bluuebunny/crossref_metadata_2025_split huggingface dataset:
K562. A Human Cellular System Capable of Undergoing In Vitro Differentiation: Measurement of Genotoxic Parameters Useful for Cytotoxicity Evaluation
Authors: Sapora Orazio , Maggi Antonella , Maione Barbara , Pazzaglia Simonetta , Tabocchini Maria-Antonella 
Published: August 2020

The initial damage produced by ionising radiation and its subsequent repair have been studied in a cellular system. K562 cells are used which are capable of undergoing in vitro pseudoerythroid differentiation in the presence of 2mM butyric acid. The level of initial damage is similar in actively growing and 48-hour differentiated cells, while it is lower by a factor of two in cells irradiated after exposure to the inducer for 72 hours. In differentiated cells, the kinetics of repair measured up to 60 minutes is slower than that in actively growing cells. These findings suggest that the genome of actively proliferating cells is not only more susceptible to radiation-induced damage, but also more accessible to repair enzymes than the more compact genome of differentiated cells. Repair after a longer time interval has also been investigated. After 24 hours of repair, the amount of residual damage is higher in actively proliferating cells than in differentiated ones. However, in proliferating cells, DNA synthesis can interfere with repair of the lesions or vice versa, while in differentiated cells, due to the lack of proliferation, damage occurring in non-transcribing genes can presumably be sustained for longer periods without biological consequences. A method to evaluate the relative sensitivity of non-duplicating differentiated cells is proposed. It is based on measurement of the residual damage, detected after 24 hours of repair, and on the assumption that a relationship exists between unrepaired DNA damage (residual damage) and the ability of the cell to survive.
Evaluation of the Cytotoxicity of the First 20 MEIC Chemicals in Two Hepatoma Cell Lines with Different Xenobiotic Metabolism Capacities
Authors: Bassi Anna Maria , Bosco Ornella , Brenci Sabrina , Adamo Daniela , Penco Susanna , Piana Sandra , Ferro Margherita , Nanni Giorgio 
Published: August 2020

The cytotoxicity of the first 20 chemicals on the MEIC list was evaluated using two hepatoma cell lines having different xenobiotic metabolic capacities, one derived from the rat (HTC), and one of human origin (Hep G2). Two endpoints were measured to evaluate cytotoxicity: colony-forming ability (CF), and cell viability of the attached monolayer (CV), evaluated as total macromolecular content. The CF test provided the most sensitive endpoint, due to the lower number of exposed cells in comparison with the CV test. Using the CF assay, the Hep G2 cell line showed higher sensitivity than the HTC cell line to some chemicals known to be metabolised in vivo. The IC50s obtained under the different experimental conditions varied as a consequence of phenotypic differences between the cell lines, and of the nature of the endpoints. The most toxic chemicals were malathion, diazepam and amitriptyline (IC50: 0.001-0.1mM). The solvents tested produced the lowest toxic effects (IC50: > 10mM). These findings suggest that hepatoma cell lines possessing various specific enzyme activities could be usefully employed in a battery of tests designed to reproduce in vitro the wide range of biochemical properties expressed by the cells in the whole organism.
A European Interlaboratory Evaluation Study of an In Vitro Ocular Irritation Model (Skin2™ Model ZK1100) using 15 Chemicals and 3 Shampoos
Authors: Pirovano Raffaella , Zaninelli Pietro , Noben Jos , Logemann Pamela , Southee Jacqueline , Joller Peter , Coquette Alain 
Published: August 2020

The aim of this interlaboratory study was to ascertain the feasibility of the use of the in vitro Skin2™ Model ZK1100 kit as a tissue substrate for cytotoxicity studies. Eighteen compounds — 15 raw chemicals and 3 finished products (commercial shampoos) — were tested in four different laboratories on two separate occasions, and the data obtained were compared with the data available in the literature on eye irritancy potential. The kit was easy to manage, and no technical difficulties were encountered in the estimation of MTT reduction, LDH and PGE2 release in the medium. Similar MTT50 values were recorded in two independent experiments in each laboratory, and the rankings of irritation potency within the same chemical class were comparable. Both LDH and PGE2 were important parameters to be taken into account along with MTT50 values in establishing the irritation potential of some categories of compounds.
Evaluation of the First Ten MEIC Chemicals in the Allium Test
Authors: Fiskesjö Geirid , Levan Albert 
Published: August 2020

After joining the MEIC (multicentre evaluation of in vitro cytotoxicity) programme, and submitting the first ten compounds of the programme to the Allium test, we found that this test correlates well with several other tests (e.g. MIT-24 cell test, and tests with mice, rats or humans in vivo). The compounds tested were paracetamol, acetylsalicylic acid, iron sulphate, diazepam, amitriptyline, digoxin, ethylene glycol, methanol, ethanol and isopropanol. Among them, paracetamol turned Out to induce chromosome breakage in the meristems of Allium cepa root tips. This is the same qualitative response to paracetamol as that obtained previously in human lymphocytes in vivo and in vitro. The present results indicate that the eukaryotic higher plants may serve as highly useful test systems for biological risk evaluation.
The Use of Several Leakage Markers which Enable Platelet Membrane Damage and Platelet Activation to be Distinguished
Authors: Mäkelä Jan-Henrik , Isomaa Boris 
Published: August 2020

We have studied the effects of four differently charged detergents and sphingosine on platelet membrane integrity and platelet activation, using the leakage of pre-loaded 43K+, pre-loaded [14C]-serotonin ([14C]-5-HT) and lactate dehydrogenase (LDH) as markers. Sphingosine, at a concentration of 20μM, induced a release of [14C]-5-HT as well as 43K+, without releasing LDH. The [14C]-5-HT743K+ ratio was about 1.5. A similar leakage pattern and the same [14C]-5-HT/43K+ ratio was also induced by thrombin, a potent platelet agonist, indicating that the release induced by sphingosine is due to platelet activation and not to membrane damage. The detergents, on the other hand, induced a marked release of 43K+ followed by an increased release of LDH. No pronounced leakage of [14C]-5-HT occured with the detergents; the [14C]-5-HT/43K+ ratio was ≤ 0.2. In this manner, we can distinguish between marker-release caused by membrane-damage and that caused by platelet-activation, by using the combination of leakage markers presented here. Our study also showed that fixation prior to centrifugation is crucial in order to avoid leakage of markers, which are released upon activation of the platelets. About 35% of the [14C]-5-HT content of the platelets was released when unfixed platelets were centrifuged, and, interestingly, the detergents, as well as ethanol, inhibited the centrifugation-induced release of [14C]-5-HT.
Critical Evaluation of an Agarose Diffusion Method for the Assessment of Eye Irritancy
Authors: Cottin Martine , Zanvit Alain , Faquet Brigitte , Rougier André , Dossou Koovi G. 
Published: August 2020

The agarose overlay method is often used to study the cytotoxicity of non-liquid products. However, diffusion through agarose is a limiting factor in some cases, and this reduces contact between the cells and the test material. We have developed a new model aimed at reducing this phenomenon. In our technique, rather than being cultured in a monolayer, V79 cells are suspended in 1% agarose medium. We obtained a good linear correlation between this method and the usual L-929 cell monolayer technique (r = 0.95; n = 31). In addition, the new technique is more sensitive. In particular, the cytotoxicity of surfactants is apparent at lower concentrations. Using this technique, we studied 115 miscellaneous raw materials and cosmetic products, for which in vivo historical ocular irritancy data were available. As expected, our method was unsuitable for the study of alkaline and acidic products, as well as water-alcohol-based lotions. We also adapted the method for testing mascaras and obtained a good ranking for eye irritancy. For the other 93 products (surfactants, shampoos, lotions, gels, emulsions and miscellaneous products), the mean diameter of the lysis zone after contact for 18 hours was used as the endpoint for comparison with in vivo maximum average scores. There was a strong linear/rank correlation between the two parameters (r = 0.83/rho = 0.79). Furthermore, the concordance between the in vivo and in vitro data in a three-class ranking system was good (weighed kappa index = 0.81). These results suggest that the agarose diffusion method is a potentially useful screening tool for the ocular safety of the categories of products described above.
The Galileo Data Bank on Toxicity Testing with In Vitro Alternative Methods. II. Toxicology Profiles of 20 Chemicals
Authors: Loprieno Nicola , Boncristiani Guido , Bosco Elena , Nieri Maria , Loprieno Gregorio 
Published: August 2020

The identification of the hazard of chemicals to man has relied on the use of several animal models. However, the availability of various cell toxicity models as alternatives to the use of animals has stimulated attempts to evaluate in vitro data for use in the prediction of human toxicity. The cell toxicity models developed previously are capable of indicating a variety of endpoints susceptible to the activity of various chemical substances. The in vitro data derived so far from testing a variety of types of chemicals, have been used to develop toxicology profiles for twenty chemicals, which are presented in this paper. Data have been selected from among those already entered in the Galileo Data Bank, a computerised data system containing all the available existing data derived using in vitro methods.
Species-specific Effects of Nitromethylene Heterocycle Insecticides on Nicotinic Receptors in Locust Neurons and Mouse N1E-115 and BC3H1 Cells
Authors: Oortgiesen Marga , Zwart Ruud , Vijverberg Henk P.M. 
Published: August 2020

The effects of nitromethylene heterocycle (NMH) insecticides on subtypes of nicotinic acetylcholine (nACh) receptors were investigated in locust thoracic ganglion neurons, mouse N1E-115 neuroblastoma cells, and mouse BC3H1 muscle cells by using electrophysiological techniques. In locust neurons, all of the six NMH insecticides tested induced transient inward currents resembling nicotinic ACh-induced inward currents, while, in the continued presence of the NMH compounds, the ACh-induced inward current was blocked. The amplitude of the inward current and the blocking effects of the NMH insecticides were enhanced by concentrations between 0.1 and 10μM. Cross-desensitisation with the ACh-induced inward current confirmed that the NMH-induced inward current was governed by the activation of nACh receptors. Mammalian endplate type nACh receptors in BC3H1 cells and mammalian neuronal type nACh receptors in N1E-115 cells were much less sensitive to the NMH insecticides than the locust neuronal nACh receptors. At a concentration of 10μM, which blocked 80–100% of the ACh-induced inward current in locust neurons, NMH insecticides only partially blocked the ACh-induced inward currents mediated by the two subtypes of mammalian nACh receptors. NMH insecticides also failed to induce significant agonist effects in the mammalian cells at this concentration. The results provide a possible explanation for the selectively greater toxicity of NMH insecticides to insects than to vertebrates, at the level of nACh receptor subtypes and, hence, demonstrate that this in vitro approach is valuable for the investigation of species-specific interactions of compounds at their target site.
Why is it Unethical to use Chimpanzees in the Laboratory?
Authors: Goodall Jane 
Published: August 2020

Chimpanzees are more like humans than any other living beings, differing in the composition of their DNA by just over one per cent. There are striking similarities in the anatomy and wiring of the chimpanzee and human brains and central nervous systems. Thus, it should not be surprising to find that there are also striking similarities in the social behaviour, emotional needs and expressions, and cognitive abilities of chimpanzees and humans. These similarities have become increasingly apparent during the last 15 years. Chimpanzees in the wild develop close affectionate bonds between family members that may persist throughout their lifetime of 50 years or more, and examples of true altruism, when individuals protect or even save the lives of non-related companions. Chimpanzees use many objects as tools, and tool-using behaviours differ from place to place across their range. Indeed, there are a number of behaviours that vary between different groups — evidence of cultural traditions passed from one generation to the next through observational learning and imitation. Thus, chimpanzees have a very special relationship with humans. A healthy adult chimpanzee is more similar to a healthy adult human in the expression of the intellect than a brain-damaged human, yet in many medical research facilities, chimpanzees are maintained in bleak, bare cages measuring only 5’ x 5’ x 7'. They may remain in these prisons for life. We do not treat hardened human killers so badly in our society today—there would be a public outcry if we did. I feel strongly that the use of a being so like us, as a human guinea-pig, is not morally justified, and to that end the Jane Goodall Institute has been involved in three workshops with the aim of clarifying the extent to which they are seen to be useful in diseases such as hepatitis and AIDS research. There is no consensus among scientists regarding their usefulness at the present time. If the proposed experiments of transplanting chimpanzee bone marrow tissue into AIDS patients go ahead in the Netherlands, it will be a sad blow for chimpanzee liberation. The attitude of those who believe that any use of non-human primates can be justified provided it results in some benefit, or expected benefit, to humankind, is of precisely the same mind set as that which once allowed us to exploit human beings of another race and use them as slaves. Once we admit that chimpanzees have minds and feelings, are capable of sadness, fear and despair, are able to feel pain, show altruism, and are capable of communicating with each other and with humans in a man-made language, we have to ask serious questions, initially of ourselves, as to whether we should continue to use them in medical research.
Alternatives in Canada
Authors: Griffin Gilly 
Published: August 2020

In Canada, promotion of the Three Rs concept of alternatives occurs largely through the Canadian Council on Animal Care. Institution-based animal care committees are required to conduct an ethical review of all protocols which use animals in research, teaching or testing. Investigators are expected to use animals only when their best efforts to find an alternative have failed. Those using animals are required to employ the most humane methods on the smallest number of animals possible. The Joseph F. Morgan Research Foundation was established to promote the development and use of alternative methods in Canada. The focus of the Foundation has been on the use of alternative methods for testing purposes. However, the Foundation also encourages the acceptance of each of the Three Rs throughout Canadian science. The Foundation is therefore in the process of developing a refinement alternatives programme.
The Three Rs in Research and Education: A Long Road Ahead in the United States
Authors: Orlans F. Barbara 
Published: August 2020

Attitudes toward the Three Rs concept of refinement, reduction and replacement in the United States in research and education are widely divergent. Positive responses have come from several sources, notably from four centres established to disseminate information about alternatives. Funding sources to support work in the Three Rs have proliferated. The activities of institutional oversight committees have resulted in the nationwide implementation of important refinements. In the field of education, student projects involving pain or death for sentient animals have declined, and the right of students to object to participation in animal experiments on ethical grounds has been widely established. However, there is still a long way to go. Resistance to alternatives is deep-seated within several of the scientific disciplines most closely associated with animal research. The response of the National Institutes of Health to potentially important Congressional directives on the Three Rs has been unsatisfactory. The prestigious National Association of Biology Teachers, which at first endorsed the use of alternatives in education, later rescinded this policy, because of opposition to it. An impediment to progress is the extreme polarisation of viewpoints between the biomedical community and the animal protectionists.
A cross-sectional gender-sensitive analysis of depressive symptoms in patients with advanced cancer
Authors: Shapiro Gilla K , Mah Kenneth , de Vries Froukje , Li Madeline , Zimmermann Camilla , Hales Sarah , Rodin Gary 
Published: August 2020

Background: Patients with advanced cancer commonly report depressive symptoms. Examinations of gender differences in depressive symptoms in patients with advanced cancer have yielded inconsistent findings. Aim: The objective of this study was to investigate whether the severity and correlates of depressive symptoms differ by gender in patients with advanced cancer. Design: Participants completed measures assessing sociodemographic and medical characteristics, disease burden, and psychosocial factors. Depressive symptoms were examined using the Patient Health Questionnaire, and other measures included physical functioning, symptom burden, general anxiety, death related distress, and dimensions of demoralization. A cross-sectional analysis examined the univariate and multivariate relationships between gender and depressive symptoms, while controlling for important covariates in multivariate analyses. Setting/participants: Patients with advanced cancer ( N = 305, 40% males and 60% females) were recruited for a psychotherapy trial from outpatient oncology clinics at a comprehensive cancer center in Canada. Results: Severity of depressive symptoms was similar for males ( M = 7.09, SD = 4.59) and females ( M = 7.66, SD = 5.01), t(303) = 1.01, p = 0.314. Greater general anxiety and number of cancer symptoms were associated with depressive symptoms in both males and females. Feeling like a failure ( β = 0.192), less death anxiety ( β = –0.188), severity of cancer symptoms ( β = 0.166), and older age ( β = 0.161) were associated with depressive symptoms only in males, while disheartenment ( β = 0.216) and worse physical functioning ( β = 0.275), were associated with depressive symptoms only in females. Conclusions: Males and females report similar levels of depressive symptoms but the pathways to depression may differ by gender. These differences suggest the potential for gender-based preventive and therapeutic interventions in this population.
NLRC5 alleviated OGD/R-induced PC12-cell injury by inhibiting activation of the TLR4/MyD88/NF-κB pathway
Authors: Zhang Zhen , Sun Yuhan , Chen Xin 
Published: August 2020

Objective To assess the role of NOD-like receptor C5 (NLRC5; a major NLRC family protein that regulates immunity, inflammation and tissue fibrosis), in cerebral ischemia-reperfusion injury, characterized by inflammation and oxidative damage. Methods Blood NLRC5 levels were assessed in neonates with cerebral ischemia and in healthy controls. A stable PC12 cell line was established that overexpressed or knocked down NLRC5. Inflammatory responses, apoptosis rate and oxidative damage in PC12 cells under oxygen-glucose deprivation/reperfusion (OGD/R) conditions were evaluated using enzyme-linked immunosorbent assay (ELISA), terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) and reactive oxygen species (ROS) assay. Results Blood NLRC5 levels were suppressed in neonates with cerebral ischemia. ELISAs showed that NLRC5 suppressed levels of tumour necrosis factor-α, interleukin (IL)-6, IL-1β, ROS and superoxide dismutase in OGD/R-treated PC12 cells. Furthermore, NLRC5 overexpression was associated with reduced apoptosis rate in PC12 cells treated by OGD/R. Overexpression of NLRC5 also inhibited levels of toll-like receptor (TLR)4, myeloid differentiation primary response protein MyD88 (MyD88) and phosphorylated nuclear factor kappa B-transcription factor p65 (NF-κB p-p65) in PC12 cells, and decreased nuclear levels of NF-κB p-p65. Conclusion NLRC5 alleviated inflammatory responses, oxidative damage and apoptosis in PC12 cells under OGD/R conditions by suppressing activation of the TLR4/MyD88/NF-κB pathway.
Prediction of severe ovarian hyperstimulation syndrome in women undergoing in vitro fertilization using estradiol levels, collected ova, and number of follicles
Authors: Madrazo Ivan , Vélez Monserrat Fabiola , Hidalgo Josue Jonathan , Ortiz Ginna , Suárez Juan José , Porchia Leonardo M. , Gonzalez-Mejia M. Elba , López-Bayghen Esther 
Published: August 2020

Objective Our objective was to determine whether estradiol (E2) levels (Day 3 and fold change to Day 10), antral follicle count (AFC), and number of ova collected could predict ovarian hyperstimulation syndrome (OHSS) and culdocentesis intervention. Methods We conducted a retrospective review of patient charts between January 2008 and December 2017. OHSS was defined using American Society for Reproductive Medicine criteria. Predictability was evaluated by measuring the area under the receiver operating characteristic curve (AUC). Results The cohort included 319 women (166 controls, 153 OHSS, of whom 54 had severe OHSS). The OHSS group had higher E2Day 3 (249 ± 177 vs. 150 ± 230 ng/L), E2FoldChange (32.2 ± 29.1 vs. 20.1 ± 23.8), AFC (18.2 ± 9.1 vs. 11.6 ± 8.3), and number of ova collected (21.1 ± 9.0 vs. 10.1 ± 6.5). E2Day 3 (AUC = 0.76, 95%CI: 0.71–0.82), E2FoldChange (AUC = 0.71, 95%CI: 0.65–0.77), AFC (AUC = 0.75, 95%CI: 0.70–0.81), and number of ova collected (AUC = 0.85, 95%CI: 0.81–0.89) were predictive for OHSS. All variables were predictive for culdocentesis intervention (E2Day 3: AUC = 0.63, 95%CI: 0.55–0.70; E2FoldChange: AUC = 0.63, 95%CI: 0.55–0.71; AFC: AUC = 0.74, 95%CI: 0.68–0.80; number of ova collected: AUC = 0.80, 95%CI: 0.75–0.85). Conclusions Day 3 E2 levels and number of ova collected predict patients who could develop OHSS and may require culdocentesis.