Clinically Expired Platelet Concentrates as a Source of Extracellular Vesicles for Targeted Anti-Cancer Drug Delivery.

The short shelf life of platelet concentrates (PC) of up to 5-7 days leads to higher wastage due to expiry. To address this massive financial burden on the healthcare system, alternative applications for expired PC have emerged in recent years. Engineered nanocarriers functionalized with platelet membranes have shown excellent targeting abilities for tumor cells owing to their platelet membrane proteins. Nevertheless, synthetic drug delivery strategies have significant drawbacks that platelet-derived extracellular vesicles (pEV) can overcome. We investigated, for the first time, the use of pEV as a carrier of the anti-breast cancer drug paclitaxel, considering it as an appealing alternative to improve the therapeutic potential of expired PC. The pEV released during PC storage showed a typical EV size distribution profile (100-300 nm) with a cup-shaped morphology. Paclitaxel-loaded pEV showed significant anti-cancer effects in vitro, as demonstrated by their anti-migratory (>30%), anti-angiogenic (>30%), and anti-invasive (>70%) properties in distinct cells found in the breast tumor microenvironment. We provide evidence for a novel application for expired PC by suggesting that the field of tumor treatment research may be broadened by the use of natural carriers.

Single-walled silicon nanotube as an exceptional candidate to eliminate SARS-CoV-2: a theoretical study.

In this work, computational chemistry methods were used to study a silicon nanotube (Si192H16) as possible virucidal activity against SARS-CoV-2. This virus is responsible for the COVID-19 disease. DFT calculations showed that the structural parameters of the Si192H16 nanotube are in agreement with the theoretical/experimental parameters reported in the literature. The low energy gap value (0.29 eV) shows that this nanotube is a semiconductor and exhibits high reactivity. For nanomaterials to be used as virucides, they need to have high reactivity and high inhibition constant values. Therefore, the adsorption of 3O2 and H2O on the surface of Si192H16 (Si192H16@O2-H2O) was performed. In this process, the formation and activation energies were -51.63 and 16.62 kcal/mol, respectively. Molecular docking calculations showed that the Si192H16 and Si192H16@O2H-OH nanotubes bind favorably on the receptor-binding domain of the SARS-CoV-2 spike protein with binding energy of -11.83 (Ki = 2.13 nM) and -11.13 (Ki = 6.99 nM) kcal/mol, respectively. Overall, the results obtained herein indicate that the Si192H16 nanotube is a potential candidate to be used against COVID-19 from reactivity process and/or steric impediment in the S-protein.Communicated by Ramaswamy H. Sarma.

Work hours as a risk factor for SARS-CoV-2 infections: cardiometabolic and sleep characteristics in rotating shift workers.

Objective: This study aims to describe the health characteristics of rotating shift mining workers that may be related to a worse course scenario for COVID-19, according to literature data. Methods: Is a cross-sectional from three studies with 1478 shift workers. Social, demographic, clinical, and biochemical variables were analyzed. Risk factors for COVID-19 analyzed: hyperglycemia, altered blood pressure, dyslipidemia, hypovitaminosis D, obesity, presence of pre-existing cardiovascular diseases, and smokers. Results: Evaluating the grouped risk factors for an unfavorable evolution of COVID-19 most workers (91.0%) presented at least one risk factor. Discussion: With coronavirus in pandemic circulation, we noticed that mineworkers are in a vulnerable position. Their exposure to occupational risk factors, to the shift system, which directly affects sleep and negatively influences immunity, added to the conditions of favorable transmissibility by the flow of people from the mines leads us to believe in their greater susceptibility to acquiring the most serious forms of the disease.

Biological Therapy in Patients with Rheumatoid Arthritis in a Tertiary Center in Portugal: A Cross-Sectional Study.

INTRODUCTION: Clinical outcomes in rheumatoid arthritis have greatly improved with therapeutic advances. Despite the availability of substantial clinical trial evidence, there is a lack of real-life data. The aim of this study was to assess disease status and quality of life in an outpatient population treated with biological disease-modifying anti-rheumatic drugs. MATERIAL AND METHODS: Cross-sectional study recalling all patients ever treated in our unit with biological disease-modifying antirheumatic drugs. Clinical and demographic data, compliance, disease activity, functional status, joint deformities, and comorbidities were documented, and patients queried on occupational status, education, marital status and generic health related quality of life questionnaires. RESULTS: Recall was attended by 77 of the original 94 patients. At recall, median age was 63 years old, 82% of the patients were female and the median disease duration was 12 years. Biological therapy was started at a median of four years following disease onset. According to the disease activity score (DAS28), the percentage of patients with high, moderate, low disease activity or remission changed from 50, 45, 0 and 5 (pre-therapy) to 11, 37, 25 and 26 at recall, respectively; functional status was significantly improved. Seventy-five per cent of the patients retained the original treatment with good compliance. Lower Short Form-36 domain scores accompanied a low EQ-5D-3L score. Deceased patients (n = 6) had a lower estimated 10-year survival rate. In this group, biological therapy was discontinued at a higher frequency during follow-up. DISCUSSION: A high disease activity and a high HAQ disability index characterized most patients at pre-bDMARD onset. CONCLUSION: Despite therapy switches and regular follow-up, a significant percentage of patients still presented with moderate disease activity, functional impairment and a poor health-related quality of life.

Introdução: Avanços no tratamento da artrite reumatóide contribuiram para uma evolução favorável. Apesar de evidências substanciais provenientes de ensaios clínicos, são menos conhecidos os dados provenientes da vida real. O objetivo do estudo foi caracterizar a doença e a qualidade de vida em doentes sob fármacos biotecnológicos. Material e Métodos: Trata-se de um estudo transversal com recolha de dados clínicos relativos à adesão terapêutica, atividade da doença, capacidade funcional, deformidades articulares, comorbilidades e questionários de qualidade de vida relacionada com a saúde, estado civil, situação profissional e escolaridade. Resultados: Foram recrutados 77 doentes do grupo original de um total de 94. A mediana da idade foi 63 anos, 82% do sexo feminino e início de biológico cerca de quatro anos após o início da doença, com uma mediana de duração de 12 anos. De acordo com o disease activity score (DAS28), a percentagem de doentes com atividade alta, moderada, baixa ou em remissão mudou, respectivamente, de 50, 45, 0 e 5 pré- biológico para 11, 37, 25 e 26 na altura da re-avaliação, com melhoria funcional. Setenta e cinco por cento dos doentes mantiveram o tratamento original com boa adesão. Pontuações mais baixas do short form-36 associaram-se a uma baixa pontuação no EQ-5D-3L. No grupo de doentes que viriam a falecer (n = 6), foi observada uma menor esperança de vida aos 10 anos, assim como uma maior discontinuação da terapêutica biológica. Discussão: Pré-biológico, uma elevada percentagem dos doentes apresentava elevada atividade da doença e incapacidade funcional. Conclusão: Não obstante ajustes terapêuticos e seguimento regular, uma percentagem significativa de doentes mantinha atividade moderada e limitação funcional com baixa qualidade de vida relacionada com a saúde.

Anticancer Activities of the Quinone-Methide Triterpenes Maytenin and 22-ß-hydroxymaytenin Obtained from Cultivated Maytenus ilicifolia Roots Associated with Down-Regulation of miRNA-27a and miR-20a/miR-17-5p.

Natural triterpenes exhibit a wide range of biological activities. Since this group of secondary metabolites is structurally diverse, effects may vary due to distinct biochemical interactions within biological systems. In this work, we investigated the anticancer-related activities of the quinone-methide triterpene maytenin and its derivative compound 22-ß-hydroxymaytenin, obtained from Maytenus ilicifolia roots cultivated in vitro. Their antiproliferative and pro-apoptotic activities were evaluated in monolayer and three-dimensional cultures of immortalized cell lines. Additionally, we investigated the toxicity of maytenin in SCID mice harboring tumors derived from a squamous cell carcinoma cell line. Both isolated molecules presented pronounced pro-apoptotic activities in four cell lines derived from head and neck squamous cell carcinomas, including a metastasis-derived cell line. The molecules also induced reactive oxygen species (ROS) and down-regulated microRNA-27a and microRNA-20a/miR-17-5p, corroborating with the literature data for triterpenoids. Intraperitoneal administration of maytenin to tumor-bearing mice did not lead to pronounced histopathological changes in kidney tissue, suggesting low nephrotoxicity. The wide-ranging activity of maytenin and 22-ß-hydroxymaytenin in head and neck cancer cells indicates that these molecules should be further explored in plant biochemistry and biotechnology for therapeutic applications.

Mandibular resorption and vocal cord paralysis: a catastrophic form of systemic sclerosis.

Sudden respiratory distress in association with severe weight loss are unusual features of systemic sclerosis (SSc). We report the case of a 56-year-old Caucasian woman with a 9-year history of a diffuse form of SSc who presented with acute stridor due to vocal cord paralysis and required an emergency tracheostomy. She had sought medical attention only after 4 years of disease onset, presenting with a mask-like face, diffuse skin thickening, acro-osteolysis and severe interstitial lung disease. Even though skin tightness improved after immunosuppressive treatment, several spontaneous facial fractures and episodes of dysphagia and choking occurred in the years that followed. At the time of stridor, she was severely malnourished and a percutaneous endoscopic gastrostomy was required for feeding. Permanent vocal cord damage in combination with severe loco-regional bone resorption resulted in severe disability and impaired nutrition. We hereby highlight the features of SSc for which therapy remains challenging.

The tyrosine kinase inhibitor nilotinib is more efficient than mitotane in decreasing cell viability in spheroids prepared from adrenocortical carcinoma cells.

BACKGROUND: New drugs for adrenocortical carcinoma (ACC) are needed because most patients undergo rapid disease progression despite surgery and adjuvant therapy with mitotane. In this study, we aimed to investigate the in vitro effects of different chemotherapy drugs, alone or combined with mitotane, on the viability of adrenocortical carcinoma cells. METHODS: Everolimus, sunitinib, zoledronic acid, imatinib and nilotinib cytotoxicity, alone or combined with mitotane were tested on ACC H295R cells in monolayer or spheroid cultures using MTS assays and confocal microscopy. Moreover, the nilotinib effects were investigated in spheroids cultured from patient tumor-derived ACC-T36 cells. RESULTS: Morphological characterization of H295R cell spheroids using histochemistry was performed and showed that dense, homogenously sized, multicellular spheroids were obtained. We observed that sunitinib and nilotinib alone were equally effective in a monolayer preparation, whereas mitotane was the most effective even at a low dose. A combination of sunitinib and mitotane was the most effective treatment, with only 23.8% of cells in the monolayer remaining viable. Spheroid preparations showed resistance to different drugs, although the poor effect produced by mitotane alone was surprising, with a cell viability of 84.6% in comparison with 13.1% in monolayer cells. The most ineffective drugs in spheroid preparations were everolimus, zoledronic acid and imatinib. In both cell types, nilotinib, either alone or in combination with mitotane induced more significant cell viability inhibition in monolayer and spheroid preparations. In addition, the mechanism of nilotinib activity involves the ERK1/2 pathway. CONCLUSION: Taken together, our data identified nilotinib as a cytotoxic drug that combined with ERK inhibitors deserves to be tested as a novel therapy for adrenocortical carcinoma.

SOCS3 expression in SF1 cells regulates adrenal differentiation and exercise performance.

Many hormones/cytokines are secreted in response to exercise and cytokine signaling may play a pivotal role in the training adaptations. To investigate the importance of cytokine signaling during vertical ladder climbing, a resistance exercise model, we produced mice lacking SOCS3 protein exclusively in steroidogenic factor-1 (SF1) cells (SF1 Socs3 KO mice). SF1 expression is found in steroidogenic cells of the adrenal cortex and gonads, as well as in neurons of the ventromedial nucleus of the hypothalamus. Histological markers of the fetal adrenal zone (or X-zone in rodents) were still present in adult males and postpartum SF1 Socs3 KO females, suggesting a previously unrecognized effect of SOCS3 on the terminal differentiation of the adrenal gland. This change led to a distinct distribution of lipid droplets along the adrenal cortex. Under basal conditions, adult SF1 Socs3 KO mice exhibited similar adrenal weight, and plasma ACTH and corticosterone concentrations. Nonetheless, SF1 Socs3 KO mice exhibited a blunted ACTH-induced corticosterone secretion. The overall metabolic responses induced by resistance training remained unaffected in SF1 Socs3 KO mice, including changes in body adiposity, glucose tolerance and energy expenditure. However, training performance and glucose control during intense resistance exercise were impaired in SF1 Socs3 KO mice. Furthermore, a reduced counter-regulatory response to 2-deoxy-d-glucose was observed in mutant mice. These findings revealed a novel participation of SOCS3 regulating several endocrine and metabolic aspects. Therefore, cytokine signaling in SF1 cells exerts an important role to sustain training performance possibly by promoting the necessary metabolic adjustments during exercise.

Canine Macrophage DH82 Cell Line As a Model to Study Susceptibility to Trypanosoma cruzi Infection.

Trypanosoma cruzi is an obligatory intracellular protozoan parasite, and it is the etiological agent of Chagas' disease that is endemic in the Americas. In addition to humans, a wide spectrum of mammals can be infected by T. cruzi, including dogs. Dogs develop acute and chronic disease, similar to human infection. T. cruzi can infect almost all cell types and after cell invasion, the metacyclics trypomastigotes localize in the cytoplasm, where they transform into amastigotes, the replicative form of T. cruzi in mammals. After amastigote multiplication and differentiation, parasites lyse host cells and spread through the body by blood circulation. In this work, we evaluated the in vitro ability of T. cruzi to infect a canine macrophage cell line DH82 compared with RAW264.7, a murine tissue culture macrophage. Our results have shown that the T. cruzi is able to infect, replicate and differentiate in DH82 cell line. We observed that following treatment with LPS and IFN-γ DH82 cells were more resistant to infection and that resistance was not related reactive oxygen species production in our system. In this study, we also found that DH82 cells became more susceptible to T. cruzi infection when cocultured with apoptotic cells. The analysis of cytokine production has showed elevated levels of the TGF-ß, IL-10, and TNF-α produced by T. cruzi-infected canine macrophages. Additionally, we demonstrated a reduced expression of the MHC class II and CD80 by infected DH82 cell line.

Insights from Systems Biology in Physiological Studies: Learning from Context.

Systems biology presents an integrated view of biological systems, focusing on the relations between elements, whether functional or evolutionary, and providing a rich framework for the comprehension of life. At the same time, many low-throughput experimental studies are performed without influence from this integrated view, whilst high-throughput experiments use low-throughput results in their validation and interpretation. We propose an inversion in this logic, and ask which benefits could be obtained from a holistic view coming from high-throughput studies-and systems biology in particular-in interpreting and designing low-throughput experiments. By exploring some key examples from the renal and adrenal physiology, we try to show that network and modularity theory, along with observed patterns of association between elements in a biological system, can have profound effects on our ability to draw meaningful conclusions from experiments.

Comparative Effect of ACTH and Related Peptides on Proliferation and Growth of Rat Adrenal Gland.

Pro-opiomelanocortin (POMC) is a polypeptide precursor known to yield biologically active peptides related to a range of functions. These active peptides include the adrenocorticotropic hormone (ACTH), which is essential for maintenance of adrenal growth and steroidogenesis, and the alpha-melanocyte stimulation hormone, which plays a key role in energy homeostasis. However, the role of the highly conserved N-terminal region of POMC peptide fragments has begun to be unraveled only recently. Here, we review the cascade of events involved in regulation of proliferation and growth of murine adrenal cortex triggered by ACTH and other POMC-derived peptides. Key findings regarding signaling pathways and modulation of genes and proteins required for the regulation of adrenal growth are summarized. We have outlined the known mechanisms as well as future challenges for research on the regulation of adrenal proliferation and growth triggered by these peptides.

Diferentes estratégias alimentares na larvicultura do acará-bandeira (Peterolophyllum scalare,Cichlidae) / Different feeding strategies on larval rearing of angelfish (Peterolophyllum scalare, Cichlidae)

The hatchery is one of the most critical stages of the development of fish and their success is directly related to food handling that can provide greater survival and growth. The aim of this study was to evaluate the time of dietary transition and providing meta-nauplii of Artemia spp. (MNA) for the angelfish larvae. Two experiments were conducted in a completely randomized design with five treatments and four repetitions. On each experiment there were used 540 fishes, distributed in 20 tanks with 2 liters each. In the first experiment, there were evaluated the dietary transition periods (MNA + Diet) for 1, 2, 3, 4 and 5 days. In the second experiment, there were evaluated the supply of MNA for 5, 10, 15, 20 and 25 days. There were evaluated: weight gain, growth rate and specific development, survival and batch uniformity (only in the experiment to evaluate the time of fish feed with MNA). There was no significant effect of different periods of dietary transition on the growth variables (p>0.05), but survival was higher (p<0.05) in the treatments composed of 3, 4 and 5 days of joint feed. In relation to the time of supply of MNA worse results were observed (p<0.05) when the time of supply of live food was shorter (5, 10 and 15 days). The animals were fed with MNA before feeding transition, for longer time (20 and 25) showed the best results for growth (p<0.05). Therefore, a food transition of three days and a supply of MNA for 20 days is recommended to perform the total replacement of live food for feed.

A larvicultura é uma das etapas mais críticas do desenvolvimento dos peixes e o seu sucesso está diretamente relacionado ao manejo alimentar, que pode proporcionar maiores sobrevivência e crescimento. Objetivou-se avaliar o tempo de transição alimentar e de fornecimento de meta-náuplios de Artemia spp (MNA) na larvicultura do acará-bandeira. Dois experimentos foram conduzidos em delineamento inteiramente casualizado com cinco tratamentos e quatro repetições. Em cada experimento foram utilizados 540 peixes distribuídos em 20 aquários com 2 L. No primeiro experimento, avaliaram-se os períodos de transição alimentar (MNA + ração) por 1, 2, 3, 4 e 5 dias. No segundo experimento, avaliou-se o período de fornecimento de MNA por 5, 10, 15, 20 e 25 dias. Foram avaliados: ganho de peso, taxas de crescimento e desenvolvimento específico, sobrevivência e uniformidade do lote (apenas no experimento para avaliar o tempo de fornecimento de MNA). Não houve efeito significativo dos diferentes períodos de transição alimentar sobre as variáveis de crescimento (p>0,05), porém a sobrevivência foi maior (p<0,05) nos tratamentos compostos por 3, 4 e 5 dias de alimentação conjunta. Em relação ao tempo de fornecimento de MNA, foram observados piores resultados (p<0,05) quando o tempo de fornecimento do alimento vivo foi menor (5, 10 e 15 dias). Os animais que foram alimentados com MNA antes da transição alimentar, por mais tempo (20 e 25 dias), apresentaram os melhores resultados de crescimento (p<0,05). Portanto, recomenda-se uma transição alimentar de três dias e um fornecimento de MNA por 20 dias para realizar a substituição total do alimento vivo pela ração.

The involvement of Nek2 and Notch in the proliferation of rat adrenal cortex triggered by POMC-derived peptides.

The adrenal gland is a dynamic organ that undergoes constant cell turnover. This allows for rapid organ remodeling in response to the physiological demands of the HPA axis, which is controlled by proopiomelanocortin (POMC)-derived peptides, such as adrenocorticotropic hormone (ACTH) and N-Terminal peptides (N-POMC). In the rat adrenal cortex, POMC-derived peptides trigger a mitogenic effect, and this process increases cyclins D and E, while inhibiting p27Kip1. The goal of the present study was to further explore the mitogenic effect of ACTH and synthetic N-POMC1-28 peptides by investigating the differences in the expression of key genes involved in the cell cycle of the rat adrenal cortex, following inhibition of the HPA axis. Moreover, we evaluated the differences between the inner and outer fractions of the adrenal cortex (ZF-fraction and ZG-fraction) in terms of their response patterns to different stimuli. In the current study, the inhibition of the HPA axis repressed the expression of Ccnb2, Camk2a, and Nek2 genes throughout the adrenal cortex, while treatments with POMC-derived peptides stimulated Nek2, gene and protein expression, and Notch2 gene expression. Furthermore, Notch1 protein expression was restricted to the subcapsular region of the cortex, an area of the adrenal cortex that is well-known for proliferation. We also showed that different regions of the adrenal cortex respond to HPA-axis inhibition and to induction with POMC-derived peptides at different times. These results suggest that cells in the ZG and ZF fractions could be at different phases of the cell cycle. Our results contribute to the understanding of the mechanisms involved in cell cycle regulation in adrenocortical cells triggered by N-POMC peptides and ACTH, and highlight the involvement of genes such as Nek2 and Notch.

A suplementação de vitamina C na estruturação do tecido conjuntivo de melanotência-maçã / Vitamin C supplementation in the structuring of connective tissue of red rainbowfish

Peixes cultivados mostram-se altamente sensíveis a dietas deficientes em ácido ascórbico com sinais clínicos que afetam o desempenho e a comercialização por provocar deformidades. Com objetivo de conhecer os níveis mínimos para a melanotênia-maçã, foram testadas seis rações isoaminoacídicas e isocalóricas com níveis variados de vitamina C ativa. Ao final dos 42 dias experimentais, 36 peixes foram analisados histologicamente quanto à porcentagem de tecido conjuntivo e fibras musculares a fim de determinar a influência da porcentagem de Vitamina C nesses índices. Apesar de as médias serem estatisticamente iguais do ponto de vista quantitativo, qualitativamente os níveis mais altos de vitamina C possibilitaram melhor estrutura tecidual. A desestruturação do tecido muscular é um indicativo da deficiência alimentar com ácido ascórbico...

Farmed fishes are highly sensitive to diets deficient in ascorbic acid with clinical symptoms that affect performance and commercialization for causing deformities. Aiming to meet the minimum levels for red rainbow fish, six isoaminoacid and isocaloric diets with varying levels of active vitamin C were tested. At the end of 42 days experimental period, 36 fishes were histologically analyzed for the percentage of connective tissue and muscle fibers in order to determine the influence of the Vitamin C percentage in those ratios. Although the averages are statistically indistinguishable from the quantitative point of view, qualitatively higher levels of vitamin C allow better tissue structure. The breakdown of muscle tissue is indicative for dietary deficiency of ascorbic acid...

Health professionals moving to... and from Portugal.

The mobility of health professionals in the European Union is a phenomenon which policy-makers must take into account to provide the conditions to adjust for demand and supply of health services. This paper presents the case of Portugal, a country which at the same time imports and exports health workers. Since the early 1990s Portugal became a destination country receiving foreign health care professionals. This situation is now changing with the current economic situation as fewer immigrants come and more Portuguese emigrate. Foreigners coming to Portugal do so in part for similar reasons that bring Portuguese to want to emigrate, mainly the search for better work conditions and professional development opportunities. The emigration of Portuguese health professionals is also stimulated by the difficulty for recently graduated nurses, dentists and diagnostic and therapeutic technicians to find employment, low salaries in the public and private sectors, heavy workloads, remuneration not related to performance and poor career prospects. The paradoxes described in this study illustrate the consequences of the absence of a policy for the health professions. Strategies based on evidence, and on an integrated information system that captures the dynamic evolution of the workforce in health are not only necessary but also a good investment.

N-POMC1-28 increases cyclin D expression and inhibits P27(kip1) in the adrenal cortex.

The Adrenocorticotropic hormone (ACTH) and Pro-opimelanocortin (POMC) 1-28N-terminal peptide (N-POMC(1-28)) have been shown to act as an adrenal mitogen in vivo. A possible role for cyclin E in the zona glomerulosa (ZG) proliferation, following ACTH and/or N-POMC(1-28) administration, has been previously demonstrated. In this study, we investigated the effect of ACTH and N-POMC(1-28) on the expression of adrenal cortex proteins related to cell cycle control such as cyclins D and P27(kip1). The administration of N-POMC upregulated cyclin D1 and D2 expression in the outer zone of the adrenal cortex; cyclin D3 expression was upregulated in the cortex inner zone even after administration of ACTH. Both ACTH and N-POMC peptides induced a decrease in the P27(kip1) expression in the ZG. These novel findings suggest that the POMC-derivate peptides, ACTH and N-POMC, promote proliferation in the adrenal cortex by upregulating the D2 and D3 cyclins and downregulating the P27(kip1) expression.

Desenvolvimento embrionário da carpa prateada (Hypophthalmichthys molitrix Valenciennes, 1844)

O estudo e o conhecimento dos primeiros dias de vida dos peixes são de extrema importância, especialmente para espécies selvagens e nativas, potencialmente viáveis para a piscicultura. Desta forma o trabalho teve como objetivo acompanhar o desenvolvimento embrionário da Hypophthalmichthys molitrix. A temperatura média da água apresentou pequena variação durante a realização do trabalho de 24,07 ± 0,23C, já os outros parâmetros como: pH, Alcalinidade, Dureza e Amônia, permaneceram constantes. A primeira segmentação foi observada com 22 minutos. A formação da blástula ocorreu com 2 horas e 27 minutos, com 9 horas e 12 minutos observou-se a gástrula com 90% de formação, o fechamento do blastóporo se deu com 9 horas e 49 minutos e a diferenciação do embrião com 12 horas e 56 minutos. A eclosão ocorreu com 20 horas e 3 minutos após a fertilização, totalizando desta forma 482,6 horas/graus. O desenvolvimento embrionário da carpa prateada Hypophthalmichthys molitrix ocorre dentro do período normal, como observado para as de mais espécies de peixes de água doce de regiões tropicais, mas muito ainda tem que ser estudado na área de embriologia de peixes, a fim de buscar maior conhecimento sobre o desenvolvimento destes animais.