment and health, and open up new avenues for preventive and personalized medicine, particularly for vulnerable populations. While the potential of these whole blood devices to accurately measure chemicals and metabolites has been demonstrated, how untargeted metabolomics data from these samplers can be integrated with previous and ongoing environmental health studies that have used conventional blood collection approaches is not yet clear. Therefore, we performed a comprehensive comparison between relative-quantitative metabolite profiles measured in venous blood collected with dried whole blood microsamplers (DBM), dried whole blood spots (DBS), and plasma from 54 mothers in an ethnically diverse population. We determined that a majority of the 309 chemicals and metabolites showed similar median intensity rank, moderate correlation, and moderate agreement between participant-quantiled intraclass correlation coefficients (ICCs) for pair-wise comparisons among the three biomatrices. In particular, whole blood sample types, DBM and DBS, were in highest agreement across metabolite comparison metrics, followed by metabolites measured in DBM and plasma, and then metabolites measured in DBS and plasma. We provide descriptive characteristics and measurement summaries as a reference database. This includes unique metabolites that were particularly concordant or discordant in pairwise comparisons. Our results demonstrate that the range of metabolites from untargeted metabolomics data collected with DBM, DBS, and plasma provides biologically relevant information for use in independent exposome investigations. However, before meta-analysis with combined datasets are performed, robust statistical approaches that integrate untargeted metabolomics data collected on different blood matrices need to be developed.
Topics: Humans; Metabolomics; Female; Dried Blood Spot Testing; Environmental Health; Adult; Plasma; Blood Specimen Collection; Pregnancy; Exposome
PubMed: 38657407
DOI: 10.1016/j.envint.2024.108663