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OBJECTIVES: Chemoprevention can be a treatment for potentially malignant lesions (PMLs). We aimed to evaluate whether artemisinin (ART) and cisplatin (CSP) are associated with apoptosis and immunogenic cell death (ICD) in vitro, using oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC) cell lines, and whether these compounds prevent OL progression in vivo. METHODS: Normal keratinocytes (HaCat), Dysplastic oral cells (DOK), and oral squamous cell carcinoma (SCC-180) cell lines were treated with ART, CSP, and ART + CSP to analyze cytotoxicity, genotoxicity, cell migration, and increased expression of proteins related to apoptosis and ICD. Additionally, 41 mice were induced with OL using 4NQO, treated with ART and CSP, and their tongues were histologically analyzed. RESULTS: In vitro, CSP and CSP + ART showed dose-dependent cytotoxicity and reduced SCC-180 migration. No treatment was genotoxic, and none induced expression of proteins related to apoptosis and ICD; CSP considerably reduced High-mobility group box-1 (HMGB-1) protein expression in SCC-180. In vivo, there was a delay in OL progression with ART and CSP treatment; however, by the 16th week, only CSP prevented progression to OSCC. CONCLUSION: Expression of proteins related to ICD and apoptosis did not increase with treatments, and CSP was shown to reduce immunogenic pathways in SCC-180, while reducing cell migration. ART did not prevent the malignant progression of OL in vivo; CSP did despite significant adverse effects.

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Integrating in vitro embryo production with embryonic microsurgery facilitates the generation of monozygotic twins. However, despite their potential benefits, these methods have not been widely adopted in commercial settings because of their substantial costs. Hence, there is a need to streamline the bisection procedure while ensuring efficient production of viable demi-embryos. In this study, we investigated the impact of different orientations of microsurgical incisions in relation to inner cell mass on embryonic development, morphology, viability, and expression of cell fate protein markers using a simplified microsurgery approach. Ovaries were transported from the slaughterhouse to the laboratory and aspirated to obtain oocytes that were selected and subjected to in vitro embryo production. The selected expanded blastocysts (n = 204) underwent microsurgery. The blastocysts were immobilized to facilitate incision using an adapted microblade, yielding demi-embryos (vertical incision) and viable embryonic fragments (transverse incision). The structures were then re-cultured for 12 h. Viability was assessed by measuring the re-expansion rate after re-culture, followed by immunofluorescence analysis of proteins (CDX2 and NANOG) and apoptosis analysis using terminal deoxynucleotyl transferase dUTP nick end-labeling (TUNEL). Microsurgically derived embryos exhibited remarkable plasticity, as evidenced by a slight reduction (P < 0.05) in the re-expansion rate (transverse 64.2 % and vertical 57.2 %) compared to that of the control group (blastocysts without microsurgery) (86.7 %). They also demonstrated the ability of morphological reconstitution after culturing. Despite the anticipated decrease (P < 0.05) in the total number of cells and embryo volume, microsurgery did not result in a significant increase (P > 0.05) in the number of apoptotic cells. Furthermore, microsurgery led to higher (P < 0.05) expression of markers associated with pluripotency, indicating its efficiency in preserving regenerative capacity. Moreover, microsurgery, whether followed by immunosurgery or not, made the isolation of embryonic cells easier. In conclusion, both transverse and vertical microsurgery incisions enabled the production of identical demi-embryos and served as tools for isolating embryonic cells without compromising the resumption of development and the apoptotic index.

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Intravascular papillary endothelial hyperplasia (IPEH) represents an uncommon reactive endothelial hyperplastic proliferation. A 46-year-old man experienced increased volume in the right maxilla, elevation of the nasal ala, and swelling of the hard palate with a reddish hue for 3 months. Computed tomography revealed an expansive hypodense region and cortical bone destruction associated with an impacted supernumerary tooth and an endodontically treated tooth. Under the differential diagnoses of a radicular cyst, dentigerous cyst, and ameloblastoma, an exploratory aspiration and incisional biopsy were performed. This revealed the formation of blood vessels of various diameters lined by endothelium, forming intravascular papillae positive for CD-34. The definitive diagnosis was IPEH, and the patient was treated by embolization and surgery. Histological analysis confirmed the presence of IPEH associated with an odontogenic cyst. After 12 months of follow-up, no recurrence was observed. Also, we reviewed case reports of IPEH affecting the maxilla and mandible. Fourteen intraosseous cases were reported in the maxilla and mandible, with a preference for males and affecting a wide age range. Complete surgical excision was the treatment of choice, and recurrences were not reported. The pathogenesis of IPEH is controversial and may originate from trauma or inflammatory processes. To the best of our knowledge, this is the first report of an association of IPEH with an odontogenic cyst. The importance of IPEH in the differential diagnosis of intraosseous lesions in the jaws is emphasized, and preoperative semiotic maneuvers are needed to prevent surgical complications.

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Several variables can influence the treatment and outcome of orthognathic surgery, one of which is the angle of the mandibular ramus. The objective of this study is to analyze the upper airways (UA) of patients prior to orthognathic surgery, using cone-beam computed tomography (CBCT) and to correlate their volume with the angle of the mandibular ramus. DICOM (Digital Image Communication in Medicine) images of a radiology clinic were used, with a survey of 124 full face CBCT scans, from 2015 to 2018, and the measurement of the total volume in mm³ through software. To obtain the values of the angle of the mandibular ramus, images obtained from medical reports were used, through morphological evaluation. The values were tabulated and subsequently statistical analysis was performed using the ANOVA test to assess the differences between the mean airway volumes (mm³) according to gender, age and mandibular ramus angle. However, the angle of the ramus did not influence the volume of the UA, but a greater average of UA volume was observed in male individuals and in individuals over 34 years of age. There is no evidence that it is possible to measure or predict the volume of the UA by evaluating the angle of the mandibular ramus. (AU)

Diversas as variáveis podem influenciar no tratamento e resultado de uma cirurgia ortognática, uma delas é o ângulo do ramo mandibular. O objetivo deste trabalho é analisar as vias aéreas superiores (VAS) de pacientes previamente à cirurgia ortognática, por meio de tomografia computadorizada de feixe cônico (TCFC) e correlacionar o seu volume com o ângulo do ramo mandibular. Foram utilizadas imagens DICOM (Comunicação de Imagens Digitais em Medicina) de uma clínica radiológica, com o levantamento de 124 exames de TCFC de face total, do ano de 2015 até 2018 e feita a mensuração do volume total em mm³ através de um software. Para a obtenção dos valores do ângulo do ramo mandibular foram utilizadas imagens obtidas através dos laudos médicos, mediante avaliação morfológica. Os valores foram tabulados e posteriormente feita a análise estatística com o teste ANOVA para avaliar as diferenças entre as médias de volume das vias aéreas (mm³) de acordo com o gênero, a idade e o ângulo do ramo mandibular. Contudo, o ângulo do ramo não influenciou no volume das VAS, mas uma maior média de volume de VAS foi observada nos indivíduos do gênero masculino e nos indivíduos acima de 34 anos. Não há evidências de que se é possível mensurar ou ter previsibilidade do volume de VAS, avaliando o ângulo do ramo mandibular. (AU)

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The beneficial effects of high-fat low-carbohydrate (HFLC) diets on glucose metabolism have been questioned and their effects on liver metabolism are not totally clear. The aim of this work was to investigate the effects of an HFLC diet under different energy conditions on glucose homeostasis, fatty liver development, and hepatic gluconeogenesis using the isolated perfused rat liver. HFLC diet (79% fat, 19% protein, and 2% carbohydrates in Kcal%) was administered to rats for 4 weeks under three conditions: ad libitum (hypercaloric), isocaloric, and hypocaloric (energy reduction of 20%). Fasting blood glucose levels and total fat in the liver were higher in all HFLC diet rats. Oral glucose tolerance was impaired in isocaloric and hypercaloric groups, although insulin sensitivity was not altered. HFLC diet also caused marked liver metabolic alterations: higher gluconeogenesis rate from lactate and a reduced capacity to metabolize alanine, the latter effect being more intense in the hypocaloric condition. Thus, even when HFLC diets are used for weight loss, our data imply that they can potentially cause harmful consequences for the liver.

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In this study, we investigated the impact of incorporating Interleukin-13 (IL-13) into the embryonic culture medium and its influence on cryotolerance and cellular viability of vitrified bovine embryos. Two distinct time points for IL-13 supplementation were explored: during the final hours of culture prior to cryopreservation and during the period of recultivation following cryopreservation and warming. Cryosurvival rates, total cell count, and cell viability were assessed using the TUNEL technique to determine the apoptotic percentage. Re-expansion and hatching rates did not show differences among all groups (P > 0.05), and the total cell number was comparable between the treated and control groups (P > 0.05). However, the group that received IL-13 before vitrification exhibited a higher apoptotic percentage (P < 0.05). This suggests that the anti-inflammatory effect of IL-13 may have impacted the embryo's defense capacity against the stress induced by cryopreservation, leading to an increased percentage of apoptosis, although it did not influence the developmental resumption capability.

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In vitro-produced embryos are constantly exposed to stressful conditions that can lead to the activation of the apoptotic pathway. The nuclear Kappa B factor (NF-κB) is an inflammatory mediator that induces the expression of tumor necrosis factor (TNF-α), a pro-inflammatory cytokine, while interleukin-10 (IL-10), an anti-inflammatory cytokine, inhibits NF-κB activity. This study aimed to investigate the effects of IL-10 and TNF-α on the competence and cryosurvival of in vitro-produced bovine embryos. Embryos were produced in vitro using standard protocols, and Grade I blastocysts were vitrified using the Cryotop method. Non-vitrified and vitrified blastocysts were subjected to the TUNEL assay. In Experiment I, on day 6.5 (156 h post-insemination), the embryos were treated with PBS (control), 50 ng/mL of IL-10, or a combination of 25 ng/mL of TNF-α and 50 ng/mL of IL-10. Embryonic development and apoptotic rates were monitored. In Experiment II, the same groups were set up, with the addition of a group treated with 25 ng/mL of TNF-α alone. Grade I blastocysts were vitrified 5 h after treatment, and cryosurvival was monitored at until 48 h post-warming. The apoptosis rate and total cell number were investigated in the vitrified-hatched blastocysts. IL-10 alone did not affect developmental competence or cryosurvival (P > 0.05). The IL-10-treated embryos, when exposed in combination with TNF-α, presented a detrimental effect (P < 0.05) in the embryonic development of non-vitrified embryos. However, vitrified blastocysts had no negative effect (P > 0.05). The TNF-α treatment reduced (P < 0.05) the re-expansion rate at 6 h post-warming and increased (P < 0.05) the apoptosis rate in vitrified hatched blastocysts, whereas no effect (P > 0.05) of the treatments was detected in the hatching rate and total cell number post-warming. In conclusion, TNF-α has a detrimental effect on embryonic developmental competence and cryosurvival by compromising the development of non-vitrified embryos and apoptotic-related events of vitrified blastocysts, whereas IL-10, when in combination with TNF-α, appears to attenuate the detrimental effects of TNF-α.

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Presentamos el caso de un hombre de 45 años con úlcera en paladar de 10 días de evolución. El paciente era consumidor de alcohol y tenía antecedentes de consumo de cocaína desde hacía 17 años, discontinuada durante los últimos 7 años. Se realizó biopsia incisional y los cortes histológicos revelaron una extensa área ulcerada recubierta por una membrana fibrinopurulenta, con pequeños nidos de células escamosas que presentaban citoplasma eosinófilo, núcleos evidentes y en ocasiones múltiples, leve atipia celular y varias figuras mitóticas. Las características clínicas e histológicas indicaron el diagnostico de Sialometaplasia Necrotizante y el paciente fue sometido a seguimiento clínico. En la tercera semana de seguimiento se hizo evidente la remisión de la lesión. Destacamos la importancia del examen histopatológico e inmunohistoquímico en el correcto diagnóstico.

Relatamos o caso de um homem de 45 anos com úlcera no palato há 10 dias. O paciente era usuário de álcool e tinha história de uso de cocaína há 17 anos, interrompida há 7 anos. Foi realizada biópsia incisional e os cortes histológicos revelaram extensa área ulcerada recoberta por membrana fibrinopurulenta, com pequenos ninhos de células escamosas que apresentavam citoplasma eosinofílico, núcleos evidentes e por vezes múltiplos, atipias celulares leves e diversas figuras mitóticas. As características clínicas e histológicas indicaram o diagnóstico de Sialometaplasia Necrotizante e o paciente foi submetido a acompanhamento clínico. Na terceira semana de acompanhamento, a remissão da lesão tornou-se evidente. Ressaltamos a importância do exame histopatológico e imunohistoquímico no correto diagnóstico.

We report a case of a 45-year-old man with a palate ulcer for 10 days. The patient was an alcohol user and had a history of using cocaine for 17 years, discontinued for the last 7 years. An incisional biopsy was performed, and histological sections revealed an extensive ulcerated area covered by a fibrinopurulent membrane, with small nests of squamous cells in the connective tissue. These squamous cells exhibited eosinophilic cytoplasm, evident and sometimes multiple nuclei, mild cellular atypia, and several mitotic figures. The clinical and histological characteristics indicated a Necrotizing Sialometaplasia diagnosis, and the patient underwent clinical follow-up. In the third week of follow-up, remission of the lesion became apparent. We emphasize the importance of histopathological and immunohistochemical examination in the correct diagnosis.

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Novos fármacos, como a artemisinina (ART), podem ser promissores no tratamento de lesões potencialmente malignas (LPM) e podem ser úteis quando usados em associação com outros quimioterápicos, especialmente na redução dos seus efeitos colaterais. A leucoplasia oral (LO) é a LPM mais comum da cavidade bucal e, pode evoluir para um carcinoma de células escamosas oral (CCEO). Não há terapia para evitar a sua transformação maligna, a quimioprevenção pode iniciar a morte celular imunogênica (MCI) que ativa o sistema imunológico para que reconheça e elimine as células malignas ou pré-malignas, sendo um potencial tratamento para as LPM. O presente estudo objetivou avaliar se a ART e a cisplatina (CSP) associadas ou não seriam capazes de induzir a MCI em linhagens celulares de LO (DOK) e CCEO SCC180). Material e métodos: As linhagens celulares HaCat (controle), DOK e SCC-180 foram tratadas por ART e CSP de forma combinada ou isolada, a fim de analisar a citotoxicidade e a genotoxicidade destes fármacos, além da capacidade destes em reduzir a migração celular e, se os compostos seriam capazes de induzir a expressão da proteína box de alta mobilidade (HGMB-1), caspase 3, 8, 9, e Calreticulina (CALR). Resultados: Em todas as linhagens celulares a CSP e CSP+ART causaram uma resposta dose dependente, apresentando maior citotoxicidade com doses mais altas, o que não foi observado com a ART. A formação de micronúcleos não foi observada no teste de genotoxicidade. A taxa de migração foi reduzida com as concentrações de IC50 de CSP e ART+CSP para as células de CCEO. Não foram encontradas expressões significativas de proteínas relacionadas à MCI ou apoptose nas linhagens de LO e CCEO, tratadas com ART, CSP ou ART+CSP, indicando que outro tipo de morte celular possa ter ocorrido. Conclusão: A MCI e a apoptose não foram evidenciadas como forma de morte celular após as linhagens de LO e CCEO receberem tratamentos com ART, CSP e a associação de ambas. Efeitos genotóxicos não foram observados nas doses testadas. O tratamento de CSP e ART+CSP foi capaz de reduzir a migração de células de CCEO. Também concluímos que novos estudos são necessários para elucidar se a ART e CSP podem ocasionar a MCI ou apoptose em linhagens celulares de LO e CCEO (AU)

New drugs, such as artemisinin (ART), may be promising in the treatment of potentially malignant lesions (PML) and may be useful when used in combination with other chemotherapy drugs, especially in reducing their side effects. Oral leukoplakia (OL) is the most common LPM of the oral cavity and can progress to oral squamous cell carcinoma (OSCC). There is no therapy to prevent its malignant transformation, chemoprevention can initiate immunogenic cell death (ICM) that activates the immune system to recognize and eliminate malignant or pre-malignant cells, being a potential treatment for LPM. The present study aimed to evaluate whether or not ART and cisplatin (CSP) combined would be capable of inducing MCI in LO (DOK) and CCEO SCC-180) cell lines. Material and methods: HaCat (control), DOK and SCC-180 cell lines were treated by ART and CSP in combination or alone, in order to analyze the cytotoxicity and genotoxicity of these drugs, in addition to their ability to reduce cell and, whether the compounds would be able to induce the expression of high mobility box protein (HGMB-1), caspase 3, 8, 9, and Calreticulin (CALR). Results: In cytotoxicity at higher doses, which was not observed with ART. The formation of micronuclei was not observed in the genotoxicity test. The migration rate was reduced with the IC50 concentrations of CSP and ART+CSP for OSCC cells. No significant expressions of proteins related to MCI or apoptosis were found in the LO and CCEO lines, treated with ART, CSP or ART+CSP, indicating that another type of cell death may have occurred. Conclusion: MCI and apoptosis were not evidenced as a form of cell death after the LO and CCEO lines received treatments with ART, CSP and the combination of both. Genotoxic effects were not observed at the doses tested. CSP and ART+CSP treatment was able to reduce OSCC cell migration. We also conclude that new studies are necessary to elucidate whether ART and CSP can cause MCI or apoptosis in LO and CCEO cell lines.(AU)

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The formation of oral structures occurs due to ectomesenchymal cell proliferation, which will originate an anatomical structure known as peri coronal follicle, which undergoes apoptosis after complete formation of the dental crown, causing its eruption. In some cases, there may be impaction of the tooth as a result of an alteration in the reduced epithelium of the enamel organ, which prevents epithelial self-destruction. These changes may be associated with blood type which, based on scientific findings, can prove a correlation between human diseases and the ABO blood system. Thus, the present research project aims to find out if there is a relationship between the anatomical structure of the peri coronal follicle and the dentigerous cyst, comparing it with the blood typing of the individuals who carry it. The research is descriptive with data collection of cases of peri coronal follicle and dentigerous cyst, sent to the histopathological laboratory of the University of Passo Fundo. The prevalence of the ABO blood system found for the anatomical alteration of the peri coronal follicle was A+ and for the pathological structure of the dentigerous cyst it was O+. The other classes, AB, were presented in a smaller percentage, with B+, B-, A- only in the peri coronal follicle, as well as O-. These results conclude that blood typing can establish a predisposition for odontogenic findings, requiring further research to confirm the clinical diagnosis. (AU)

A formação das estruturas bucais ocorre devido às proliferações celulares ectomesenquimais, o que irão originar uma estrutura anatômica conhecida como folículo pericoronário, a qual sofre apoptose após a formação completa da coroa dentária, ocasionando sua erupção. Em alguns casos, pode haver a impactação do dente por consequência de uma alteração no epitélio reduzido do órgão do esmalte, o que impede a autodestruição epitelial. Essas alterações podem estar associadas ao tipo sanguíneo que, com base em achados científicos, pode-se provar uma correlação entre doenças humanas e o sistema sanguíneo ABO. Assim, o presente projeto de pesquisa tem por objetivo averiguar se há relação entre a estrutura anatômico do folículo pericoronário e o cisto dentígero comparando com a tipagem sanguínea dos indivíduos que o portam. A pesquisa é descritiva com coleta de dados dos casos de folículo pericoronário e cisto dentígero, enviados para o laboratório histopatológico da Universidade de Passo Fundo. A prevalência do sistema sanguíneo ABO encontrada para a alteração anatômico do folículo pericoronário foi A+ e para estrutura patológica do cisto dentígero foi O+. As demais classes, AB, apresentaram-se em menor porcentagem, sendo B+, B-, A- apenas em folículo pericoronário, assim como o-. Tais resultados concluem que a tipagem sanguínea pode estabelecer uma pré-disposição para os achados odontogênicos, sendo necessário maiores pesquisas para confirmação de diagnóstico clínico. (AU)

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[This corrects the article DOI: 10.3389/fimmu.2023.1241038.].

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The SARS CoV-2 antibody and CD4+ T cell responses induced by natural infection and/or vaccination decline over time and cross-recognize other viral variants at different levels. However, there are few studies evaluating the levels and durability of the SARS CoV-2-specific antibody and CD4+ T cell response against the Mu, Gamma, and Delta variants. Here, we examined, in two ambispective cohorts of naturally-infected and/or vaccinated individuals, the titers of anti-RBD antibodies and the frequency of SARS-CoV-2-specific CD4+ T cells up to 6 months after the last antigen exposure. In naturally-infected individuals, the SARS-CoV-2 antibody response declined 6 months post-symptoms onset. However, the kinetic observed depended on the severity of the disease, since individuals who developed severe COVID-19 maintained the binding antibody titers. Also, there was detectable binding antibody cross-recognition for the Gamma, Mu, and Delta variants, but antibodies poorly neutralized Mu. COVID-19 vaccines induced an increase in antibody titers 15-30 days after receiving the second dose, but these levels decreased at 6 months. However, as expected, a third dose of the vaccine caused a rise in antibody titers. The dynamics of the antibody response upon vaccination depended on the previous SARS-CoV-2 exposure. Lower levels of vaccine-induced antibodies were associated with the development of breakthrough infections. Vaccination resulted in central memory spike-specific CD4+ T cell responses that cross-recognized peptides from the Gamma and Mu variants, and their duration also depended on previous SARS-CoV-2 exposure. In addition, we found cross-reactive CD4+ T cell responses in unexposed and unvaccinated individuals. These results have important implications for vaccine design for new SARS-CoV-2 variants of interest and concern.

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OBJECTIVE: To evaluate the impact of body mass index associated with advanced maternal age on pregnancy outcomes. METHODS: A retrospective and observational study that included 808 in vitro fertilization cycles and evaluated: age, weight, height, number of oocytes and mature oocytes, number of embryos and transferred embryos, fertilization and clinical pregnancy rates. Four categories of body mass index: underweight, adequate weight, overweight and obesity. We classified age into 4 categories: 35-37; 38-40; 41-42 and over 42 years of age. The means and rates were calculated and compared between different ages and body mass index groups. RESULTS: For the fresh group, women who achieved clinical pregnancy had a lower mean age than those who did not become pregnant, being the higher the pregnancy rate the lower the age (p<0.0001). After logistic regression analysis for data associated with clinical pregnancy in the fresh group, the number of transferred embryos remained higher in the overweight category (p=0.0001). Overweight and obese women had a significantly higher rate of mature oocytes when compared with adequate weight (p=0.015). Analysis using the ROC curve indicated an area under the curve of 60% (p=0.002) for the fresh group. CONCLUSIONS: The adverse effect of high BMI on clinical pregnancy rates is greater in women under 35 years compared to older women; and age had a higher impact on live birth rate rather than BMI, when the analysis was performed on older women, with the impact of BMI on the probability of having a live birth depending on maternal age.

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Although well-established and adopted by commercial laboratories, the in vitro embryo production system still requires refinements to achieve its highest efficiency. Early embryonic development is a dynamic event, demanding suitable conditions to provide a high number of embryos with quality and competence. The first step to obtaining an optimized in vitro environment is to know the embryonic metabolism and energy request throughout the different stages of development. Oxygen plays a crucial role in several key biological processes necessary to sustain and complete embryonic development. Nonetheless, there is still controversy regarding the optimal in vitro atmospheric concentrations during culture. Herein, we discuss the impact of oxygen tension on the viability of in vitro-produced embryos during early development. The importance of oxygen tension is addressed as its roles regarding essential embryonic traits, including embryo production rates, embryonic cell viability, gene expression profile, epigenetic regulation, and post-cryopreservation survival. Finally, we highlight the damage caused by in vitro unbalanced oxygen tensions and strategies to mitigate the harmful effects.

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Two studies evaluated the productive characteristics of young Nellore heifers receiving different days on feed (DOF) to determine the biological slaughter endpoint. In Experiment 1 (Exp. 1), fifty-one Nellore heifers [324 ± 19.3 kg of body weight (BW); 16 ± 1 months] were split into three DOF lengths (45, 75, or 105 days), while in Experiment 2 (Exp. 2), thirty-six Nellore heifers (362 ± 25.5 kg of BW; 18 ± 1 months of age) were split into three DOF lengths (45, 90, or 135 days). In both studies, all animals were distributed in complete randomized blocks according to initial BW and stratified via carcass ultrasound. The diet was supplied ad libitum, allowing 3% of refusals. The point at which the animals achieved 25% of ether extract of shrunk body weight (EESBW) was defined as the biological endpoint. Thus, relationships were made between some characteristics obtained in both studies. Positive linear relationships were found between backfat thickness (BFT) vs. EESBW (p < 0.001, r = 0.84) and BFT vs. body weight (p < 0.001, r = 0.77). Our results suggest that the biological slaughter endpoint for young Nellore heifers is 6.97 mm of backfat thickness or 402 kg shrunk body weight, corresponding to around 75 DOF.

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Each living organism is unique because of the lipid identity of its organelles. The diverse distribution of these molecules also contributes to the role of each organelle in cellular activity. The lipid profiles of whole embryos are well documented in the literature. However, this approach can often lead to the loss of relevant information at the subcellular and consequently, metabolic levels, hindering a deeper understanding of key physiological processes during preimplantation development. Therefore, we aimed to characterize four organelles in vitro-produced bovine embryos: lipid droplets (LD), endoplasmic reticulum (ER), mitochondria (MIT), and nuclear membrane (NUC), and evaluate the contribution of the lipid species to each organelle evaluated. Expanded blastocysts were subjected to cell organelle isolation. Thereafter, lipid extraction from cell organelles and lipid analysis using the Multiple Reaction Monitoring (MRM) profiling method were performed. The LD and ER displayed a greater number of lipids (Phosphatidylcholine - PC, Ceramide - Cer, and Sphingomielin - SM) with high signal-to-noise intensities. This result is due to the high rate of biosynthesis, lipid distribution, and ability to store and recycle lipid species of these organelles. The NUC had a more distinct lipid profile than the other three organelles, with high relative intensities of PC, SM, and triacylglycerols (TG), which is consistent with its high nuclear activity. MIT had an intermediate profile that was close to that of LD and ER, which aligns with its autonomous metabolism for some classes of phospholipids (PL). Our study revealed the lipid composition of each organelle studied, and the roles of these lipids could be associated with the characteristic organellar activity. Our findings highlight the lipid species and classes that are relevant for the homeostasis and function of each associated organelle and provide tentative biomarkers for the determination of in vitro embryonic development and quality.

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Reactive dairy cows are more susceptible to stress, and this may result in negative effects on milk yield and quality. The aims of this study were to investigate the relationships between temperament traits and concentration of milk cortisol and oxytocin, milk yield, milkability, and milk quality in Holstein-Gyr cows. Temperament traits were assessed in 76 Holstein-Gyr cows in the milking parlor (by scoring milking reactivity and recording the numbers of steps and kicks during pre-milking udder preparation and when fitting the milking cluster) and during handling in the corral (by measuring the time to enter in the squeeze chute, ET and flight speed, FS). Milk samples were collected for milk quality (% fat, % protein, % lactose, and somatic cell count, SCC), and milk cortisol and oxytocin. Milk yield, milking time, and average flow were also measured. The calmer cows during milking management (class 'low') produced milk with higher protein (p = 0.028) content and tendencies for lower fat (p = 0.056) and higher lactose (p = 0.055) contents. Regarding the hormones, the most reactive cows (class 'high') in the milking and handling corral produced milk with higher concentrations of cortisol (p<0.001) and oxytocin (p = 0.023). In addition, the temperament of the animals affected some of the productive measures evaluated. Cows with reactive temperament had lower milk flow and longer milking time than the intermediate ones and had higher fat and a tendency for lower protein percentage in milk compared to cows with intermediate temperaments. Calm and intermediate cows in the handling corral produced more milk and presented better milkability parameters, such as a shorter milking time and greater average milk flow. Our results suggest that the cows' behavioral reactivity can be related to the intensity of their response to stress during handling.

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BACKGROUND: The use of non-invasive positive pressure ventilation (NIPPV) in COVID-19 patients with hypoxaemia is still under debate. The aim was to evaluate the efficacy of NIPPV (CPAP, HELMET-CPAP or NIV) in COVID-19 patients treated in the dedicated COVID-19 Intermediate Care Unit of Coimbra Hospital and University Centre, Portugal, and to assess factors associated with NIPPV failure. METHODS: Patients admitted from December 1st 2020 to February 28th 2021, treated with NIPPV due to COVID-19 were included. Failure was defined as orotracheal intubation (OTI) or death during hospital stay. Factors associated with NIPPV failure were included in a univariate binary logistic regression analysis; those with a significance level of p < 0.001 entered a multivariate logistic regression model. RESULTS: A total of 163 patients were included, 64.4% were males (n = 105). The median age was 66 years (IQR 56-75). NIPPV failure was observed in 66 (40.5%) patients, 26 (39.4%) were intubated and 40 (60.6%) died during their hospital stay. The highest CRP (OR 1.164; 95%CI 1.036-1.308) and morphine use (OR 24.771; 95%CI 1.809-339.241) were identified as predictors of failure after applying multivariate logistic regression. Adherence to prone positioning (OR 0.109; 95%CI 0.017-0.700) and a higher value of the lowest platelet count during hospital stay (OR 0.977; 95%CI 0.960-0.994) were associated with a favorable outcome. CONCLUSIONS: NIPPV was successful in over half of patients. Highest CRP during hospital stay and morphine use were predictors of failure. Adherence to prone positioning and a higher value of the lowest platelet count during hospital stay were associated with a favourable outcome.

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The objective of this study is to understand how the type of wound dressing changes (routine or frequent) in patients admitted to intensive care units influences nurses' workload. This study used a database of retrospective and analytical observational study from one Portuguese intensive care unit. The sample included 728 adult patients admitted between 2015 and 2019. The nursing workload was assessed by the TISS-28 scale, both at admission and at discharge. The linear regression results show that patients with frequent dressing changes are associated with a higher nursing workload, both at admission (Coef. 1.65; 95% CI [0.53; 2.77]) and discharge (Coef. 1.27; 95% CI [0.32; 2.22]). In addition, age influences the nursing workload; older people are associated with a higher nursing workload (at admission Coef. 0.07; 95% CI [0.04; 0.10]; at discharge Coef. 0.08; 95% CI [0.05; 0.10]). Additionally, an increase in nursing workload at admission would significantly increase the nursing workload at discharge (Coef. 0.27; 95% CI [0.21; 0.33]). The relative stability of the nursing workload over the studied years is also another important finding (the influence of studied years is non-significant). In conclusion, patients with frequent dressing changes presented higher TISS-28 scores when compared with patients with an exchange of routine dressings, which leads to a higher nursing workload.

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A wide-ranging review study regarding the molecular characterization of the first cell lineages of the developmental embryo is lacking, especially for the primary events during earliest differentiation which leads to the determination of cellular fate. Here, a systematic review and meta-analysis were conducted according to PRISMA guidelines. MEDLINE-PubMed was searched based on an established search strategy through April 2021. Thirty-six studies fulfilling the inclusion criteria were subjected to qualitative and quantitative analysis. Among the studies, 50 % (18/36) used mice as an animal model, 22.2 % (8/36) pigs, 16.7 % (6/36) cattle, 5.5 % (2/36) humans, and 2.8 % (1/36) goats as well as 2.8 % (1/36) equine. Our results demonstrated that each of the first cell lineages of embryos requires a certain pattern of expression to establish the cellular determination of fate. Moreover, these patterns are shared by many species, particularly for those molecules that have already been identified in the literature as biomarkers. In conclusion, the present study integrated carefully chosen studies regarding embryonic development and first cellular decisions in mammalian species and summarized the information about the differential characterization of the first cell lineages and their possible relationship with specific gene expression.

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