Purpose
Malignancy prediction in indeterminate thyroid nodules is still challenging. We prospectively evaluated whether the combination of ultrasound (US) risk stratification and molecular testing ...improves the assessment of malignancy risk in Bethesda Category IV thyroid nodules.
Methods
Ninety-one consecutively diagnosed Bethesda Category IV thyroid nodules were prospectively evaluated before surgery by both ACR- and EU-TIRADS US risk-stratification systems and by a further US-guided fine-needle aspiration cytology (FNAC) for the following molecular testing: BRAFV600E, N-RAS codons 12/13, N-RAS codon 61, H-RAS codons 12/13, H-RAS codon 61, K-RAS codons 12/13, and K-RAS codon 61 point-mutations, as well as PAX8/PPARγ, RET/PC1, and RET/PTC 3 rearrangements.
Results
At histology, 37% of nodules were malignant. No significant association was found between malignancy and either EU- or ACR-TIRADS. In total, 58 somatic mutations were identified, including 3 BRAFV600E (5%), 5 N-RAS 12/13 (9%), 13 N-RAS 61 (22%), 7 H-RAS 12/13 (12%), 11 H-RAS 61 (19%), 6 K-RAS 12/13 (10%), 8 K-RAS 61 (14%) mutations and 2 RET/PTC1 (4%), 0 RET/PTC 3 (0%), 3 PAX8/PPARγ (5%) rearrangements. At least one somatic mutation was found in 28% and 44% of benign and malignant nodules, respectively, although malignancy was not statistically associated with the outcome of the mutational test. However, the combination of ACR-, but not EU-, TIRADS with the presence of at least one somatic mutation, was significantly associated with malignant histology (
P
= 0.03).
Conclusion
US risk stratification and FNAC molecular testing may synergistically contribute to improve malignancy risk estimate of Bethesda category IV thyroid nodules.
Objectives:
To investigate the extent of synaptic loss, and the contribution of gray matter (GM) inflammation and demyelination to synaptic loss, in multiple sclerosis (MS) brain tissue.
Methods:
...This study was performed on two different post-mortem series of MS and control brains, including deep GM and cortical GM. MS brain samples had been specifically selected for the presence of active demyelinating GM lesions. Over 1,000,000 individual synapses were identified and counted using confocal microscopy, and further characterized as glutamatergic/GABAergic. Synaptic counts were also correlated with neuronal/axonal loss.
Results:
Important synaptic loss was observed in active demyelinating GM lesions (−58.9%), while in chronic inactive GM lesions, synaptic density was only mildly reduced compared to adjacent non-lesional gray matter (NLGM) (−12.6%). Synaptic loss equally affected glutamatergic and GABAergic synapses. Diffuse synaptic loss was observed in MS NLGM compared to control GM (−21.2% overall).
Conclusion:
This study provides evidence, in MS brain tissue, of acute synaptic damage/loss during active GM inflammatory demyelination and of synaptic reorganization in chronically demyelinated GM, affecting equally glutamatergic and GABAergic synapses. Furthermore, this study provides a strong indication of widespread synaptic loss in MS NLGM also independently from focal GM demyelination.
Chickpea is the third most important grain legume in the world after common bean and pea. Ascochyta blight (AB) of chickpea, caused by
Ascochyta rabiei
, is the most damaging disease of this crop ...worldwide. Losses may reach 100%, with damage affecting yield and seed quality. AB produces lesions to all aerial plant tissues: leaves, petioles, flowers and pods. The causal agent can survive on or in the seeds and up to four years in stubble. Genetic resistance is the preferred tool for the management of this complex disease. This is the most effective and inexpensive way of controlling biotic stresses and the major goal of many breeding programs. In Argentina, where AB was first detected in 2011, the most widely used commercial varieties, Chañaritos S-156, Norteño, Felipe UNC-INTA and Kiara UNC-INTA, are susceptible to
A. rabiei
. In this work, 109 genotypes carrying multiple resistance to different pathogens were evaluated against local
A. rabiei
under controlled conditions. The results showed that all genotypes were affected by this fungus, without occurrence of asymptomatic plants. The genotypes ranged from resistant to highly susceptible. According to disease reaction, the genotypes were classified as: resistant (2.75%),moderately resistant (32.09%), susceptible (60.55%) and highly susceptible (1.83%). This is the first research developed in Argentina to identify resistant genotypes and sources of resistance that contribute to breeding programs.
A newly proposed form of brain structural plasticity consists of non-newly generated, "immature" neurons of the adult cerebral cortex. Similar to newly generated neurons, these cells express the ...cytoskeletal protein Doublecortin (DCX), yet they are generated prenatally and then remain in a state of immaturity for long periods. In rodents, the immature neurons are restricted to the paleocortex, whereas in other mammals, they are also found in neocortex. Here, we analyzed the DCX-expressing cells in the whole sheep brain of both sexes to search for an indicator of structural plasticity at a cellular level in a relatively large-brained, long-living mammal. Brains from adult and newborn sheep (injected with BrdU and analyzed at different survival times) were processed for DCX, cell proliferation markers (Ki-67, BrdU), pallial/subpallial developmental origin (
,
), and neuronal/glial antigens for phenotype characterization. We found immature-like neurons in the whole sheep cortex and in large populations of DCX-expressing cells within the external capsule and the surrounding gray matter (claustrum and amygdala). BrdU and Ki-67 detection at neonatal and adult ages showed that all of these DCX
cells were generated during embryogenesis, not after birth. These results show that the adult sheep, unlike rodents, is largely endowed with non-newly generated neurons retaining immature features, suggesting that such plasticity might be particularly important in large-brained, long-living mammals.
Brain plasticity is important in adaptation and brain repair. Structural changes span from synaptic plasticity to adult neurogenesis, the latter being highly reduced in large-brained, long-living mammals (e.g., humans). The cerebral cortex contains "immature" neurons, which are generated prenatally and then remain in an undifferentiated state for long periods, being detectable with markers of immaturity. We studied the distribution and developmental origin of these cells in the whole brain of sheep, relatively large-brained, long-living mammals. In addition to the expected cortical location, we also found populations of non-newly generated neurons in several subcortical regions (external capsule, claustrum, and amygdala). These results suggests that non-neurogenic, parenchymal structural plasticity might be more important in large mammals with respect to adult neurogenesis.
Amyotrophic lateral sclerosis (ALS) is a complex disease characterized by the interplay of genetic and environmental factors for which, despite decades of intense research, diagnosis remains rather ...delayed, and most therapeutic options fail. Therefore, unravelling other potential pathogenetic mechanisms and searching for reliable markers are high priorities. In the present study, we employ the SOMAscan assay, an aptamer-based proteomic technology, to determine the circulating proteomic profile of ALS patients. The expression levels of ~1300 proteins were assessed in plasma, and 42 proteins with statistically significant differential expression between ALS patients and healthy controls were identified. Among these, four were upregulated proteins, Thymus- and activation-regulated chemokine, metalloproteinase inhibitor 3 and nidogen 1 and 2 were selected and validated by enzyme-linked immunosorbent assays in an overlapping cohort of patients. Following statistical analyses, different expression patterns of these proteins were observed in the familial and sporadic ALS patients. The proteins identified in this study might provide insight into ALS pathogenesis and represent potential candidates to develop novel targeted therapies.
Since 2005, two cases of natural bovine spongiform encephalopathies (BSE) have been reported in goats. Furthermore, experimental transmissions of classical (C-BSE) and atypical (L-BSE) forms of BSE ...in goats were also reported. To minimize further spreading of prion diseases in small ruminants the development of a highly sensitive and specific test for ante-mortem detection of infected animals would be of great value. Recent studies reported high diagnostic value of a second generation of cerebrospinal fluid (CSF) Real-Time Quaking-Induced Conversion (RT-QuIC) assay across a wide spectrum of human prions. Here, we applied this improved QuIC (IQ-CSF) for highly efficient detection of TSEs prion protein in goat cerebrospinal fluid. IQ-CSF sensitivity and specificity were evaluated on CSF samples collected at disease endpoint from goats naturally and experimentally infected with scrapie or bovine isolates of C-BSE and L-BSE, respectively. Next, CSF samples collected from L-BSE infected goats during pre-symptomatic stage were also analysed. PrP
associated seeding activity was detected at early time points after experimental inoculation, with an average time of 439 days before clinical symptoms appeared. Taken together these data are indicative of the great potential of this in vitro prion amplification assay as ante-mortem TSE test for live and asymptomatic small ruminants.
Ascochyta blight is the major disease affecting chickpea (Cicer arietinum) around the world. Since the first report of Ascochyta rabiei's isolation in Argentina in 2012, the pathogen has caused ...severe economic losses in crop production; so, the detection and rapid identification of the pathogen in early stages is key for the management of the disease. In this work, a traditional PCR procedure for detection of A. rabiei directly from plant tissues has been described based on beta-tubulin gene. The TP-6/TP-9 specific primers designed, amplified only a single PCR band of 770 bp from A. rabiei. The specificity of the primers was checked using 12 isolates of A. rabiei and DNA from 10 other different fungi including common pathogens of chickpea as Alternaria alternata, Botrytis cinerea, Sclerotinia sclerotiorum and Phoma medicaginis that cause similar symptoms. The detection sensitivity with primers was 2 × 104 ng μl−1 genomic DNA. In inoculated plant material, PCR amplification gave a band of the expected size and no amplification was observed when DNA was from healthy and uninoculated plants. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods. The PCR-based method developed here can simplify both plant disease diagnosis, and pathogen monitoring in an early phase, as well as aid in effective management practices that avoid the disease advance and minimize losses.
•A rapid, sensitive, and effective molecular method to detect A. rabiei in early stages of the disease was developed.•A. rabiei specific primers were designed from beta-tubulin gene.•Field samples with incipient symptoms of Ascochyta bligth of 50 sites were monitored using the molecular method developed.
Apple valsa canker, caused by the fungus
Valsa ceratosperma
, is one of the most destructive diseases of this crop. Conventional fungicide treatments are not effective enough; therefore, it is ...necessary to develop alternatives for the control of this pathogen. The use of
Trichoderma
strains as a biocontrol agent is a promising strategy for the environmentally friendly management of this disease. In this study, seven different isolates of
Trichoderma
spp. were obtained from the rhizospheric soil of healthy apple plants. The antagonistic capacity of the isolates against the pathogen
V. ceratosperma
was evaluated using the dual culture method and by the production of antibiotic non-volatile compounds. In the dual culture method, the interaction zone was observed by differential interference contrast microscopy and confocal laser scanning microscopy. The isolates RN-13, RN-33, and RN-34 inhibited more than 75% of mycelial growth, whereas RN-19, RN-16, and RN-15 inhibited growth by 65–72% with respect to the control. The mycoparasitic activity was evidenced by the coiling of hyphae around pathogen hyphae, appressorium-like structures, morphological deformations, and disintegration of mycelial walls. Apple shoots inoculated with
Trichoderma
isolates and then infected with
V. ceratosperma
showed the antagonist capacity of RN-16, RN-19, RN-15, and RN-18, which produced a significant decrease in the size of the lesion. Overall, the results showed the great potential of
Trichoderma
as a biocontrol agent against apple tree valsa canker. Further studies of this fungal interaction are required to design effective control strategies for the protection of apple tree orchards from this disease.
Leigh syndrome (LS) associated with cytochrome c oxidase (COX) deficiency is an early onset, fatal mitochondrial encephalopathy, leading to multiple neurological failure and eventually death, usually ...in the first decade of life. Mutations in SURF1, a nuclear gene encoding a mitochondrial protein involved in COX assembly, are among the most common causes of LS. LSSURF1 patients display severe, isolated COX deficiency in all tissues, including cultured fibroblasts and skeletal muscle. Recombinant, constitutive SURF1−/− mice show diffuse COX deficiency, but fail to recapitulate the severity of the human clinical phenotype. Pigs are an attractive alternative model for human diseases, because of their size, as well as metabolic, physiological and genetic similarity to humans. Here, we determined the complete sequence of the swine SURF1 gene, disrupted it in pig primary fibroblast cell lines using both TALENs and CRISPR/Cas9 genome editing systems, before finally generating SURF1−/− and SURF1−/+ pigs by Somatic Cell Nuclear Transfer (SCNT). SURF1−/− pigs were characterized by failure to thrive, muscle weakness and highly reduced life span with elevated perinatal mortality, compared to heterozygous SURF1−/+ and wild type littermates. Surprisingly, no obvious COX deficiency was detected in SURF1−/− tissues, although histochemical analysis revealed the presence of COX deficiency in jejunum villi and total mRNA sequencing (RNAseq) showed that several COX subunit-encoding genes were significantly down-regulated in SURF1−/− skeletal muscles. In addition, neuropathological findings, indicated a delay in central nervous system development of newborn SURF1−/− piglets. Our results suggest a broader role of sSURF1 in mitochondrial bioenergetics.
•The full sequence of pig SURF1 gene was determined.•SURF1 gene was disrupted in pig by gene editing and somatic cell nuclear transfer.•SURF1−/− piglets showed an early onset lethal phenotype.•Mitochondrial bioenergetics was impaired in the skeletal muscle of SURF1−/− pigs.
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