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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 -
Current Drug Delivery Jun 2024The regeneration of tissue damage involves a series of molecular and cellular events that can be mediated by various natural compounds. Recent studies have highlighted...
BACKGROUND
The regeneration of tissue damage involves a series of molecular and cellular events that can be mediated by various natural compounds. Recent studies have highlighted the anti-inflammatory, anti-ulcer, and skin-protecting properties of Cydonia oblonga (Quince), which are mainly attributed to phenolic compounds. These compounds may have some drawbacks when targeting wound applications, including low bioavailability at the wound site. Moreover, to overcome these limitations, surfactant-based nanovesicular systems have been developed as carriers of such compounds for wound healing.
OBJECTIVE
This study aimed to highlight the possible therapeutic potential of niosome-based hydrogel from Quince extract to stabilize and deliver the related bioactive compounds to full-thickness wounds in rats.
METHODS
The niosomal hydrogel was prepared using a thin-film hydration method with the fruit extract (70% methanol). The formulation was optimized by evaluating size, zeta potential, dispersion index, and drug encapsulation efficiency. Full-thickness wounds were created on the dorsal cervical area of Wistar rats, and histopathological analysis of biopsy specimens was conducted on the 12th day of treatment.
RESULTS
Under the study conditions, niosomal hydrogel displayed good physicochemical stability. Histopathological findings demonstrated that niosomal gel promoted angiogenesis, fibroblast maturation, collagen deposition, keratinization, and epidermal layer formation more effectively than control and hydrogel base. Furthermore, niosomal gel treatment markedly reduced inflammation. The total phenol concentration was determined to be 13.34 ± 0.90 mg gallic acid equivalents per gram of dried extract.
CONCLUSION
The niosomal hydrogel containing C. oblonga extract shows potential as a novel approach for wound healing, warranting further investigation in this field.
PubMed: 38840405
DOI: 10.2174/0115672018282735240528072715 -
Biomacromolecules Jun 2024Processing cellulose from 4-methyl morpholine -oxide (NMMO)-water solutions is a completely circular route that produces biodegradable cellulose fibers or films while...
Processing cellulose from 4-methyl morpholine -oxide (NMMO)-water solutions is a completely circular route that produces biodegradable cellulose fibers or films while recovering reusable NMMO [Guo, Y.; Cai, J.; Sun, T.; Xing, L.; Cheng, C.; Chi, K.; Xu, J.; Li, T. The purification process and side reactions in the N-methylmorpholine-N-oxide (NMMO) recovery system. , (12), 7609-7617]. Despite proven success in two-dimensional applications, challenges in transitioning to three-dimensional objects arise from the critical changes that cellulose undergoes during deposition, regeneration, and postregeneration stages. While emphasizing the critical diffusion-driven precipitation during regeneration, this investigation explores the influence of extrusion temperature, printing alignment, regeneration, and drying processes on interfilament fusion, bonding, shape integrity, and mechanical properties. Three distinct drying processes: ambient, vacuum, and freeze-drying were investigated. Tensile and flexural bending tests provided insight into the delamination of dried specimens. Ambient and vacuum drying enhanced the properties of specimens, while freeze-drying resulted in a more stable shape. The findings contribute to advancing the understanding of 3D-printing cellulose from NMMO solutions, addressing crucial aspects of the extrusion, regeneration, and drying stages for enhanced applications in sustainable manufacturing.
PubMed: 38832755
DOI: 10.1021/acs.biomac.4c00171 -
Surgical and Radiologic Anatomy : SRA May 2024We enjoyed reading the Brumpt et al. paper, which showed that a three-dimensional printed model (3DPM) of the ear was more effective than conventional cadaveric models...
We enjoyed reading the Brumpt et al. paper, which showed that a three-dimensional printed model (3DPM) of the ear was more effective than conventional cadaveric models for teaching anatomy. We would like to comment on the findings of this exciting study. In this case, the 3DPM of the ear was compared with dried bone models but not with a cadaveric specimen (with all adjacent soft tissues). The better results after the first test of students who used the 3DPMs were probably attributed to the optimized 3D representation of the ear anatomy. Also, the educational outcomes will likely be better if a more complex 3DPM is used, as it permits better visualization of the structures compared to the dried bone specimens. We certainly agree that 3DPMs have a remarkable ability to represent anatomy. Still, their effectiveness has not been proven superior to cadaveric specimens teaching complex anatomy. In conclusion, although we agree that 3DPMs have a high educational potential and can contribute to complex anatomy teaching, those models were not proven significantly more effective than cadaveric specimens in the Brumpt et al. study. The better effectiveness of 3DPMs compared to dried bone specimens (at the first test) does not mean those models are superior to specimens with retained soft tissues. Such cadaveric specimens permit visualization of complex structures and have proven valuable for teaching complex anatomy. Currently, the literature does not support the educational superiority of 3DPMs to those cadaveric specimens.
PubMed: 38819486
DOI: 10.1007/s00276-024-03398-7 -
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 -
Journal of Separation Science May 2024Determination of proteins from dried matrix spots using MS is an expanding research area. Mainly, the collected dried matrix sample is whole blood from a finger or heal... (Review)
Review
Determination of proteins from dried matrix spots using MS is an expanding research area. Mainly, the collected dried matrix sample is whole blood from a finger or heal prick, resulting in dried blood spots. However as other matrices such as plasma, serum, urine, and tear fluid also can be collected in this way, the term dried matrix spot is used as an overarching term. In this review, the focus is on advancements in the field made from 2017 up to 2023. In the first part reviews concerning the subject are discussed. After this, advancements made for clinical purposes are highlighted. Both targeted protein analyses, with and without the use of affinity extractions, as well as untargeted, global proteomic approaches are discussed. In the last part, both methodological advancements are being reviewed as well as the possibility to integrate sample preparation steps during the sample handling. The focus, of this so-called smart sampling, is on the incorporation of cell separation, proteolysis, and antibody-based affinity capture.
Topics: Humans; Chromatography, Liquid; Dried Blood Spot Testing; Proteins; Mass Spectrometry; Proteomics; Specimen Handling; Liquid Chromatography-Mass Spectrometry
PubMed: 38726749
DOI: 10.1002/jssc.202400061