FACTORS AFFECTING CHINESE HAMSTER OVARY CELL PROLIFERATION AND VIABILITY

Advantageous cultivation procedures for the Chinese hamster ovary (CHO) cells are necessary for the productive commercial production of biopharmaceuticals. A main challenge that needs to be addressed during the process development is the differences in each cell line requirements concerning the nutrients and feed strategies in order to achieve the desired growth characteristics. Therefore, within the current research, a naïve high cell density serum free suspension adapted CHO cell line was tested with glucose and glutamine rich feeds in fed-batch Erlenmeyer shake flask cultures. Glucose consumption rate was adjusted to develop the optimal feed strategies. Obtained results indicated that high glucose and l-glutamine feeding did not improve maximum viable cell density compared to the control samples. During the exponential phase, cell proliferation and viability of all feeds showed no statistically significant difference. Instead, the fed-batch processes tested led to statistically significant differences in viable cell density and cell viability during the decline phase, compared to control (batch) culture. The difference between glucose and glutamine feeding was indistinguishable, most probably due to the concentration imbalance with the rest of the nutrients in feed. The overall study presented a method to slow down the decrease in CHO cell proliferation and viability during the decline phase, instead of increasing the maximum cell density at the plateau phase.

Spatio-temporal dynamics of the common skate species complex: Evidence of increasing abundance

Aim: The critically endangered common skate species complex is a large‐bodied and long‐lived batoid, which has experienced local extirpations and population declines over the past century mainly due to overfishing. Due to its decline, fisheries management measures were introduced to prevent further decline and fragmentation of populations. For example, in 2009, a landings prohibition was introduced in the European Union, which banned the retention of common skate onboard commercial fishing vessels with captured individuals to be discarded. We aimed to explore the spatial and temporal population dynamics of the common skate species complex, against the backdrop of changes in fisheries management measures. Location: Northeast Atlantic Ocean. Methods: We used publicly available fishery‐independent trawl survey data from several regions of the Northeast Atlantic shelf to examine trends in incidence and abundance for the common skate species complex. We also constructed a species distribution model to identify changes in the spatio‐temporal distribution of the common skate. Results: A sustained increase in the common skate species complex was evident in several areas of its distribution. An increase was observed in five separate trawl surveys encompassing distinct regions of its distribution. Despite the observed increase, little evidence of recolonizing previously extirpated areas was evident. Main conclusions: The findings demonstrate the effectiveness of fisheries management measures in contributing to an increase in the common skate species complex. Such measures may also be effective if applied to numerous other batoid species currently threatened with extinction.<br/

We can work it out: an enactive look at cooperation

The past years have seen an increasing debate on cooperation and its unique human character. Philosophers and psychologists have proposed that cooperative activities are characterized by shared goals to which participants are committed through the ability to understand each other’s intentions. Despite its popularity, some serious issues arise with this approach to cooperation. First, one may challenge the assumption that high-level mental processes are necessary for engaging in acting cooperatively. If they are, then how do agents that do not possess such ability (preverbal children, or children with autism who are often claimed to be mind-blind) engage in cooperative exchanges, as the evidence suggests? Secondly, to define cooperation as the result of two de-contextualized minds reading each other’s intentions may fail to fully acknowledge the complexity of situated, interactional dynamics and the interplay of variables such as the participants’ relational and personal history and experience. In this paper we challenge such accounts of cooperation, calling for an embodied approach that sees cooperation not only as an individual attitude toward the other, but also as a property of interaction processes. Taking an enactive perspective, we argue that cooperation is an intrinsic part of any interaction, and that there can be cooperative interaction before complex communicative abilities are achieved. The issue then is not whether one is able or not to read the other’s intentions, but what it takes to participate in joint action. From this basic account, it should be possible to build up more complex forms of cooperation as needed. Addressing the study of cooperation in these terms may enhance our understanding of human social development, and foster our knowledge of different ways of engaging with others, as in the case of autism

Significance of mechanical loading in bone fracture healing, bone regeneration, and vascularization

In 1892, J.L. Wolff proposed that bone could respond to mechanical and biophysical stimuli as a dynamic organ. This theory presents a unique opportunity for investigations on bone and its potential to aid in tissue repair. Routine activities such as exercise or machinery application can exert mechanical loads on bone. Previous research has demonstrated that mechanical loading can affect the differentiation and development of mesenchymal tissue. However, the extent to which mechanical stimulation can help repair or generate bone tissue and the related mechanisms remain unclear. Four key cell types in bone tissue, including osteoblasts, osteoclasts, bone lining cells, and osteocytes, play critical roles in responding to mechanical stimuli, while other cell lineages such as myocytes, platelets, fibroblasts, endothelial cells, and chondrocytes also exhibit mechanosensitivity. Mechanical loading can regulate the biological functions of bone tissue through the mechanosensor of bone cells intraosseously, making it a potential target for fracture healing and bone regeneration. This review aims to clarify these issues and explain bone remodeling, structure dynamics, and mechano-transduction processes in response to mechanical loading. Loading of different magnitudes, frequencies, and types, such as dynamic versus static loads, are analyzed to determine the effects of mechanical stimulation on bone tissue structure and cellular function. Finally, the importance of vascularization in nutrient supply for bone healing and regeneration was further discussedSignificance of mechanical loading in bone fracture healing, bone regeneration, and vascularizationpublishedVersio

Efeito de três FLA2s, (BaTX-I, BaTX-II e BaFLA2), isoladas do veneno da serpente Bothrops atrox, sobre a funcionalidade de macrófagos J774A.1 em cultura

Dissertação apresentada ao programa de Pós-graduação em Biologia Experimental da Universidade Federal de Rondônia, para obtenção do título de Mestre.As fosfolipases A2 (FLA2) são enzimas que catalisam a hidrólise da ligação acil-éster de fosfolipídios de membranas celulares na posição sn-2, liberando ácidos graxos e lisofosfolipídios. Os produtos desta reação podem, subsequentemente, serem utilizados para a síntese de eicosanóides com funções importantes em reações inflamatórias. No presente estudo investigamos os efeitos in vitro de duas miotoxinas básicas, BaTX-I, uma Asp-49 cataliticamente ativa e BaTX-II, uma Lys-49 homóloga inativa, e de uma miotoxina ácida, BaFLA2, também uma Asp-49 ativa cataliticamente, isoladas do veneno da serpente Bothrops atrox, sobre a ativação de macrófagos J774 A.1. Em concentrações não citotóxicas, as três toxinas não afetaram a capacidade de adesão e descolamento dos macrófagos. Apenas a BaTX-II, enzimaticamente inativa, aumentou a fagocitose mediada pelo receptor de complemento. Entretanto, as três toxinas, BaTX-I, BaTX-II e BaFLA2, induziram aumento na produção de ânion superóxido em macrófagos. Somente a BaTX-II aumentou o volume lisossomal dos macrófagos após 15 minutos de incubação. Aos 30 minutos, as três fosfolipases aumentaram este parâmetro, o que não foi observado no período de 60 minutos. Além disso, as três miotoxinas, BaTX-I, BaTX-II e BaFLA2, induziram a formação de corpúsculos lipídicos em macrófagos submetidos e não submetidos á fagocitose. Este resultado pode sugerir a função dos corpúsculos lipídicos como um reservatório de mediadores inflamatórios. Ainda, as toxinas BaTX-I e BaFLA2, foram capazes de aumentar a produção de TNF-α pelos macrófagos no período de 4 horas. A BaTX-II não exerceu este efeito no tempo estudado. Tomados em conjunto, os dados mostraram que, a despeito das diferenças na atividade catalítica, as três toxinas induziram eventos inflamatórios e o caráter ácido ou básico destas enzimas não parece contribuir para estes efeitos

Mildronate, an inhibitor of carnitine biosynthesis, induces an increase in gamma-butyrobetaine contents and cardioprotection in isolated rat heart infarction

The inhibition of gamma-butyrobetaine (GBB) hydroxylase, a key enzyme in the biosynthesis of carnitine, contributes to lay ground for the cardioprotective mechanism of action of mildronate. By inhibiting the biosynthesis of carnitine, mildronate is supposed to induce the accumulation of GBB, a substrate of GBB hydroxylase. This study describes the changes in content of carnitine and GBB in rat plasma and heart tissues during long-term (28 days) treatment of mildronate [i.p. (intraperitoneal) 100 mg/kg/daily]. Obtained data show that in concert with a decrease in carnitine concentration, the administration of mildronate caused a significant increase in GBB concentration. We detected about a 5-fold increase in GBB contents in the plasma and brain and a 7-fold increase in the heart. In addition, we tested the cardioprotective effect of mildronate in isolated rat heart infarction model after 3, 7, and 14 days of administration. We found a statistically significant decrease in necrotic area of infarcted rat hearts after 14 days of treatment with mildronate. The cardioprotective effect of mildronate correlated with an increase in GBB contents. In conclusion, our study, for the first time, provides experimental evidence that the long-term administration of mildronate not only decreases free carnitine concentration, but also causes a significant increase in GBB concentration, which correlates with the cardioprotection of mildronate.publishersversionPeer reviewe

Leikkauksen jälkeisen haavadreenin hoitotyö: opetusvideo sairaanhoitajaopiskelijoille

Haavaimulla eli haavadreenin käytöllä voidaan edistää leikkaushaavan paranemista sekä ehkäistä mahdollisia komplikaatioita poistamalla eritteitä leikkausalueelta, joita leikkauksien jälkeen voi esiintyä. Haavadreenin käyttöön liittyy aina infektioriski, sillä dreeni on vierasesine. Sen vuoksi sairaanhoitajan on hyvin tärkeää osata aseptinen työskentelytapa haavadreeniä käsiteltäessä. Haavadreenin huolellinen hoito ja oikeaoppinen käsittely ovat keskeisiä tekijöitä infektion ehkäisemisessä ja potilaan toipumisprosessin tukemisessa. Opinnäytetyön tarkoituksena oli tuottaa laadukas opetusvideo Diakonia-ammattikorkeakoulun Porin-kampukselle toisen vuoden sairaanhoitajaopiskelijoille. Opinnäytetyön tavoitteena on lisätä postoperatiivista hoitotyötä opiskelevien sairaanhoitajaopiskelijoiden tietoa ja osaamista leikkauksen jälkeisen haavadreenin hoitotyöstä ja näin lisätä myös potilasturvallisuutta sekä edistää ammatillisen osaamisen kehittymistä. Opetusvideon avulla opiskelijat saavat käytännönläheistä ja visuaalista tukea haavadreenin hoitotyön opetteluun. Opinnäytetyö on kehittämispainotteinen. Kehittämistyön tuotoksena tehtiin opetusvideo, jota sairaanhoitajaopiskelijat voivat hyödyntää opiskelussaan. Opetusvideo tarjoaa selkeän ja havainnollistavan esityksen haavadreenin hoitotyön perusperiaatteista, aseptisista työtavoista ja mahdollisista haasteista, joita hoitotyössä saattaa kohdata. Asiasanat: dreeni, haavadreeni, opetusvideo, sairaanhoitajat, hoitotyö, haav

Time-dependent cytokine expression in bone of experimental animals after hydroxyapatite (Hap) implantation

Proinflammatory cytokines mediate bone loss around the implants in patients with peri-implant disease. However, there is no complete data about the expression of cytokines into the bone around the implants. The aim of this work was to investigate the distribution and appearance of inflammatory cytokines and anti-inflammatory proteins in the bone of jaw of experimental rabbits in different time periods after HAp implantation. Material was obtained from 8 rabbits in lower jaw 6 and 8 months after HAp implants were placed. Tissues were processed for immunohistochemical detection of tumor necrosis factor alfa (TNFα), Interleukin 1, 6, 8, 10 (IL-1, IL-6, IL-8, IL-10) and defensin 2. Results demonstrated practically unchanged expression of IL-6 and IL-10 between both - experimental and control side 6 months after implantation, while IL-1 and IL-8 notably increased in control side. IL-1 and IL-10 expression did not change in either the experimental side nor the controle side after 8 months HAP implantation, but IL-6 and IL-8 demonstrated a decrease in the control sites. Only IL-8 was elevated with time in experimental sites, while IL-10 showed individual variations in 2 cases.publishersversionPeer reviewe