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Pathogens ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 106
Author(s):  
Marcos Rogério André ◽  
Ana Cláudia Calchi ◽  
Maria Eduarda Chiaradia Furquim ◽  
Isabela de Andrade ◽  
Paulo Vitor Cadina Arantes ◽  
...  

Even though the epidemiology of tick-borne agents (TBA) in dogs has been extensively investigated around the world, the occurrence, vectors involved, and molecular identity of these agents in cats remains elusive in many regions. Among TBA, Ehrlichia, Anaplasma, Babesia, Cytauxzoon, and Hepatozoon are responsible for diseases with non-specific clinical signs in cats, making essential the use of molecular techniques for accurate diagnosis and proper treatment. The present work aimed to investigate the occurrence and molecular identity of tick-borne agents (Ehrlichia, Anaplasma, Babesia/Theileria, Cytauxzoon, and Hepatozoon) in cats from southeastern (states of São Paulo (SP) and Minas Gerais (MG)) and northern (state of Rondônia (RO)) Brazil. For this purpose, 390 blood samples were collected from domiciled cats in MG (n = 155), SP (n = 151), and RO(n = 84) states, submitted to DNA extraction and PCR assays for Ehrlichia spp. (dsb gene), Anaplasma spp. (rrs gene), piroplasmids (18S rRNA gene), and Hepatozoon spp. (18S rRNA gene), sequencing, and phylogenetic inferences. The overall positivity for Anaplasma spp., Ehrlichia spp., Babesia/Theileria spp., Cytauxzoon spp., and Hepatozoon spp. were 7.4% (12.3% (MG) and 6.6% (SP)), 2% (4.5% (MG) and 0.6% (SP)), 0.7% (0.6% (MG), 0.6% (SP) and 1.2% (RO)), 27.2% (41.9% (MG), 24.5% (SP) and 4.8% (RO), and 0%, respectively. The phylogenetic analysis grouped the obtained sequences with ‘Candidatus Anaplasma amazonensis’, A. platys, B. vogeli, and Cytauxzoon sp. previously detected in wild felids from Brazil. qPCR specific for E. canis based on the dsb gene confirmed the molecular identity of the detected ehrlichial agent. The present study expanded the list and geographical distribution of hemoparasites in cats. ‘Candidatus Anaplasma amazonensis’, recently detected in sloths from northern Brazil, was described for the first time in cats. This is the first report of piroplasmids infecting cats in northern Brazil. Coinfection by Cytauxzoon and other TBA (Ehrlichia, Anaplasma, and B. vogeli) reported in the present study raises the need for veterinary practitioners’ awareness of cats parasitized by multiple TBA.


ASN NEURO ◽  
2022 ◽  
Vol 14 ◽  
pp. 175909142110733
Author(s):  
Louise Thiry ◽  
Jean-Pierre Clément ◽  
Rainer Haag ◽  
Timothy E Kennedy ◽  
Stefano Stifani

Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-derived motor neurons (MNs) have been used to study MN diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis. Modeling MN disease using hiPSC-based approaches requires culture conditions that can recapitulate in a dish the events underlying differentiation, maturation, aging, and death of MNs. Current hiPSC-derived MN-based applications are often hampered by limitations in our ability to monitor MN morphology, survival, and other functional properties over a prolonged timeframe, underscoring the need for improved long-term culture conditions. Here we describe a cytocompatible dendritic polyglycerol amine (dPGA) substrate-based method for prolonged culture of hiPSC-derived MNs. We provide evidence that MNs cultured on dPGA-coated dishes are more amenable to long-term study of cell viability, molecular identity, and spontaneous network electrophysiological activity. The present study has the potential to improve hiPSC-based studies of human MN biology and disease. We describe the use of a new coating substrate providing improved conditions for long-term cultures of human iPSC-derived motor neurons, thus allowing evaluation of cell viability, molecular identity, spontaneous network electrophysiological activity, and single-cell RNA sequencing of mature motor neurons.


Zootaxa ◽  
2021 ◽  
Vol 5082 (6) ◽  
pp. 572-582
Author(s):  
RAFAILIA ALEXIOU ◽  
GEORGIA STAMOU ◽  
STYLIANI MINOUDI ◽  
FILIO TOURLI ◽  
VALENTINA TSARTSIANIDOU ◽  
...  

For the genus Diaphanosoma Fischer (Cladocera) the species’ name “D. brachyurum” has been widely used for many decades to identify other species belonging to this genus. To clarify the diversity of the genus in Greek lakes in the present study, we morphologically and genetically identified the Diaphanosoma species occurring in eight lakes. Three hundred twenty-nine Diaphanosoma individuals were morphologically examined, while for the genetic analyses the mtDNA COI gene was sequenced in 48 individuals. Combining the morphological and genetic results, we verified the occurrence of D. mongolianum, D. orghidani and D. macedonicum in our study area. We could not confirm prior records of D. brachyurum and D. lacustris while we provide the molecular identity of D. macedonicum. Furthermore, we highlight the need to check whether the European D. mongolianum populations are characterised of mitochondrial discordance and hybridization as the individuals from the Asian type locality of the species. Our results support the importance of combining both approaches to correctly identify taxonomic species, despite the extra effort and cost during the sample analysis.  


2021 ◽  
Vol 15 ◽  
Author(s):  
Brenda Toscano Márquez ◽  
Anna A. Cook ◽  
Max Rice ◽  
Alexia Smileski ◽  
Kristen Vieira-Lomasney ◽  
...  

Patterned cell death is a common feature of many neurodegenerative diseases. In patients with autosomal-recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) and mouse models of ARSACS, it has been observed that Purkinje cells in anterior cerebellar vermis are vulnerable to degeneration while those in posterior vermis are resilient. Purkinje cells are known to express certain molecules in a highly stereotyped, patterned manner across the cerebellum. One patterned molecule is zebrin, which is expressed in distinctive stripes across the cerebellar cortex. The different zones delineated by the expression pattern of zebrin and other patterned molecules have been implicated in the patterning of Purkinje cell death, raising the question of whether they contribute to cell death in ARSACS. We found that zebrin patterning appears normal prior to disease onset in Sacs–/– mice, suggesting that zebrin-positive and -negative Purkinje cell zones develop normally. We next observed that zebrin-negative Purkinje cells in anterior lobule III were preferentially susceptible to cell death, while anterior zebrin-positive cells and posterior zebrin-negative and -positive cells remained resilient even at late disease stages. The patterning of Purkinje cell innervation to the target neurons in the cerebellar nuclei (CN) showed a similar pattern of loss: neurons in the anterior CN, where inputs are predominantly zebrin-negative, displayed a loss of Purkinje cell innervation. In contrast, neurons in the posterior CN, which is innervated by both zebrin-negative and -positive puncta, had normal innervation. These results suggest that the location and the molecular identity of Purkinje cells determine their susceptibility to cell death in ARSACS.


Plant Disease ◽  
2021 ◽  
Author(s):  
Hannah M Rivedal ◽  
Javier Felipe Tabima ◽  
Alexandra G Stone ◽  
Ken Johnson

Winter squash (Cucurbita maxima cv. ‘Golden Delicious’) produced in Oregon’s Willamette Valley for edible seed production has experienced significant yield losses due to a soilborne disease. The symptoms associated with this disease problem include root rot, crown rot and vascular discoloration in the stems leading to a severe late season wilt and plant collapse. Through field surveys, Fusarium oxysporum, F. solani, F. culmorum-like fungi, Plectosphaerella cucumerina, and Setophoma terrestris were identified to be associated with diseased tissues, and each produced symptoms of root rot, crown rot or stem discoloration in preliminary pathogenicity trials. In this study, 219 isolates of these species were characterized by molecular identity analyses using BLAST of the ITS and EF1α genomic regions and by pathogenicity testing in outdoor, large-container trials. Molecular identity analyses confirmed the identity of isolates at 99 to 100% similarity to reference isolates in the database. In pathogenicity experiments, F. solani produced the most severe symptoms, followed by F. culmorum-like fungi, F. oxysporum, P. cucumerina, and S. terrestris. Some treatments of mixed species inoculum produced symptoms above what was expected from individual species. In particular, the mixture of F. culmorum-like fungi, F. oxysporum, and P. cucumerina and the mixture of F. culmorum-like fungi, F. solani, and S. terrestris had equally severe symptom ratings than that of F. solani by itself. Results indicate that this soilborne disease is primarily caused by Fusarium solani, but interactions among the complex of F. solani, F. culmorum-like fungi, F. oxysporum, and P. cucumerina, can exacerbate disease severity.


Author(s):  
Mohamed F. R Khan ◽  
Md. Ziaur Rahman Bhuyian ◽  
Dilip Lashman ◽  
Presley Mosher ◽  
Timothy Cymbaluk ◽  
...  

In this contribution, we report on a survey of leaf blight on sugar beet in the Sidney factory district of Montana caused by the soilborne fungus Sclerotinia sclerotiorum. In addition to morphological and pathological characterization of the pathogen on sugar beet, we confirmed the molecular identity of the fungus by DNA sequence analysis of the ribosomal Internal Transcribed Spacer region. Our findings are significant because the sugar beet production in south-central and eastern Montana contributes more than $100 million in economic activities. The incidence of this emerging disease will require changes in conventional disease management.


2021 ◽  
Author(s):  
Yann Roussel ◽  
Csaba Verasztó ◽  
Dimitri Rodarie ◽  
Tanguy Damart ◽  
Michael W Reimann ◽  
...  

Knowledge of the cell-type-specific composition of the brain is useful in order to understand the role of each cell type as part of the network. Here, we estimated the composition of the whole cortex in terms of well characterised morphological and electrophysiological inhibitory neuron types (me-types). We derived probabilistic me-type densities from an existing atlas of molecularly defined cell-type densities in the mouse cortex. We used a well-established me-type classification from rat somatosensory cortex to populate the cortex. These me-types were well characterized morphologically and electrophysiologically but they lacked molecular marker identity labels. To extrapolate this missing information, we employed an additional dataset from the Allen Institute for Brain Science containing molecular identity as well as morphological and electrophysiological data for mouse cortical neurons. We first built a latent space based on a number of comparable morphological and electrical features common to both data sources. We then identified 13 morpho-electrical clusters that merged neurons from both datasets while being molecularly homogeneous. The resulting clusters best mirror the molecular identity classification solely using available morpho-electrical features. Finally, we stochastically assigned a molecular identity to a me-type neuron based on the latent space cluster it was assigned to. The resulting mapping was used to derive inhibitory me-types densities in the cortex.


2021 ◽  
Vol 12 ◽  
Author(s):  
Yanfang Feng ◽  
Arend L. de Vos ◽  
Shakir Khan ◽  
Mary St. John ◽  
Tayyaba Hasan

Objectives: Carbapenemase-producing organisms (CPOs) are associated with high mortality rates. The recent development of β-lactamase inhibitors (BLIs) has made it possible to control CPO infections safely and effectively with β-lactams (BLs). This study aims to explicate the quantitative relationship between BLI’s β-lactamase inhibition and CPO’s BL susceptibility restoration, thereby providing the infectious disease society practical scientific grounds for regulating the use of BL/BLI in CPO infection treatment.Methods: A diverse collection of human CPO infection isolates was challenged by three structurally representative BLIs available in the clinic. The resultant β-lactamase inhibition, BL susceptibility restoration, and their correlation were followed quantitatively for each isolate by coupling FIBA (fluorescence identification of β-lactamase activity) and BL antibiotic susceptibility testing.Results: The β-lactamase inhibition and BL susceptibility restoration are positively correlated among CPOs under the treatment of BLIs. Both of them are dependent on the target CPO’s carbapenemase molecular identity. Of note, without sufficient β-lactamase inhibition, CPO’s BL susceptibility restoration is universally low across all tested carbapenemase molecular groups. However, a high degree of β-lactamase inhibition would not necessarily lead to a substantial BL susceptibility restoration in CPO probably due to the existence of non-β-lactamase BL resistance mechanisms.Conclusion: BL/BLI choice and dosing should be guided by quantitative tools that can evaluate the inhibition across the entire β-lactamase background of the CPO upon the BLI administion. Furthermore, rapid molecular diagnostics for BL/BLI resistances, especially those sensitive to β-lactamase independent BL resistance mechanisms, should be exploited to prevent ineffective BL/BLI treatment.


Parasitologia ◽  
2021 ◽  
Vol 1 (4) ◽  
pp. 247-256
Author(s):  
Anna Regina Angela Marquez ◽  
Kieran Eamens ◽  
Mark Westman ◽  
Jan Šlapeta

Rhipicephalus sanguineus s.l. and Ctenocephalides felis are considered the most prevalent ectoparasites of dogs in the Philippines. Vector-borne pathogens (VBPs) in these ectoparasites pose health risks to humans and animals. This study aimed to confirm the morphological and molecular identity of tick and flea species parasitising dogs in Metro Manila (Philippines) and molecularly investigate the possible presence of Bartonella spp., Rickettsia spp., Ehrlichia canis, and Anaplasma platys DNA. Ticks (n = 58) and fleas (n = 52) on dogs from three veterinary clinics in Metro Manila were collected and identified morphologically and molecularly via amplification and sequencing of cytochrome c oxidase I (cox1). Aliquots of ectoparasite DNA underwent real-time polymerase chain reaction (qPCR) screening for VBPs. All ticks were R. linnaei (formerly R. sanguineus s.l. “tropical lineage”), while all fleas were C. felis from clade 6 of the tropical II cluster/“Cairns” clade known from Australia. DNA of B. clarridgeiae was detected in 10% of fleas. DNA of R. felis was detected in 10% of fleas and in 3.8% of ticks. DNA of E. canis and A. platys was not detected. This study confirmed the presence of ticks and fleas as frequent ectoparasites on dogs and VBP presence emphasises the importance of preventative actions for animal health and welfare.


Author(s):  
Bhavani S. Sahu ◽  
Megin E. Nguyen ◽  
Pedro Rodriguez ◽  
Jean Pierre Pallais ◽  
Vinayak Ghosh ◽  
...  

AbstractThe TLQP-21 neuropeptide has been implicated in functions as diverse as lipolysis, neurodegeneration and metabolism, thus suggesting an important role in several human diseases. Three binding targets have been proposed for TLQP-21: C3aR1, gC1qR and HSPA8. The aim of this review is to critically evaluate the molecular identity of the TLQP-21 receptor and the proposed multi-receptor mechanism of action. Several studies confirm a critical role for C3aR1 in TLQP-21 biological activity and a largely conserved mode of binding, receptor activation and signaling with C3a, its first-identified endogenous ligand. Conversely, data supporting a role of gC1qR and HSPA8 in TLQP-21 activity remain limited, with no signal transduction pathways being described. Overall, C3aR1 is the only receptor for which a necessary and sufficient role in TLQP-21 activity has been confirmed thus far. This conclusion calls into question the validity of a multi-receptor mechanism of action for TLQP-21 and should inform future studies.


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