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In recent years, there has been a surge in the publication of clinical trial reports, making it challenging to conduct systematic reviews. Automatically extracting Population, Intervention, Comparator, and Outcome (PICO) from clinical trial studies can alleviate the traditionally time-consuming process of manually scrutinizing systematic reviews. Existing approaches of PICO frame extraction involves supervised approach that relies on the existence of manually annotated data points in the form of BIO label tagging. Recent approaches, such as In-Context Learning (ICL), which has been shown to be effective for a number of downstream NLP tasks, require the use of labeled examples. In this work, we adopt ICL strategy by employing the pretrained knowledge of Large Language Models (LLMs), gathered during the pretraining phase of an LLM, to automatically extract the PICO-related terminologies from clinical trial documents in unsupervised set up to bypass the availability of large number of annotated data instances. Additionally, to showcase the highest effectiveness of LLM in oracle scenario where large number of annotated samples are available, we adopt the instruction tuning strategy by employing Low Rank Adaptation (LORA) to conduct the training of gigantic model in low resource environment for the PICO frame extraction task. More specifically, both of the proposed frameworks utilize AlpaCare as base LLM which employs both few-shot in-context learning and instruction tuning techniques to extract PICO-related terms from the clinical trial reports. We applied these approaches to the widely used coarse-grained datasets such as EBM-NLP, EBM-COMET and fine-grained datasets such as EBM-NLPrev and EBM-NLPh. Our empirical results show that our proposed ICL-based framework produces comparable results on all the version of EBM-NLP datasets and the proposed instruction tuned version of our framework produces state-of-the-art results on all the different EBM-NLP datasets. Our project is available at https://github.com/shrimonmuke0202/AlpaPICO.git.
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Ensaios Clínicos como Assunto , Processamento de Linguagem Natural , Humanos , Ensaios Clínicos como Assunto/métodos , Mineração de Dados/métodos , Aprendizado de MáquinaRESUMO
PURPOSE: Large Language Models (LLMs) like Generative Pre-trained Transformer (GPT) from OpenAI and LLaMA (Large Language Model Meta AI) from Meta AI are increasingly recognized for their potential in the field of cheminformatics, particularly in understanding Simplified Molecular Input Line Entry System (SMILES), a standard method for representing chemical structures. These LLMs also have the ability to decode SMILES strings into vector representations. METHOD: We investigate the performance of GPT and LLaMA compared to pre-trained models on SMILES in embedding SMILES strings on downstream tasks, focusing on two key applications: molecular property prediction and drug-drug interaction prediction. RESULTS: We find that SMILES embeddings generated using LLaMA outperform those from GPT in both molecular property and DDI prediction tasks. Notably, LLaMA-based SMILES embeddings show results comparable to pre-trained models on SMILES in molecular prediction tasks and outperform the pre-trained models for the DDI prediction tasks. CONCLUSION: The performance of LLMs in generating SMILES embeddings shows great potential for further investigation of these models for molecular embedding. We hope our study bridges the gap between LLMs and molecular embedding, motivating additional research into the potential of LLMs in the molecular representation field. GitHub: https://github.com/sshaghayeghs/LLaMA-VS-GPT .
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Quimioinformática , Quimioinformática/métodos , Interações Medicamentosas , Estrutura MolecularRESUMO
BACKGROUND: Malnutrition is a prevalent issue in aged care facilities (RACFs), leading to adverse health outcomes. The ability to efficiently extract key clinical information from a large volume of data in electronic health records (EHR) can improve understanding about the extent of the problem and developing effective interventions. This research aimed to test the efficacy of zero-shot prompt engineering applied to generative artificial intelligence (AI) models on their own and in combination with retrieval augmented generation (RAG), for the automating tasks of summarizing both structured and unstructured data in EHR and extracting important malnutrition information. METHODOLOGY: We utilized Llama 2 13B model with zero-shot prompting. The dataset comprises unstructured and structured EHRs related to malnutrition management in 40 Australian RACFs. We employed zero-shot learning to the model alone first, then combined it with RAG to accomplish two tasks: generate structured summaries about the nutritional status of a client and extract key information about malnutrition risk factors. We utilized 25 notes in the first task and 1,399 in the second task. We evaluated the model's output of each task manually against a gold standard dataset. RESULT: The evaluation outcomes indicated that zero-shot learning applied to generative AI model is highly effective in summarizing and extracting information about nutritional status of RACFs' clients. The generated summaries provided concise and accurate representation of the original data with an overall accuracy of 93.25%. The addition of RAG improved the summarization process, leading to a 6% increase and achieving an accuracy of 99.25%. The model also proved its capability in extracting risk factors with an accuracy of 90%. However, adding RAG did not further improve accuracy in this task. Overall, the model has shown a robust performance when information was explicitly stated in the notes; however, it could encounter hallucination limitations, particularly when details were not explicitly provided. CONCLUSION: This study demonstrates the high performance and limitations of applying zero-shot learning to generative AI models to automatic generation of structured summarization of EHRs data and extracting key clinical information. The inclusion of the RAG approach improved the model performance and mitigated the hallucination problem.
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Inteligência Artificial , Registros Eletrônicos de Saúde , Humanos , Armazenamento e Recuperação da Informação/métodos , Desnutrição , Algoritmos , AustráliaRESUMO
A series of new hybrid compounds was prepared combining tetrahydropyran rings with different aromatic systems by means of a 1,2,3-triazole, using a building block strategy. The design of these structures was guided by Lead-Likeness and Molecular Analysis (LLAMA) software, adding modifications to our most potent scaffold (the tetrahydropyran ring) to generate promising "lead-like" candidates, which were subsequently compared against reported anticancer compounds. Our synthesized compounds demonstrated significant antiproliferative activity when compared with the standards cisplatin and 5-fluorouracil, across a panel of six different tumor cell lines. Moreover, compared with our group's previous hybrid compounds, these new structures exhibit similar activity while offering simpler synthesis and greater potential for structural diversification, a fact that was previously an issue. Further investigations on the most active compounds included assessments of reproductive cell survival, inhibition of cell migration, and effects on nuclear morphology, indicating potential diverse mechanisms of action for these compounds. Pharmacokinetic properties were also calculated for the whole series of compounds using the pkCSM online software.
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Hospital librarians receive invites to teach thinking and searching in an evidence-based way and critical appraisal of the literature to nurses. With these invitations, the hospital librarians play a central role in establishing an evidence-based culture in the hospital and contribute to the nursing staff feeling competent and confident in fulfilling evidence-based competencies. This author just prepared a 17-minute online talk as part of an international nursing webinar on "searching nursing literature in an evidence-based way." Using this experience, remembering other teaching and presentation experiences, and some "help" from AI tools, this experienced hospital librarian suggests decision points for colleagues to create a meaningful, practical information session for nurses and introduce to some AI tools along the way.
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Prática Clínica Baseada em Evidências , Bibliotecários , Humanos , Prática Clínica Baseada em Evidências/educaçãoRESUMO
BACKGROUND: Alpaca (Vicugna pacos), llama (Lama glama), vicugna (Vicugna vicugna) and guanaco (Lama guanicoe), are the camelid species distributed over the Andean high-altitude grasslands, the Altiplano, and the Patagonian arid steppes. Despite the wide interest on these animals, most of the loci under selection are still unknown. Using whole-genome sequencing (WGS) data we investigated the occurrence and the distribution of Runs Of Homozygosity (ROHs) across the South American Camelids (SACs) genome to identify the genetic relationship between the four species and the potential signatures of selection. RESULTS: A total of 37 WGS samples covering the four species was included in the final analysis. The multi-dimensional scaling approach showed a clear separation between the four species; however, admixture analysis suggested a strong genetic introgression from vicugna and llama to alpaca. Conversely, very low genetic admixture of the guanaco with the other SACs was found. The four species did not show significant differences in the number, length of ROHs (100-500 kb) and genomic inbreeding values. Longer ROHs (> 500 kb) were found almost exclusively in alpaca. Seven overlapping ROHs were shared by alpacas, encompassing nine loci (FGF5, LOC107034918, PRDM8, ANTXR2, LOC102534792, BSN, LOC116284892, DAG1 and RIC8B) while nine overlapping ROHs were found in llama with twenty-five loci annotated (ERC2, FZD9, BAZ1B, BCL7B, LOC116284208, TBL2, MLXIPL, PHF20, TRNAD-AUC, LOC116284365, RBM39, ARFGEF2, DCAF5, EXD2, HSPB11, LRRC42, LDLRAD1, TMEM59, LOC107033213, TCEANC2, LOC102545169, LOC116278408, SMIM15, NDUFAF2 and RCOR1). Four overlapping ROHs, with three annotated loci (DLG1, KAT6B and PDE4D) and three overlapping ROHs, with seven annotated genes (ATP6V1E1, BCL2L13, LOC116276952, BID, KAT6B, LOC116282667 and LOC107034552), were detected for vicugna and guanaco, respectively. CONCLUSIONS: The signatures of selection revealed genomic areas potentially selected for production traits as well as for natural adaptation to harsh environment. Alpaca and llama hint a selection driven by environment as well as by farming purpose while vicugna and guanaco showed selection signals for adaptation to harsh environment. Interesting, signatures of selection on KAT6B gene were identified for both vicugna and guanaco, suggesting a positive effect on wild populations fitness. Such information may be of interest to further ecological and animal production studies.
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Camelídeos Americanos , Animais , Humanos , Camelídeos Americanos/genética , Grupos Raciais , Aclimatação , Agricultura , América do Sul , Receptores de Peptídeos , Fatores de Transcrição , Histona AcetiltransferasesRESUMO
Middle East respiratory syndrome coronavirus (MERS-CoV) clade B viruses are found in camelids and humans in the Middle East, but clade C viruses are not. We provide experimental evidence for extended shedding of MERS-CoV clade B viruses in llamas, which might explain why they outcompete clade C strains in the Arabian Peninsula.
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Camelídeos Americanos , Infecções por Coronavirus , Herpesvirus Cercopitecino 1 , Coronavírus da Síndrome Respiratória do Oriente Médio , Animais , Humanos , Eliminação de Partículas Virais , CamelusRESUMO
Hormonally active tumours are characterized by production and secretion of hormones, irrespective of endogenous feedback mechanisms. An adult llama had exuberant oestrous behaviour, infertility, elevated concentrations of oestradiol and a large ovarian mass. Necropsy revealed the presence of two large abdominal masses, one effacing the right ovary and one in the mesocolon. Considering the clinical and histopathological findings, we conclude that the llama was affected by a granulosa cell tumour. The case suggests that granulosa cell tumours in camelids are hormonally active, and the clinical presentation resembles that of other large animal species. To our knowledge, this is the first case report of an oestrogen-producing, metastatic granulosa cell tumour in a llama.
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Camelídeos Americanos , Tumor de Células da Granulosa , Neoplasias Ovarianas , Feminino , Animais , Tumor de Células da Granulosa/veterinária , Neoplasias Ovarianas/veterináriaRESUMO
There are no available data regarding the hematology, serum biochemistry, and fore stomach fluid constituents of llama (Lama glama) in Egypt. This study aimed to establish normal reference values for blood and fore stomach fluid constituents of llama and determine the influence of sex and season on these parameters under Egyptian conditions. The study was performed on (n = 38; 22 female, 16 male; 1-7 years) apparently healthy llamas located in the Giza Zoo and private zoo in the Ismailia Governorate. Samples were collected in two seasons and divided into summer and winter samples. Differences in the mean and range values of packed cell volume, serum minerals, fore stomach fluid pH, and total protozoal count in Egypt were recorded. Sex and season had minimal effects on hematology and only erythrocyte count showed a significant (p < 0.05) increase in males compared with females. Regarding serum biochemistry, males showed significant (p < 0.05) increases in alanine transaminase and calcium levels, while globulin significantly (p < 0.05) increased in females. The influence of season on serum biochemistry was evident in alanine transaminase, total protein, albumin, and chloride which increased significantly (p < 0.05) in summer, while urea, bilirubin, and magnesium increased significantly (p < 0.05) in winter. Fore stomach fluid pH and ammonia showed significant (p < 0.05) increases in winter, while the total protozoal count increased significantly (p < 0.05) in summer and in males compared with females. The results obtained in this study can serve as reference values for the hematobiochemical and fore stomach fluid constituents of llama in Egypt.
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Camelídeos Americanos , Feminino , Masculino , Animais , Egito , Alanina Transaminase , Contagem de Eritrócitos/veterinária , CamelidaeRESUMO
Natural language processing (NLP) has been studied in computing for decades. Recent technological advancements have led to the development of sophisticated artificial intelligence (AI) models, such as Chat Generative Pre-trained Transformer (ChatGPT). These models can perform a range of language tasks and generate human-like responses, which offers exciting prospects for academic efficiency. This manuscript aims at (i) exploring the potential benefits and threats of ChatGPT and other NLP technologies in academic writing and research publications; (ii) highlights the ethical considerations involved in using these tools, and (iii) consider the impact they may have on the authenticity and credibility of academic work. This study involved a literature review of relevant scholarly articles published in peer-reviewed journals indexed in Scopus as quartile 1. The search used keywords such as "ChatGPT," "AI-generated text," "academic writing," and "natural language processing." The analysis was carried out using a quasi-qualitative approach, which involved reading and critically evaluating the sources and identifying relevant data to support the research questions. The study found that ChatGPT and other NLP technologies have the potential to enhance academic writing and research efficiency. However, their use also raises concerns about the impact on the authenticity and credibility of academic work. The study highlights the need for comprehensive discussions on the potential use, threats, and limitations of these tools, emphasizing the importance of ethical and academic principles, with human intelligence and critical thinking at the forefront of the research process. This study highlights the need for comprehensive debates and ethical considerations involved in their use. The study also recommends that academics exercise caution when using these tools and ensure transparency in their use, emphasizing the importance of human intelligence and critical thinking in academic work.
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The objective of the study was to characterize the anatomical framework and sites of action of the nerve growth factor (NGF)-mediated ovulation-inducing system of llamas. The expression patterns of NGF and its receptors in the hypothalamus of llamas (n = 5) were examined using single and double immunohistochemistry/immunofluorescence. We also compare the expression pattern of the P75 receptor in the hypothalamus of llama and a spontaneous ovulator species (sheep, n = 5). Both NGF receptors (TrkA and P75) were highly expressed in the medial septum and diagonal band of Broca, and populations of TrkA cells were observed in the periventricular and dorsal hypothalamus. Unexpectedly, we found NGF immunoreactive cell bodies with widespread distribution in the hypothalamus but not in areas endowed with NGF receptors. The organum vasculosum of the lamina terminalis (OVLT) and the median eminence displayed immunoreactivity for P75. Double immunofluorescence using vimentin, a marker of tanycytes, confirmed that tanycytes were immunoreactive to P75 in the median eminence and in the OVLT. Additionally, tanycytes were in close association with GnRH and kisspeptin in the arcuate nucleus and median eminence of llamas. The choroid plexus of llamas contained TrkA and NGF immunoreactivity but no P75 immunoreactivity. Results of the present study demonstrate sites of action of NGF in the llama hypothalamus, providing support for the hypothesis of a central effect of NGF in the ovulation-inducing mechanism in llamas.
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Hipotálamo/fisiologia , Fator de Crescimento Neural/metabolismo , Ovulação/fisiologia , Receptor de Fator de Crescimento Neural/metabolismo , Animais , Camelídeos Americanos , Plexo Corióideo , Feminino , Imunofluorescência , Regulação da Expressão Gênica , Hormônio Liberador de Gonadotropina/genética , Hormônio Liberador de Gonadotropina/metabolismo , Imuno-Histoquímica , Kisspeptinas/genética , Kisspeptinas/metabolismo , Fator de Crescimento Neural/genética , Receptor de Fator de Crescimento Neural/genética , Receptor trkA/genética , Receptor trkA/metabolismo , Ovinos/fisiologia , Vimentina/genética , Vimentina/metabolismoRESUMO
Semen cryopreservation is not available for massive use in South American Camelids (SACs) due to the lack of an efficient protocol and the low pregnancy rates obtained with artificial insemination (AI). The use of a single cryoprotectant (CP) is commonly used in SACs frozen semen. The objective of the study was to evaluate the combined cryoprotective capacity of two permeable CPs at different stages of the cryopreservation protocol in llama semen. Sixteen ejaculates from 4 llama males were analysed, and sperm quality was assayed in raw semen, at 5°C, after equilibration of samples with the CPs and when samples were thawed. The following CPs and combination were used: 6% glycerol (GL), 6% dimethylformamide (DMF) and the combination of both CPs: 3% GL and 3% DMF. A Kruskal-Wallis test and an experimental factorial design, considering one factor with four levels (raw semen, 6% GL, 6% DMF and GL/DMF), were used. Total sperm motility and live sperm with intact acrosomes remained unchanged after equilibration of samples (p > .05). A significant decrease in the percentage of functional membrane, motile and live sperm with intact acrosomes was observed when samples were thawed (GL, DMF and GL/DMF). Nevertheless, the cryopreservation protocols used preserved sperm DNA quality; thus, sperm chromatin condensation and DNA fragmentation were unaffected (p > .05) when GL, DMF and GL/DMF were used. To conclude, no superiority was found between the use of a single or a combination of permeable cryoprotectants to freeze llama semen.
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Camelídeos Americanos , Criopreservação/veterinária , Crioprotetores/farmacologia , Espermatozoides/efeitos dos fármacos , Acrossomo , Animais , Criopreservação/métodos , Fragmentação do DNA , Dimetilformamida/farmacologia , Glicerol/farmacologia , Masculino , Análise do Sêmen/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacosRESUMO
Given the predisposition of South American camelids to coccidioidomycosis, we sought to describe the disease presentation in alpacas and llamas and identify potential risk factors for these species. The records of 224 llamas and alpacas that were tested for Coccidioides infection using immunodiffusion serology at the Coccidioidomycosis Serology Laboratory of the University of California, Davis, between 1990 and 2016 were examined; of those, 46 alpacas and 42 llamas had positive test results. The remaining 99 alpacas and 37 llamas were used as control groups. We found that male llamas were at increased risk for Coccidioides infection when compared with female llamas and when compared with male alpacas. South American camelids living within California were at higher risk for infection than camelids living in other states. Alpacas were more likely than llamas to have subclinical infections. We documented five cases of abortion or neonatal mortality attributable to coccidioidomycosis in alpacas. Our study demonstrates that South American camelids are susceptible to Coccidioides infection in areas where the disease is endemic, lending support to the importance of vigilance for this disease in alpacas and llamas and suggesting a possible role for these animals as sentinel species. LAY SUMMARY: We examined cases of Valley Fever and described the disease and risk factors for llamas and alpacas. Male llamas were at increased risk for infection as were animals living within California. Five alpacas had miscarriages or neonatal deaths as a result of Valley Fever infections.
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The aim of this study was to characterize corpus luteum vascularization and its association with plasma progesterone concentration in early stages of pregnancy, when maternal recognition of pregnancy is expected to occur. In all animals, both plasma progesterone concentration and corpus luteum vascularization increased from Day 6 to Day 8 post-mating and afterwards in non-pregnant llamas they started to decrease to reach basal levels around Days 12 to 14 post-mating, while in pregnant animals, both variables remained elevated until the end of the study. A lineal positive relationship between corpus luteum vascularization and plasma progesterone concentration was observed in pregnant (r2 = .46, p < .0001) and non-pregnant llamas (r2 = .66, p < .0001). Pregnant animals showed higher plasma progesterone concentration and corpus luteum vascularization than the non-pregnant ones from Day 12 post-mating until the end of the study (p Ë .05 and p Ë .01, respectively). These results suggest that maternal recognition of pregnancy should occur before Day 12 post-mating in order to expand luteal lifespan, maintaining corpus luteum vascularization and progesterone production. Also, the assessment of CL vascularization area could be a useful and non-invasive method for early pregnancy diagnosis due to its association with plasma progesterone concentration.
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Camelídeos Americanos/fisiologia , Corpo Lúteo/irrigação sanguínea , Progesterona/sangue , Animais , Corpo Lúteo/fisiologia , Feminino , Masculino , Ovário/diagnóstico por imagem , Gravidez , Ultrassonografia/veterináriaRESUMO
Camelid ungulates produce homodimeric heavy chain-only antibodies (HCAbs) in addition to conventional antibodies consisting of paired heavy and light chains. In the llama, HCAbs are made up by at least two subclasses (long-hinge IgG2b and short-hinge IgG2c HCAbs vs. conventional heterotetrameric IgG1s). Here, we generated murine monoclonal antibodies (mAbs) specific for the hinge-CH2 boundary of llama IgG2b (mAb 1C10) and the Fc of llama IgG2c HCAbs (mAb 5E4). Flow cytometric analysis of llama peripheral blood lymphocytes revealed that IgG1+, IgG2b+ and IgG2c+ B cells could be distinguished using mAbs 1C10/5E4 but had equivalent expression of three other cell-surface markers. MiSeq sequencing of the peripheral B cell repertoires of three llamas showed that (i) IgG2b and IgG2c HCAbs were present in similar proportions in the repertoire, (ii) a subset of IgG2b and IgG2c HCAbs, but not IgG1s, entirely lacked a hinge exon and showed direct VHH-CH2 splicing; these "hingeless" HCAbs were clonally expanded, somatically mutated and derived from hinged HCAb precursors, (iii) substantial repertoire overlap existed between IgG subclasses, especially between IgG2b and IgG2c HCAbs, (iv) the complementarity-determining region (CDR)-H3 length distributions of IgG2b and IgG2c HCAbs were broader and biased towards longer lengths compared with IgG1s due to increased N-nucleotide addition, (v) IgG2b and IgG2c HCAbs used a more restricted set of IGHV genes compared with IgG1s, and (vi) IgG2b and IgG2c HCAbs had elevated somatic mutations rates of both CDRs and framework regions (FRs) compared with IgG1s, especially of CDR-H1 and FR3. The distinct molecular features of llama IgG1, IgG2b and IgG2c antibodies imply that these subclasses may have divergent immunological functions and suggest that specific mechanisms operate to diversify HCAb repertoires in the absence of a light chain.
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Anticorpos Monoclonais/imunologia , Linfócitos B/imunologia , Camelídeos Americanos/imunologia , Regiões Determinantes de Complementaridade/imunologia , Imunoglobulina G/imunologia , Cadeias Pesadas de Imunoglobulinas/imunologia , Animais , Linfócitos B/metabolismo , Camelídeos Americanos/genética , Regiões Determinantes de Complementaridade/genética , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala , Imunogenética/métodos , Imunoglobulina G/genética , Cadeias Pesadas de Imunoglobulinas/genética , Imunofenotipagem/métodos , CamundongosRESUMO
BACKGROUND: Nerve growth factor (ß-NGF) from llama seminal plasma has been described as a potent ovulatory and luteotrophic molecule after intramuscular or intrauterine infusion in llamas and alpacas. We tested the hypothesis that systemic administration of purified ß-Nerve Growth Factor (ß-NGF) during the preovulatory stage will up-regulate steroidogenic enzymes and Vascular Endothelial Growth Factor (VEGF) gene expression in granulosa cells inducing a change in the progesterone/estradiol ratio in the follicular fluid in llamas. METHODS: Experiment I: Female llamas (n = 64) were randomly assigned to receive an intramuscular administration of: a) 50 µg gonadorelin acetate (GnRH, Ovalyse, Pfizer Chile SA, Santiago, Chile, n = 16), b) 1.0 mg of purified llama ß-NGF (n = 16), or c) 1 ml phosphate buffered saline (PBS, negative control group, n = 16). An additional group of llamas (n = 16) were mated with a fertile male. Follicular fluid and granulosa cells were collected from the preovulatory follicle at 10 or 20 h after treatment (Time 0 = administration of treatment, n = 8/treatment/time point) to determine progesterone/estradiol concentration and steroidogenic enzymes and VEGF gene expression at both time points. Experiment II: Granulosa cells were collected from preovulatory follicles from llamas (n = 24) using ultrasound-guided transvaginal follicle aspiration for in vitro culture to determine mRNA relative expression of Steroidogenic Acute Regulatory Protein (StAR) and VEGF at 10 or 20 h (n = 4 replicates) and progesterone secretion at 48 h (n = 4 replicates) after LH or ß-NGF treatment. RESULTS: Experiment I: There was a significant increase in the progesterone/estradiol ratio in mated llamas or treated with GnRH or purified ß-NGF. There was a significant downregulation in the mRNA expression of Aromatase (CYP19A1/P450 Arom) for both time points in llamas mated or treated with GnRH or llama purified ß-NGF with respect to the control group. All treatments except ß-NGF (20 h) significantly up regulated the mRNA expression of 3-beta-hydroxysteroid dehydrogenase (HSD3B) whereas the expression of StAR and Side-Chain cleavage enzyme (CYP11A1/P450scc) where significantly up regulated only by mating (20 h), or ß-NGF at 10 or 20 h after treatment. VEGF was up regulated only in those llamas submitted to mating (10 h) or treated with purified ß-NGF (10 and 20 h). Experiment II: Only ß-NGF treatment induced an increase of mRNA abundance of StAR from llama granulosa cells at 20 h of in vitro culture. There was a significant increase on mRNA abundance of VEGF at 10 and 20 h of in vitro culture from granulosa cells treated with ß-NGF whereas LH treatment increases VEGF mRNA abundance only at 20 h of in vitro culture. In addition, there was a significant increase on progesterone secretion from llama granulosa cells 48 h after LH or ß-NGF treatment. CONCLUSIONS: Systemic administration of purified ß-NGF from llama seminal fluid induced a rapid shift from estradiol to progesterone production in the preovulatory follicle. Differences in gene expression patterns of steroidogenic enzymes between GnRH and mated or ß-NGF-treated llamas suggest local effects of seminal components on the preovulatory follicle.
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Camelídeos Americanos/fisiologia , Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , Fator de Crescimento Neural/farmacologia , Sêmen/química , Animais , Estradiol/sangue , Feminino , Perfilação da Expressão Gênica , Fosfoproteínas/metabolismo , Progesterona/sangue , RNA Mensageiro/metabolismo , Distribuição Aleatória , Reprodução/fisiologia , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
In South American camelids, sperm survival is low after thawing and poor results are obtained when artificial insemination is performed with cryopreserved semen. The aim of this study was to evaluate the effect of different percentages (10% and 50%) of seminal plasma added prior to the process of cryopreservation and also to evaluate the absence of seminal plasma on llama sperm survival after freezing and thawing. A total of 15 ejaculates from five adult llama males (n = 5; r = 3) were evaluated. A significant decrease in sperm motility, viability, membrane function and intact acrosomes was observed in thawed samples (0%, 10% and 50%) when compared to raw semen. Neither morphology nor chromatin condensation was altered in all thawed samples (p > 0.05), but a significant increase (p < 0.05) in the percentage of spermatozoa with fragmented DNA was observed after thawing all samples compared to raw semen. Higher percentages of total and progressive sperm motility were observed when 0% and 10% of seminal plasma were used compared to 50%. However, no statistical differences were established for sperm viability, membrane function, morphology, acrosome status and DNA quality between thawed treatments. To conclude, neither of the percentages of seminal plasma used showed superiority nor cryoprotective effect on llama sperm survival.
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Camelídeos Americanos , Criopreservação/veterinária , Crioprotetores/farmacologia , Preservação do Sêmen/métodos , Sêmen , Animais , Sobrevivência Celular/efeitos dos fármacos , Inseminação Artificial/métodos , Inseminação Artificial/veterinária , Masculino , Análise do Sêmen , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacosRESUMO
The objectives of this study were as follows: (i) to describe and evaluate the frequencies of different morphologies of llama sperm nuclei, (ii) to determine morphometric values of nuclear parameters, (iii) to describe and estimate the frequencies of different classes of chromatin distribution and (iv) to measure haploid DNA content and analyse its nuclear distribution. The study was performed using ejaculates collected from seven males, and sperm nuclei were stained with the Feulgen reaction. Normal morphology ranged from 78.36% to 93.92%, and abnormalities included short, small, large, pyriform, narrow, micro and round nuclei. Important differences in nuclei considered normal were found between some males. The following average values were obtained for each sperm nuclear morphometric parameter analysed: area 11.64 µm2 , perimeter 13.16 µm, length 5.12 µm, width 2.81 µm, ellipticity 1.85 and form 0.83. Differences between males were significant for all the parameters (p < .01). Light microscope observations and cytophotometric determinations allowed discriminating between three classes of chromatin distribution: homogeneous, diffuse and showing a clear band. Significant differences between males were found for the frequencies of the three classes (p < .01). Cluster analysis methods were used to estimate the resemblance between males according to the characteristics of their sperm nuclei. A great intermale variability was found for morphological, morphometric and chromatin distribution data. These parameters would have low dependence between them.
Assuntos
Camelídeos Americanos , Núcleo Celular/ultraestrutura , Espermatozoides/citologia , Animais , Cromatina , Análise por Conglomerados , MasculinoRESUMO
The study objective was to evaluate the effects of age on aminoglycoside pharmacokinetics in eight young-adult (<4 years) and eight aged (≥14 years) healthy alpacas, receiving a single 6.6 mg/kg intravenous gentamicin injection. Heparinized plasma samples were obtained at designated time points following drug administration and frozen at -80°C until assayed by a validated immunoassay (QMS® ). Compartmental and noncompartmental analyses of gentamicin plasma concentrations versus time were performed using WinNonlin (v6.4) software. Baseline physical and hematological parameters were not significantly different between young and old animals with the exception of sex. Data were best fitted to a two-compartment pharmacokinetic model. The peak drug concentration at 30 min after dosing (23.8 ± 2.1 vs. 26.1 ± 2 µg/ml, p = .043) and area under the curve (70.4 ± 10.5 vs. 90.4 ± 17.6 µg hr/ml, p = .015) were significantly lower in young-adult compared to aged alpacas. Accordingly, young alpacas had a significantly greater systemic clearance than older animals (95.5 ± 14.4 and 75.6 ± 16.1 ml hr-1 kg-1 ; p = .018), respectively). In conclusion, a single 6.6 mg/kg intravenous gentamicin injection achieves target blood concentrations of >10 times the MIC of gentamicin-susceptible pathogens with MIC levels ≤2 µg/ml, in both young-adult and geriatric alpacas. However, the observed reduction in gentamicin clearance in aged alpacas may increase their risk for gentamicin-related adverse drug reactions.
Assuntos
Antibacterianos/farmacocinética , Camelídeos Americanos/metabolismo , Gentamicinas/farmacocinética , Fatores Etários , Animais , Antibacterianos/administração & dosagem , Antibacterianos/sangue , Camelídeos Americanos/sangue , Feminino , Gentamicinas/administração & dosagem , Gentamicinas/sangue , Meia-Vida , Injeções Intravenosas/veterinária , MasculinoRESUMO
Middle East respiratory syndrome (MERS) cases continue to be reported, predominantly in Saudi Arabia and occasionally other countries. Although dromedaries are the main reservoir, other animal species might be susceptible to MERS coronavirus (MERS-CoV) infection and potentially serve as reservoirs. To determine whether other animals are potential reservoirs, we inoculated MERS-CoV into llamas, pigs, sheep, and horses and collected nasal and rectal swab samples at various times. The presence of MERS-CoV in the nose of pigs and llamas was confirmed by PCR, titration of infectious virus, immunohistochemistry, and in situ hybridization; seroconversion was detected in animals of both species. Conversely, in sheep and horses, virus-specific antibodies did not develop and no evidence of viral replication in the upper respiratory tract was found. These results prove the susceptibility of llamas and pigs to MERS-CoV infection. Thus, the possibility of MERS-CoV circulation in animals other than dromedaries, such as llamas and pigs, is not negligible.