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1.
Curr Protoc ; 4(3): e978, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38511467

RESUMO

16S rRNA targeted amplicon sequencing is an established standard for elucidating microbial community composition. While high-throughput short-read sequencing can elicit only a portion of the 16S rRNA gene due to their limited read length, third generation sequencing can read the 16S rRNA gene in its entirety and thus provide more precise taxonomic classification. Here, we present a protocol for generating full-length 16S rRNA sequences with Oxford Nanopore Technologies (ONT) and a microbial community profile with Emu. We select Emu for analyzing ONT sequences as it leverages information from the entire community to overcome errors due to incomplete reference databases and hardware limitations to ultimately obtain species-level resolution. This pipeline provides a low-cost solution for characterizing microbiome composition by exploiting real-time, long-read ONT sequencing and tailored software for accurate characterization of microbial communities. © 2024 Wiley Periodicals LLC. Basic Protocol: Microbial community profiling with Emu Support Protocol 1: Full-length 16S rRNA microbial sequences with Oxford Nanopore Technologies sequencing platform Support Protocol 2: Building a custom reference database for Emu.


Assuntos
Dromaiidae , Microbiota , Animais , RNA Ribossômico 16S/genética , Dromaiidae/genética , Bactérias/genética , Análise de Sequência de DNA/métodos , Microbiota/genética
2.
J Comp Neurol ; 532(2): e25587, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38335048

RESUMO

We examined the presence/absence and parcellation of cholinergic neurons in the hypothalami of five birds: a Congo grey parrot (Psittacus erithacus), a Timneh grey parrot (P. timneh), a pied crow (Corvus albus), a common ostrich (Struthio camelus), and an emu (Dromaius novaehollandiae). Using immunohistochemistry to an antibody raised against the enzyme choline acetyltransferase, hypothalamic cholinergic neurons were observed in six distinct clusters in the medial, lateral, and ventral hypothalamus in the parrots and crow, similar to prior observations made in the pigeon. The expression of cholinergic nuclei was most prominent in the Congo grey parrot, both in the medial and lateral hypothalamus. In contrast, no evidence of cholinergic neurons in the hypothalami of either the ostrich or emu was found. It is known that the expression of sleep states in the ostrich is unusual and resembles that observed in the monotremes that also lack hypothalamic cholinergic neurons. It has been proposed that the cholinergic system acts globally to produce and maintain brain states, such as those of arousal and rapid-eye-movement sleep. The hiatus in the cholinergic system of the ostrich, due to the lack of hypothalamic cholinergic neurons, may explain, in part, the unusual expression of sleep states in this species. These comparative anatomical and sleep studies provide supportive evidence for global cholinergic actions and may provide an important framework for our understanding of one broad function of the cholinergic system and possible dysfunctions associated with global cholinergic neural activity.


Assuntos
Dromaiidae , Struthioniformes , Animais , Dromaiidae/metabolismo , Struthioniformes/metabolismo , Encéfalo/metabolismo , Hipotálamo/metabolismo , Neurônios Colinérgicos/metabolismo , Sono/fisiologia , Colinérgicos , Colina O-Acetiltransferase/metabolismo
3.
PLoS One ; 19(2): e0298785, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38354182

RESUMO

The vibration and radiation noise characteristics of the gear transmission system are different under different traction conditions, and the gear modification optimization scheme based on a single working condition is not suitable for the operating environment under all working conditions. To modify the traction gear of a high-speed EMU, an optimized design scheme for noise reduction under multiple working conditions is proposed. A modification plan of the tooth direction in conjunction with the tooth shape was devised using a parametric model of the EMU's traction gear transmission system. The radiation noise of the gear transmission system after modification was solved using the acoustic boundary element method under different working conditions. A gear noise prediction model based on the random forest was proposed, and a gear modification parameter combination was constructed to minimize radiation noise. Then, the optimal design scheme of multi-condition modification combination parameters is obtained with the weight of the running time and acoustic contribution under different working conditions. The grey correlation degree evaluation model is established to verify that the multi-condition modification optimization design method can make the traction gear of EMU obtain satisfactory transmission performance and noise reduction effect under different working conditions.


Assuntos
Dromaiidae , Tração , Animais , Acústica , Vibração , Condições de Trabalho
4.
Artigo em Inglês | MEDLINE | ID: mdl-38220129

RESUMO

The baroreflex involves cardiovascular homeostatic mechanisms that buffer the system against acute deviations in arterial blood pressure. It is comprised of the cardiac limb which involves adjustments in heart rate and the peripheral limb which involves adjustments in vascular resistance. This negative feedback loop mechanism has been investigated in numerous species of adult vertebrates, however our understanding of the maturation and functional importance of the reflex in developing animals remains poorly understood. In egglaying species, our knowledge of this mechanism is limited to the domestic chicken embryo and the embryonic alligator. While each of these species possess a cardiac baroreflex prior to hatching, they differ in the timing when it becomes functional, with the embryonic chicken possessing the reflex at 90% of incubation, while the alligator possesses the reflex at 70% of incubation. In an effort to determine if bird species might share similar patterns of active baroreflex function, we studied embryonic emus (Dromiceius novaehollandiae). However, we hypothesized that emus would possess a pattern of baroreflex function similar to that of the American alligator given the emu embryo possesses functional vagal tone at 70% of incubation, possibly indicating a more mature collection of cardiovascular control mechanism than those found in embryonic chickens. Our findings illustrate that emu embryos possess a hypotensive baroreflex at 90% of incubation. Therefore, our data fail to support our original hypothesis. While only two species of birds have been studied in this context, it could indicate that baroreflex function is not essential for cardiovascular homeostasis in birds for the majority of in ovo development.


Assuntos
Sistema Cardiovascular , Dromaiidae , Embrião de Galinha , Animais , Barorreflexo/fisiologia , Galinhas , Pressão Arterial , Frequência Cardíaca/fisiologia , Pressão Sanguínea/fisiologia
5.
Anat Rec (Hoboken) ; 306(7): 1842-1863, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37314297

RESUMO

The late Miocene Dromornis stirtoni is the largest of the giant flightless dromornithid birds. Here, we studied 22 long bones (femora, tibiotarsi, tarsometatarsi) of D. stirtoni to assess its osteohistology to deduce various aspects of its life history. Our results show that D. stirtoni took several years (likely, more than a decade), to reach adult body size, after which its growth rate slowed down, and skeletal maturity occurred. This growth strategy differs from that of its Pleistocene relative, Genyornis newtoni, which experienced faster rates of growth to reach adult body size. We propose that these mihirung birds, separated by millions of years, each responded to the prevailing environmental conditions of the time, by selecting for different growth strategies, with D. stirtoni having an extreme K-selected life history strategy. The presence of medullary bone permitted the identification of female D. stirtoni specimens, and its presence in some bones lacking an OCL layer showed sexual maturity preceded its formation. We postulate that while G. newtoni had a somewhat greater reproductive potential than D. stirtoni, it remained far less than that observed in the extant emu (Dromaius novaehollandiae). Genyornis newtoni survived into the late Pleistocene alongside extant emus and overlapped the arrival of the first humans in Australia, but the former species shortly thereafter became extinct while emus remain prolific.


Assuntos
Osso e Ossos , Dromaiidae , Animais , Feminino , Austrália
6.
Epilepsy Behav ; 144: 109252, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37207403

RESUMO

INTRODUCTION AND BACKGROUND: The impact of the timing of the last seizure (TTLS) prior to admission to the epilepsy monitoring unit (EMU) on epilepsy classification is unclear for which we conducted this study. METHODS: We reviewed patients with epilepsy admitted to EMU between January 2021 and April 2022 and identified TTLS before EMU admission. We considered EMU yield as whether; it confirmed epilepsy classification, added new knowledge to the classification, or failed to classify epilepsy. RESULTS: We studied 156 patients. There were 72 (46%) men, with a mean age of 30. TTLS was divided according to a one- or three-month cutoff. We confirmed the pre-EMU epilepsy classification in 52 (33%) patients, learned new findings on epilepsy classification in 80 (51%) patients, and failed to classify epilepsy in 24 (15%) patients. Patients with "confirmed epilepsy classifications" reported seizures sooner to EMU admission than other groups (0.7 vs. 2.3 months, p-value = 0.02, 95% CI; -1.8, -1.3). Also, the odds of confirming epilepsy classification were more than two times in patients with TTLS within a month compared to those with TTLS of more than a month (OR = 2.4, p-value = 0.04, 95% CI; 1.1, 5.9). The odds were also higher when the 3-month TTLS cutoff was considered (OR = 6.2, p-value = 0.002, 95% CI; 1.6, 40.2). Confirming epilepsy classification was also associated with earlier seizures recorded at one- or three-month cutoff (OR = 2.1 and OR = 2.3, respectively, p-value = 0.05). We did not observe similar findings when we modified the classification or failed to reach a classification. CONCLUSIONS: The timing of the last seizure before EMU admission appeared to influence the yield of EMU and enhanced the confirmation of epilepsy classifications. Such findings can improve the utilization of EMU in the presurgical evaluation of patients with epilepsy.


Assuntos
Dromaiidae , Epilepsia , Masculino , Animais , Humanos , Adulto , Feminino , Eletroencefalografia , Estudos Retrospectivos , Convulsões/diagnóstico , Epilepsia/complicações , Epilepsia/diagnóstico , Epilepsia/cirurgia , Monitorização Fisiológica
7.
BMJ Open ; 13(4): e070236, 2023 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-37012020

RESUMO

INTRODUCTION: People who inject drugs (PWID) are at risk of invasive infections such as bloodstream infections, endocarditis, osteomyelitis and septic arthritis. Such infections require prolonged antibiotic therapy, but there is limited evidence about the optimal care model to deliver to this population. The Epidemiology and Management of invasive infections among people who Use drugs (EMU) study aims to (1) describe the current burden, clinical spectrum, management and outcomes of invasive infections in PWID; (2) determine the impact of currently available models of care on completion of planned antimicrobials for PWID admitted to hospital with invasive infections and (3) determine postdischarge outcomes of PWID admitted with invasive infections at 30 and 90 days. METHODS AND ANALYSIS: EMU is a prospective multicentre cohort study of Australian public hospitals who provide care to PWIDs with invasive infections. All patients who have injected drugs in the previous six months and are admitted to a participating site for management of an invasive infection are eligible. EMU has two components: (1) EMU-Audit will collect information from medical records, including demographics, clinical presentation, management and outcomes; (2) EMU-Cohort will augment this with interviews at baseline, 30 and 90 days post-discharge, and data linkage examining readmission rates and mortality. The primary exposure is antimicrobial treatment modality, categorised as inpatient intravenous antimicrobials, outpatient antimicrobial therapy, early oral antibiotics or lipoglycopeptide. The primary outcome is confirmed completion of planned antimicrobials. We aim to recruit 146 participants over a 2-year period. ETHICS AND DISSEMINATION: EMU has been approved by the Alfred Hospital Human Research Ethics Committee (Project number 78815.) EMU-Audit will collect non-identifiable data with a waiver of consent. EMU-Cohort will collect identifiable data with informed consent. Findings will be presented at scientific conferences and disseminated by peer-review publications. TRIAL REGISTRATION NUMBER: ACTRN12622001173785; Pre-results.


Assuntos
Dromaiidae , Abuso de Substâncias por Via Intravenosa , Humanos , Animais , Assistência ao Convalescente , Abuso de Substâncias por Via Intravenosa/complicações , Abuso de Substâncias por Via Intravenosa/epidemiologia , Estudos Prospectivos , Estudos de Coortes , Alta do Paciente , Austrália/epidemiologia , Antibacterianos/uso terapêutico , Estudos Multicêntricos como Assunto
8.
Sci Rep ; 13(1): 4488, 2023 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-36934111

RESUMO

Emus (Dromaius novaehollandiae), a large flightless omnivorous ratite, are farmed for their fat and meat. Emu fat can be rendered into oil for therapeutic and cosmetic use. They are capable of gaining a significant portion of its daily energy requirement from the digestion of plant fibre. Despite of its large body size and low metabolic rate, emus have a relatively simple gastroinstetinal (GI) tract with a short mean digesta retention time. However, little is known about the GI microbial diversity of emus. The objective of this study was to characterize the intraluminal intestinal bacterial community in the different segments of small intestine (duodenum, jejunum, and ileum) using pyrotag sequencing and compare that with the ceca. Gut content samples were collected from each of four adult emus (2 males, 2 females; 5-6 years old) that were free ranged but supplemented with a barley-alfalfa-canola based diet. We amplified the V3-V5 region of 16S rRNA gene to identify the bacterial community using Roche 454 Junior system. After quality trimming, a total of 165,585 sequence reads were obtained from different segments of the small intestine (SI). A total of 701 operational taxonomic units (OTUs) were identified in the different segments of small intestine. Firmicutes (14-99%) and Proteobacteria (0.5-76%) were the most predominant bacterial phyla in the small intestine. Based on species richness estimation (Chao1 index), the average number of estimated OTUs in the small intestinal compartments were 148 in Duodenum, 167 in Jejunum, and 85 in Ileum, respectively. Low number of core OTUs identified in each compartment of small intestine across individual birds (Duodenum: 13 OTUs, Jejunum: 2 OTUs, Ileum: 14 OTUs) indicated unique bacterial community in each bird. Moreover, only 2 OTUs (Escherichia and Sinobacteraceae) were identified as core bacteria along the whole small intestine. PICRUSt analysis has indicated that the detoxification of plant material and environmental chemicals seem to be performed by SI microbiota, especially those in the jejunum. The emu cecal microbiome has more genes than SI segments involving in protective or immune response to enteric pathogens. Microbial digestion and fermentation is mostly in the jejunum and ceca. This is the first study to characterize the microbiota of different compartments of the emu intestines via gut samples and not fecal samples. Results from this study allow us to further investigate the influence of the seasonal and physiological changes of intestinal microbiota on the nutrition of emus and indirectly influence the fatty acid composition of emu fat.


Assuntos
Dromaiidae , Jejuno , Animais , Masculino , Feminino , Jejuno/microbiologia , RNA Ribossômico 16S/genética , Duodeno , Íleo , Bactérias/genética
9.
Spectrochim Acta A Mol Biomol Spectrosc ; 294: 122506, 2023 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-36868023

RESUMO

Consumers demand safe and nutritious foods at accessible prices; where issues associated with adulteration, fraud, and provenance have become important aspects to be considered by the modern food industry. There are many analytical techniques and methods available to determine food composition and quality, including food security. Among them, vibrational spectroscopy techniques are at the first line of defence (near and mid infrared spectroscopy, and Raman spectroscopy). In this study, a portable near infrared (NIR) instrument was evaluated to identify different levels of adulteration between binary mixtures of exotic and traditional meat species. Fresh meat cuts of lamb (Ovis aries), emu (Dromaius novaehollandiae), camel (Camelus dromedarius) and beef (Bos taurus) sourced from a commercial abattoir were used to make different binary mixtures (95 % %w/w, 90 % %w/w, 50 % %w/w, 10 % %w/w and 5 % %w/w) and analysed using a portable NIR instrument. The NIR spectra of the meat mixtures was analysed using principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA). Two isosbestic points corresponding to absorbances at 1028 nm and 1224 nm were found to be consistent across all the binary mixtures analysed. The coefficient of determination in cross validation (R2) obtained for the determination of the per cent of species in a binary mixture was above 90 % with a standard error in cross validation (SECV) ranging between 12.6 and 15 %w/w. Overall, the results of this study indicate that NIR spectroscopy can determine the level or ratio of adulteration in the binary mixtures of minced meat.


Assuntos
Dromaiidae , Carneiro Doméstico , Ovinos , Bovinos , Animais , Camelus , Quimiometria , Contaminação de Alimentos/análise , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Análise dos Mínimos Quadrados
10.
Br Poult Sci ; 64(3): 435-440, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36607349

RESUMO

1. Sex chromosomes of emus are largely homomorphic. Therefore, the standard methodology for molecular sexing based on screening intron length variations in sex-linked genes is not applicable. However, emu sexing requires costly and time-consuming PCR-RFLP or multiplex PCR methods.2. This experiment used a directed PCR amplification and capillary electrophoresis sexing protocol. Two distinct peaks were observed in females (ZW), while only one peak was observed in males (ZZ).3. This sexing technique proved to be rapid, non-invasive, and highly sensitive and may be useful for verifying the sex ratio and breeding management of emus.


Assuntos
Dromaiidae , Feminino , Masculino , Animais , Dromaiidae/genética , Galinhas/genética , Polimorfismo de Fragmento de Restrição , Íntrons
11.
Anim Sci J ; 93(1): e13786, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36464246

RESUMO

Emus (Dromaius novaehollandiae) are expected to become a novel poultry species for producing eggs, meat, and oil. In our previous studies, Japanese emu populations were predicted to have reduced genetic diversity through inbreeding. For a sustainable emu industry in Japan, it is necessary to understand the current genetic structure and relationships in dispersed farms. In this study, we investigated the genetic structure and relationships of six Japanese emu farms based on mitochondrial DNA and microsatellite polymorphisms. We analyzed the DNA sequences of the mitochondrial D-loop region in 157 individuals and detected four haplotypes with four nucleotide substitution sites (Hap-a, Hap-b, Hap-c, and Hap-d). Analysis of molecular variance revealed that 43.6% of total variance was "among population," and the FST value was 0.436 with significant genetic differentiation (P < 0.001). In microsatellite analysis, the expected (HE ) and observed (HO ) heterozygosities were 0.53-0.64 and 0.44-0.59, respectively. Phylogenetic trees and STRUCTURE analysis revealed that the six Japanese farmed emu populations could be divided into four genetically differentiated groups. Therefore, we identified genetic resources that may be useful in extending the genetic diversity of Japanese farms and are predicted to contribute to the conservation and reconstruction of populations.


Assuntos
Dromaiidae , Animais , Dromaiidae/genética , Fazendas , Japão , Filogenia , Óvulo , DNA Mitocondrial/genética , Repetições de Microssatélites/genética
12.
Poult Sci ; 101(10): 102050, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35952603

RESUMO

Characterization of carcass traits and fat quality is important to effectively produce and genetically improve emus. We investigated carcass traits in 309 emus. The meat production of female emus showed a significantly higher value than that of males (P < 0.01). The fat weight of male (9.232 ± 3.156 kg) was larger than that of the female (7.772 ± 2.697 kg). The fat yield (fat weight per kg of body weight) was strongly correlated to body weight (r = 0.79 and r = 0.75 in male and female, respectively). The fat melting points of females and males were 19.19 ± 3.39°C and 19.39 ± 3.39°C, respectively, without significant difference. Since the fat melting point did not correlate to body and fat weights, we predicted that it was an independent trait from body growth and was highly influenced by genetic elements. Percentages of palmitic, stearic, oleic, linoleic, and α-linolenic acids were 22.27 ± 3.50%, 9.37 ± 1.90%, 54.11 ± 5.17%, 13.54 ± 7.80% and 0.71 ± 0.59%, respectively. Among them, linoleic acid contents showed a wide individual difference (range 0.3-19.9%). The oleic/stearic acid ratio showed a negative correlation to the fat melting point. These results suggest that the fat melting point is a good indicator of C18:1/C18:0 ratio in emu fat.


Assuntos
Dromaiidae , Animais , Composição Corporal/genética , Peso Corporal/genética , Galinhas , Ácidos Graxos , Feminino , Japão , Ácidos Linoleicos , Ácidos Linolênicos , Masculino , Carne/análise , Ácidos Esteáricos
13.
Biol Pharm Bull ; 45(8): 1022-1026, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35908885

RESUMO

The emu is the second largest ratite; thus, their sera and egg yolks, obtained after immunization, could provide therapeutic and diagnostically important immunoglobulins with improved production efficiency. Reliable purification tools are required to establish a pipeline for supplying practical emu-derived antibodies, the majority of which belongs to the immunoglobulin Y (IgY) class. Therefore, we generated a monoclonal secondary antibody specific to emu IgY. Initially, we immunized an emu with bovine serum albumin multiply haptenized with 2,4-dinitrophenyl (DNP) groups. Polyclonal emu anti-DNP antibodies were partially purified using conventional precipitation method and used as antigen for immunizing a BALB/c mouse. Splenocytes were fused with myeloma cells and a hybridoma clone secreting a desirable secondary antibody (mAb#2-16) was established. The secondary antibody bound specifically to emu-derived IgY, distinguishing IgYs from chicken, duck, ostrich, quail, and turkey, as well as human IgGs. Affinity columns immobilizing the mAb#2-16 antibodies enabled purification of emu IgY fractions from sera and egg yolks via simple protocols, with which we succeeded in producing IgYs specific to the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) spike protein with a practical binding ability. We expect that the presented purification method, and the secondary antibody produced in this study, will facilitate the utilization of emus as a novel source of therapeutic and diagnostic antibodies.


Assuntos
COVID-19 , Dromaiidae , Animais , Anticorpos Monoclonais , Teste para COVID-19 , Galinhas/metabolismo , Dromaiidae/metabolismo , Humanos , Imunoglobulinas , Camundongos , SARS-CoV-2
14.
Sci Rep ; 12(1): 9419, 2022 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-35676317

RESUMO

Emu (Dromaius novaehollandiae) farming has been gaining wide interest for fat production. Oil rendered from this large flightless bird's fat is valued for its anti-inflammatory and antioxidant properties for uses in therapeutics and cosmetics. We analyzed the seasonal and sex-dependent differentially expressed (DE) genes involved in fat metabolism in emus. Samples were taken from back and abdominal fat tissues of a single set of four male and four female emus in April, June, and November for RNA-sequencing. We found 100 DE genes (47 seasonally in males; 34 seasonally in females; 19 between sexes). Seasonally DE genes with significant difference between the sexes in gene ontology terms suggested integrin beta chain-2 (ITGB2) influences fat changes, in concordance with earlier studies. Six seasonally DE genes functioned in more than two enriched pathways (two female: angiopoietin-like 4 (ANGPTL4) and lipoprotein lipase (LPL); four male: lumican (LUM), osteoglycin (OGN), aldolase B (ALDOB), and solute carrier family 37 member 2 (SLC37A2)). Two sexually DE genes, follicle stimulating hormone receptor (FSHR) and perilipin 2 (PLIN2), had functional investigations supporting their influence on fat gain and loss. The results suggested these nine genes influence fat metabolism and deposition in emus.


Assuntos
Dromaiidae , Tecido Adiposo/metabolismo , Animais , Sequência de Bases , Dromaiidae/genética , Feminino , Expressão Gênica , Masculino , Estações do Ano
15.
Nat Methods ; 19(7): 845-853, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35773532

RESUMO

16S ribosomal RNA-based analysis is the established standard for elucidating the composition of microbial communities. While short-read 16S rRNA analyses are largely confined to genus-level resolution at best, given that only a portion of the gene is sequenced, full-length 16S rRNA gene amplicon sequences have the potential to provide species-level accuracy. However, existing taxonomic identification algorithms are not optimized for the increased read length and error rate often observed in long-read data. Here we present Emu, an approach that uses an expectation-maximization algorithm to generate taxonomic abundance profiles from full-length 16S rRNA reads. Results produced from simulated datasets and mock communities show that Emu is capable of accurate microbial community profiling while obtaining fewer false positives and false negatives than alternative methods. Additionally, we illustrate a real-world application of Emu by comparing clinical sample composition estimates generated by an established whole-genome shotgun sequencing workflow with those returned by full-length 16S rRNA gene sequences processed with Emu.


Assuntos
Dromaiidae , Microbiota , Sequenciamento por Nanoporos , Animais , Bactérias/genética , Dromaiidae/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Microbiota/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodos
16.
J Anat ; 241(2): 518-526, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35412666

RESUMO

Ostriches and emus are among the largest extant birds and are frequently used as modern analogs for the growth dynamics of non-avian theropod dinosaurs. These ratites quickly reach adult size in under 1 year, and as such do not typically exhibit annually deposited growth marks. Growth marks, commonly classified as annuli or lines of arrested growth (LAGs), represent reduced or halted osteogenesis, respectively, and their presence demonstrates varying degrees of developmental plasticity. Growth marks have not yet been reported from ostriches and emus, prompting authors to suggest that they have lost the plasticity required to deposit them. Here we observe the hind limb bone histology of three captive juvenile emus and one captive adult ostrich. Two of the three juvenile emus exhibit typical bone histology but the third emu, a 4.5-month-old juvenile, exhibits a regional arc of avascular tissue, which we interpret as a growth mark. As this mark is not present in the other two emus from the same cohort and it co-occurs with a contralateral broken fibula, we suggest variable biomechanical load as a potential cause. The ostrich exhibits a complete ring of avascular, hypermineralized bone with sparse, flattened osteocyte lacunae. We identify this as an annulus and interpret it as slowing of growth. In the absence of other growth marks and lacking the animal's life history, the timing and cause of this ostrich's reduced growth are unclear. Even so, these findings demonstrate that both taxa retain the ancestral developmental plasticity required to temporarily slow growth. We also discuss the potential challenges of identifying growth marks using incomplete population data sets and partial cortical sampling.


Assuntos
Dinossauros , Dromaiidae , Paleógnatas , Struthioniformes , Animais , Osso e Ossos , Humanos
17.
Sci Rep ; 12(1): 6325, 2022 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-35428830

RESUMO

Emus are farmed for fat production. Oil rendered from their back and abdominal fat pads has good anti-oxidant and anti-inflammatory properties and has ingredients that promote cell growth. Our objective is to examine the mRNA expression of 7 emu adipokine genes (eFABP4, eSCD1, eAdipoQ, eAdipoR1, eAdipoR2, eLEP and eLepR) to identify gene markers that may help improve emu fat production. Back and abdominal fat tissues from 11 adult emus were biopsied at four time points (April, June, August and November). Total RNA was isolated and cDNA was synthesized. Gene specific primers were designed for partial cloning fragments to amplify the open reading frame of the 7 genes. eLEP was not expressed in emu fat tissue. Nucleotides and amino acids sequences of the 6 expressed gene were compared with homologs from other species and phylogenetic relationships established. Seasonal mRNA expression of each gene was assessed by quantitative RT-PCR and differential expression analysed by the 2-ΔΔCT method. The 6 expressed genes showed seasonal variation in expression and showed association of expression level with back fat adiposity. More whole-genome scanning studies are needed to develop novel molecular markers that can be applied to improve fat production in emus.


Assuntos
Dromaiidae , Adipocinas/genética , Animais , Filogenia , RNA Mensageiro/genética , Estações do Ano
18.
Seizure ; 96: 13-17, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35042003

RESUMO

PURPOSE: We assessed whether automated detection software, combined with live observation, enabled reliable seizure detection using three commercial software packages: Persyst, Encevis and BESA. METHODS: Two hundred and eighty-six prolonged EEG records of individuals aged 16-86 years, collected between August 2019 and January 2020, were retrospectively processed using all three packages. The reference standard included all seizures mentioned in the clinical report supplemented with true detections made by the software and not previously detected by clinical physiologists. Sensitivity was measured for offline review by clinical physiologists and software seizure detection, both in combination with live monitoring in an EMU setting, for all three software packages at record and seizure level. RESULTS: The database contained 249 seizures in 64 records. The sensitivity of seizure detection was 98% for Encevis and Persyst, and 95% for BESA, when a positive results was defined as detection at least one of the seizures occurring within an individual record. When positivity was defined as recognition of all seizures, sensitivity was 93% for Persyst, 88% for Encevis and 84% for BESA. Clinical physiologists' review had a sensitivity of 100% at record level and 98% at seizure level. The median false positive rate per record was 1.7 for Persyst, 2.4 for BESA and 5.5 for Encevis per 24 h. CONCLUSION: Automated seizure detection software does not perform as well as technicians do. However, it can be used in an EMU setting when the user is aware of its weaknesses. This assessment gives future users helpful insight into these strengths and weaknesses. The Persyst software performs best.


Assuntos
Dromaiidae , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Eletroencefalografia/métodos , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Convulsões/diagnóstico , Software , Adulto Jovem
20.
Blood Adv ; 5(14): 2817-2828, 2021 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-34269799

RESUMO

Chronic lymphocytic leukemia (CLL) is caused by the progressive accumulation of mature CD5+ B cells in secondary lymphoid organs. In vitro data suggest that CD4+ T lymphocytes also sustain survival and proliferation of CLL clones through CD40L/CD40 interactions. In vivo data in animal models are conflicting. To clarify this clinically relevant biological issue, we generated genetically modified Eµ-TCL1 mice lacking CD4+ T cells (TCL1+/+AB0), CD40 (TCL1+/+CD40-/-), or CD8+ T cells (TCL1+/+TAP-/-), and we monitored the appearance and progression of a disease that mimics aggressive human CLL by flow cytometry and immunohistochemical analyses. Findings were confirmed by adoptive transfer of leukemic cells into mice lacking CD4+ T cells or CD40L or mice treated with antibodies depleting CD4 T cells or blocking CD40L/CD40 interactions. CLL clones did not proliferate in mice lacking or depleted of CD4+ T cells, thus confirming that CD4+ T cells are essential for CLL development. By contrast, CD8+ T cells exerted an antitumor activity, as indicated by the accelerated disease progression in TCL1+/+TAP-/- mice. Antigen specificity of CD4+ T cells was marginal for CLL development, because CLL clones efficiently proliferated in transgenic mice whose CD4 T cells had a T-cell receptor with CLL-unrelated specificities. Leukemic clones also proliferated when transferred into wild-type mice treated with monoclonal antibodies blocking CD40 or into CD40L-/- mice, and TCL1+/+CD40-/- mice developed frank CLL. Our data demonstrate that CD8+ T cells restrain CLL progression, whereas CD4+ T cells support the growth of leukemic clones in TCL1 mice through CD40-independent and apparently noncognate mechanisms.


Assuntos
Dromaiidae , Leucemia Linfocítica Crônica de Células B , Animais , Linfócitos T CD4-Positivos , Ligante de CD40/genética , Leucemia Linfocítica Crônica de Células B/genética , Camundongos , Camundongos Transgênicos , Proteínas Proto-Oncogênicas
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