bluuebunny/crossref_metadata_2025 huggingface dataset:
The knowing world: A new global history of science
Authors: Delbourgo James
Published: April 2019

This article proposes a new global approach to the history of science centered on questions of geopolitics, historical consciousness, and cultural identity. Arguing that the field is now at a crossroads between its longstanding focus on the history of the natural sciences in the Western world, and the prospect of some form of worldwide history of science, the article describes a new undergraduate lecture course, designed by the author and taught at Rutgers and Harvard since 2015, which neither begins in Western Europe nor culminates with the United States. It aims to articulate an original vision for the field on this basis. Histories of science can and should offer deep histories of the global present, it is argued, by rethinking how historical narratives involve geographical decisions about where to focus analytical attention (and where not) and tackling narrative and geography together as linked issues. The approach proposed here is neither science in context nor knowledge in transit but engages the notion of a knowing world: one made up of multiple scientific cultures and long historical memory, and requiring dialectical movement back and forth across both space and time on a worldwide scale to grasp the scientific past’s importance for the present, as well as the present’s impact on how we perceive the past. Explicitly addressing polemics of identity, culture, race, and nationhood can help us to construct a more civic-minded and geopolitically informed history of science of use in the changing circumstances of the twenty-first century.
The Evolution of Aging With Pride—National Health, Aging, and Sexuality/Gender Study: Illuminating the Iridescent Life Course of LGBTQ Adults Aged 80 Years and Older in the United States
Authors: Fredriksen Goldsen Karen , Kim Hyun-Jun , Jung Hyunzee , Goldsen Jayn
Published: April 2019

Aging with Pride: National Health, Aging, and Sexuality/Gender Study is the first federally funded study addressing aging among LGBTQ older adults throughout the United States. This article examines the evolution of this landmark study and explores the well-being of LGBTQ adults aged 80 years and older ( n = 200), the most underrepresented group in the field. Based on the Iridescent Life Course, we examined the diverse, intersectional nature of LGBTQ older adults’ lives, finding high levels of education and poverty. Microaggressions were negatively associated with quality-of-life and positively associated with poor physical and mental health; the inverse relationship was found with mastery. When the oldest encountered risks, it resulted in greater vulnerability. This longitudinal study is assessing trajectories in aging over time using qualitative, quantitative, and biological data and testing evidence-based culturally responsive interventions for LGBTQ older adults. Research with LGBTQ oldest adults is much needed before their stories are lost to time.
Effects of Functional Discrimination Training on Initial Receptive Language in Individuals With Autism Spectrum Disorder
Authors: Eldevik Sigmund , Aarlie Hege , Titlestad Kristine B. , Kazemi Ellie , Elsky Greg
Published: April 2019

The success of behavior-analytic procedures to teach language to individuals with autism spectrum disorder (ASD) has been well established in the literature. Nevertheless, some individuals may not learn any receptive or expressive language following extensive teaching efforts. We examined the effects of two reinforcement contingencies, functional and arbitrary, on increasing the level of auditory–visual conditional discriminations in children with ASD with a history of having difficulty learning discriminations. We evaluated the effects of the reinforcement contingencies by comparing the number of trials needed to establish discriminations in an adapted alternating treatment design. We found that five out of eight participants showed more rapid acquisition and demonstrated discrimination between more items in the functional reinforcement condition. The remaining three participants did not exhibit any discrimination in either condition within the allotted 500 trials/20 days. These findings suggest that using functional reinforcement procedures may be a helpful alternative for individuals who do not learn discriminations through traditional procedures.
Kimura Disease: An Unusual Cause of Head and Neck Masses. Report of 2 Cases
Authors: Viswanatha Borlingegowda
Published: April 2019

Kimura disease is an uncommon chronic, benign, inflammatory condition of unknown etiology. It occurs endemically in Asia and sporadically in the West. Because it often imitates neoplastic or inflammatory processes of the head and neck, otolaryngologists and head and neck surgeons should be aware of its clinical presentations. In this article, the author reports 2 cases of Kimura disease—1 that was characterized by an unusual bilateral presentation and 1 that was managed conservatively—and he briefly reviews the literature on this entity.
Disseminated Plasmacytoma of the Thyroid
Authors: Park Brian J. , Kalish Randy J. , Vercillo Arthur P.
Published: April 2019

Extramedullary plasmacytomas most commonly occur in the nasal cavity, nasopharynx, paranasal sinuses, and larynx. Thyroid involvement is rare, as fewer than 75 cases have been previously reported in the literature. We report a new case of disseminated plasmacytoma of the thyroid, which occurred in a 68-year-old woman. The diagnosis was made with an incisional biopsy. The patient initially experienced a complete response to radiotherapy (46.8Gy in 26 fractions), but she developed multiple metastases at distant sites, including the breast, abdominal wall, and buttock despite various chemotherapy regimens. Ultimately, she was placed on palliative radiotherapy. We also briefly review the diagnostic and treatment options for patients with extramedullary plasmacytoma.
Correlation between the Voice Handicap Index and Voice Laboratory Measurements after Phonosurgery
Authors: Cheng Jeffrey , Woo Peak
Published: April 2019

Phonosurgery is an effective treatment for some vocal fold pathologies, and the Voice Handicap Index (VHI) survey has been shown to be a useful instrument for evaluating treatment effectiveness. We conducted a nonrandomized, prospective study of 21 patients who underwent phonosurgery for the treatment of non-neoplastic vocal fold lesions at our academic tertiary-care referral center. Our goals were to compare pre- and postoperative VHI scores (subjective assessments) and pre- and postoperative results of acoustic and aerodynamic tests (objective assessments). We sought to determine if there was any correlation between the subjective and objective findings. We looked for differences between professional voice users (n = 10) and nonprofessional voice users (n = 11) in both subjective and objective measures. We found statistically significant differences between pre- and postoperative values in three of four VHI parameters, but in only one of 13 objective measures. There was no correlation between preoperative VHI scores and preoperative acoustic and aerodynamic test results. The professional voice users expressed greater postoperative improvement as reflected by lower VHI scores than did the nonprofessional voice users, confirming that the former are more negatively affected by a voice disability.
A Rare Cause of Stapedotomy Failure: When the Depth Marker Becomes a Locking Device: A Report on 2 Cases
Authors: Daval Mary , Guevara Nicolas , Alotaibi Naif , Williams Marc , Ayache Denis
Published: April 2019

We report 2 cases of stapes surgery failure due to the depth marker groove of the piston of a Big Easy titanium prosthesis getting caught at the edge of the stapedotomy hole. In the first case, the stapedotomy failure, detected peroperatively, was corrected by modifying the angulation of the shaft of the prosthesis. In the second case, the failure occurred after a delay during which scar tissue developed around the depth marker groove, hampering the movement of the prosthesis. The usual postoperative imaging did not detect this complication. The diagnosis was made during revision surgery and the condition was treated by enlarging the hole in the footplate. Alternatively, the use of a piston without a depth marker groove would have resolved the problem. Although a rare event, the possibility of stapes surgery failure due to the depth marker groove should not be overlooked.
Chemotherapy-Induced First Bite Syndrome: A Case Report in a Patient With Hodgkin Lymphoma
Authors: Valenzuela Carla V. , Bartlett Nancy L. , Bradley Joseph P.
Published: April 2019

Vinca alkaloids are known to cause bilateral jaw pain that occurs once during the chemotherapy course. We report a patient with first bite syndrome (FBS) during active treatment with chemotherapy. A patient with Hodgkin lymphoma presented with unilateral jaw pain after beginning his chemotherapy regimen. Pain was worse with the first bite of each meal and dissipated over subsequent bites. Workup was negative for any lesions in the parotid, parapharyngeal space, or infratemporal fossa. Pain was timed closely with chemotherapy administration and would improve prior to next cycle. A trial of botulinum chemodenervation failed to completely relieve symptoms. The patient noted resolution of symptoms after the completion of chemotherapy. We report a case of FBS, which may represent the jaw pain seen commonly with administration of vinca alkaloids. There appears to be a correlation between onset and duration of first bite symptoms with chemotherapy administration.
Does Everything Fall Apart? Life Assessments Following a Gray Divorce
Authors: Crowley Jocelyn Elise
Published: April 2019

The divorce–stress–adjustment theoretical framework emphasizes not only the negative consequences associated with marital dissolution but also the positive outcomes that can eventually emerge after former spouses part ways. This study aims to examine the subjective life assessments of the heterosexual gray divorced population within the first 3 years of their marital splits with respect to this framework. Qualitative data based on 66 in-depth interviews of the gray divorced population were collected and analyzed using grounded theory methods. Negative consequences of gray divorce included financial worries and loneliness. However, participants also identified positive aspects of their new lives, including higher levels of overall happiness, liberation from their ex-spouses, and enhanced independence and freedom. These results show that immediately after their marriages dissolve, members of the gray divorced population experience complex reactions in accordance with the divorce-stress-adjustment framework. More specifically, these men and women see both challenges and opportunities after their gray divorces.
Patterns of Immigration and Public Assistance Utilization, 1970–1980
Authors: Jensen Leif
Published: April 2019

The recent surge in immigration to the United States has rekindled debate over the economic burden imposed by immigrants. This article explores the utilization of public assistance by immigrants and natives. The analysis goes beyond existing research by utilizing repeated cross-sectional data. Descriptive tables show that despite their higher poverty rates, immigrant families had only minimally higher public assistance recipiency rates compared to natives. Multivariate logistic regression analyses reveal that immigrant families were generally less likely than natives to receive public assistance, other things equal. There were exceptions to this generalization according to year of observation and race/ethnicity.
The Institutional Structure of Immigration as a Determinant of Inter-Racial Competition: A Comparison of Britain and Canada
Authors: Reitz Jeffrey G.
Published: April 2019

This study attempts to explain differences between two countries—Britain and Canada—in the reception given to immigrant racial minorities in the post-war years. It is suggested that among the factors relevant to this explanation, importance should be attributed to cross-national differences in the institutional structure of immigration itself. Supporting comparative evidence comes from published research, public opinion polls, newspaper reports, parliamentary debates and the legislative records.
PART III: Migration, Politics, and Ethics: Realistic and Idealistic Approaches to the Ethics of Migration
Authors: Carens Joseph H.
Published: April 2019

There are two approaches to morality. The realistic approach wants to avoid too large a gap between the ought and the is and focuses on what it is possible given existing realities. This approach, however, inhibits us from challenging fundamentally unjust institutions and policies. The idealistic approach, in contrast, requires us to assess current reality in light of our highest ideals. Its weakness is that it may not help us answer the question of how to act in this non-ideal world. Discussions about the ethics of migration require a full range of perspectives using both approaches.
PART IV: Rights of Migrants, and Rights of Political Communities: Cultural Minority Rights for Immigrants
Authors: Bauböck Rainer
Published: April 2019

This paper argues that cultural minorities enjoy a basic right to recognition and rejects the idea that migrants implicitly renounce their cultural claims when they leave their countries of origin, when they enter the receiving society, or when they may return. Need for cultural recognition arises from the “ethnicization” of immigrant minorities. A catalog of minority rights provides a reference list for assessing the particular cultural rights of immigrant minorities. They can generally claim rights which recognize a multicultural transformation of receiving societies (equality, liberty, protection from individual discrimination, and public resources for cultural reproduction). However, most immigrants cannot reasonably claim rights derived from historic discrimination, or from collective autonomy within a federation of potentially separate polities.
Mass Exoduses: The Response of the United States
Authors: Churgin Michael J.
Published: April 2019

The focus of this article is on mass immigration to the United States and the country's response to various groups of immigrants. After presenting historical background dating back to the pre-Civil War era, attention is given to the Cuban and Haitian mass movements of recent years and to the refugees coming from Vietnam, the former Soviet Union, and Latin American countries. The article concludes that the United States has utilized international agreements regarding the settlement of large numbers of people only when they facilitate government action.
A Comparison of Three Cytotoxicity Endpoints in the Corneal HCE-T Model
Authors: Clothier Richard , Orme Alex , Walker Tracey L. , Ward Sherry L. , Kruszewski Francis H. , DiPasquale Louis C. , Broadhead Caren L.
Published: April 2019

The prediction of ocular irritation potential from in vitro assays still presents a problem, despite a number of validation trials. A study with coded cosmetic formulations, for which historic in vivo data were available, has been conducted with a human corneal multilayered model system. This corneal model, the HCE-T model, was developed by using HCE-T cells, a transfected human corneal epithelial cell line. The relative effectiveness of three endpoints that provide a measure of cytotoxicity in the HCE-T model was evaluated. Cell viability immediately after exposure to the test materials was determined by using the MTT and Alamar Blue™ (AB) assays, and, 24 hours later, by using the MTT, AB and lactate assays. Viability measurements with the MTT, AB and lactate assays gave similar dose–response curves at the 24-hour endpoint. One formulation (an anti-dandruff shampoo) caused a less severe drop in viability in assays conducted immediately after the exposure than at the 24-hour time-point. There was little deterioration in viability with the other test materials. The ranking of the test formulations on the basis of relative loss of viability and release of lactate resulted in the same order as for the Modified Maximum Average Draize Test Score. Comparison of the HCE-T model cytotoxicity assay results with historic in vitro data from two different cytotoxicity assays, conducted by using fibroblast monolayer cultures and the same materials, indicated that the multilayered corneal model had a greater predictive ability. The results of a blind trial with the lactate assay in two laboratories indicated that the techniques required were transferable between laboratories. The lactate results were reproducible between laboratories, even when cultures derived from different passage human corneal cells were tested, provided that the passage number was below 20.
Methyl Green-Pyronine Staining of Porcine Organotypic Skin Explant Cultures: An Alternative Model for Screening for Skin Irritants
Authors: Jacobs John J.L. , Lehé Cynthia , Cammans Keith D.A. , Das Pranab K. , Elliott Graham R.
Published: April 2019

We describe a new alternative method for screening for skin irritants by using fresh intact porcine skin biopsies. Test chemicals were applied to the epidermis of the biopsies, which were then incubated for different times in tissue culture medium at 37°C and with 5% carbon dioxide. A decrease in epidermal keratinocyte RNA staining, visualised in frozen sections by using a modified methyl-green pyronine (MGP) staining procedure, was employed as a marker of irritancy. If a decrease in epidermal RNA was observed after incubation for 4 hours (strong irritant), the chemical had an MGP score of 3; if after incubation for 24 hours (moderate irritant), the MGP score was 2; and if after incubation for 48 hours (weak irritant), the MGP score was 1. If no keratinocyte cytotoxicity was observed after incubation for 48 hours, the chemical was classified as non-irritant (MGP score = 0). At least three ears were used per chemical. The average MGP score was used to classify the chemical. Based on the MGP score for 20% sodium dodecyl sulphate, chemicals classified as strong or moderate irritants by using the MGP test were grouped together as category R38 chemicals. Weak irritants or non-irritants were not classified (NC). The MGP staining correctly identified 23 of 25 skin irritants for which reference data were available.
The High Production Volume Chemical Challenge Program: The Rodent LD50 and its Possible Replacement
Authors: Rosenkranz Herbert S. , Cunningham Albert R.
Published: April 2019

The High Production Volume Chemical Challenge Program provides an opportunity to re-examine the usefulness and informational value of tests currently used to obtain preliminary hazard identification data. With a view to assessing the mechanistic information provided by the rodent LD50 test and to ascertain the possibility of replacing it with other “more acceptable” assays, we used a recently developed approach to determine the relationship of the LD50 assay to other toxicological protocols. Our analyses indicate that, of the assays examined, the LD50 was significantly related to toxicity in cultured cells and to binding at the Ah receptor.
The Peterborough Hospital Human Tissue Bank
Authors: Womack Christopher , Gray Neil , Aikens Julie , Jack Alison
Published: April 2019

The Peterborough Hospital Human Tissue Bank, based in the Cellular Pathology Department of the District Hospital, has been successful in supplying commercial biomedical companies with human tissue for research purposes. Tissue is obtained from routine surgical specimens sent to the laboratory for diagnostic testing and from cadaveric donors examined in the hospital mortuary. All tissue is obtained legally and with the full informed consent of the patient, donor or relative, as appropriate. The mechanism of retrieving, storing and supplying human tissue is described. In publishing the activities of the tissue bank at Peterborough, we wish to encourage others to consider the availability of human tissue in their locality. We recommend a strict legal and ethical code, particularly in relation to fully informed consent.
Toxicological Evaluation of Surface-water Samples in Sensitised Cultured Fish Cells Compared with the Microtox® Method
Authors: Dierickx Paul J. , Van der Wielen Claire , Lemaire Sandra
Published: April 2019

A previously described neutral red uptake-inhibition assay on cultured fathead minnow (FHM) fish cells revealed a good correlation with fish toxicity data. The assay was made more sensitive by reducing the number of cells, by using a longer treatment period, and by simultaneous treatment of the cells with sodium dodecyl sulphate and L-buthionine ( S,R)-sulphoximine. The fluorimetrically quantified protein content was then used as the endpoint. The results were compared with toxicity data obtained by the Microtox® method with the bacterium, Vibrio fischeri. A series of 82 surface-water samples were investigated. Cell quantity-dependent and sample concentration-dependent reductions in total protein content were observed. In all, 32 samples were toxic in the FHM assay. Of the seven samples that were shown to be toxic in the Microtox assay, only three were also toxic in FHM cells. No linear relationship was found for the toxicity results obtained with the two methods. Further research to explain the significance of the many positive responses and to identify possible confounding factors will increase the reliability of the sensitised FHM cell assay.
In Vitro Cytotoxicity of Four Different Buffers for Use in Peritoneal Dialysis
Authors: Järkelid Lena E. , Svensson Eva , Deppisch Reinhold , Wieslander Anders
Published: April 2019

Various buffers can be used in fluids for peritoneal dialysis (PD). Lactate is the most frequently used buffer, but bicarbonate and pyruvate have been suggested as more bio-compatible alternatives. In the past, acetate was used as a buffer in PD fluids, but was abandoned after being linked with sclerosing peritonitis and loss of ultrafiltration. When a new buffer for PD fluids is introduced, one of its most important characteristics is that it must be non-toxic, i.e. that it does not influence fundamental cellular functions. The aim of this study was to investigate the basal cytotoxicity of bicarbonate, acetate, lactate and pyruvate at neutral pH. As target cells, we used cultured mouse fibroblast-like L-929 cells, a well-known cell line approved by the authorities for regulatory use, and primary human mesothelial cells, which are the cells that line the peritoneal cavity and are exposed to the PD fluid in vivo. Pyruvate was more cytotoxic than lactate and bicarbonate, and no significant difference in cytotoxicity was found between lactate and bicarbonate. The human mesothelial cells were more sensitive to exposure to pyruvate than the L-929 fibroblast-like cells, but less sensitive to exposure to pure PD fluids. Thus, we recommend that both types of cell are used for the evaluation of the bio-compatibility of PD fluids.
Toxicity Testing of Polymer Materials for Dialysis Equipment: Reconsidering In Vivo Testing
Authors: Kjellstrand Per , Lindqvist Eva , Nilsson-Thorell Carin
Published: April 2019

The rationale for preclinical testing of plastic materials for medical devices is the protection of patients from leachable toxic substances. A controversial and costly part of this testing is the use of animal in vivo procedures. The objective of the present study was to analyse the importance of in vivo tests in relation to the decision to approve or not to approve materials for use. A total of 1044 plastic materials were analysed by employing chemical, in vitro and in vivo tests: 5708 in vivo tests were performed. In only one out of 2650 systemic injection tests on mice did a material fail. As that material also failed in chemical tests, the systemic injection test had no influence on the decision not to approve the material. Intradermal irritation (2644 tests), implantation (398 tests) and sensitivity (11 tests) procedures on rabbits and guinea-pigs were the other in vivo tests. However, in all except three cases, the same decision on whether or not to use a material would have been reached without any of these in vivo tests. Thus, little security appears to be gained from the in vivo tests, and abandoning them would save resources, probably without any additional risk.
Iron-induced Lipid Peroxidation in Human Skin-derived Cell Lines: Protection by a Red Orange Extract
Authors: Morini Flora , Dusatti Fabiola , Bonina Francesco P. , Saija Antonella , Ferro Margherita
Published: April 2019

Although photodamage and photoprotection have already been extensively studied in cultured cells, few data have been reported in the literature regarding the in vitro behaviour of skin cells toward a chemical stress, such as iron-induced lipid peroxidation. We investigated the susceptibilities of two human skin-derived cell lines (NCTC 2544 keratinocytes and HFFF2 fibroblasts) to lipid peroxidation induced by FeSO4/histidine, FeSO4/ascorbate and Fe2(SO4)3/ADP. NCTC 2544 cells were more susceptible than HFFF2 cells to lipid peroxidation (assessed by measuring the content of malondialdehyde [MDA]) with iron/ascorbate and iron/ADP as pro-oxidants whereas, with iron/histidine, the same level of MDA production was achieved (about 10nmol/mg protein) in the two cell populations. On the basis of these results, one experimental model (iron/histidine) was selected to assess the protective effect of a mixture of two classical antioxidants, Trolox C™ (50μM) and Vitamin C (1mM), added to the cell cultures according to various protocols. The maximal decrease of MDA production in both cell lines was obtained when the antioxidant mixture and the pro-oxidant were added simultaneously to the cultures. By using the same experimental design, NCTC 2544 and HFFF2 cells were exposed to a standardised extract of red oranges (ROE; 0.025–0.5mg/ml), the main active principles of which (anthocyanins 3.1%, hydroxycinnamic acid 2.07%, and flavanone glycosides 8.1%) possess antioxidant activity. Cells treated with ROE, that were still over 90% viable, as evaluated by means of neutral red uptake and tetrazolium salt reduction tests, showed a significant and dose-dependent inhibition of MDA production. This study provides new information about the behaviour of cultured skin cells exposed to chemically induced oxidative stress, and provides further support to the possibility of using skin-derived human cell lines in the evaluation of the effectiveness of antioxidant ingredients for new drugs and/or cosmetics.
Cytotoxicity of Cadmium, Selenium, Zinc and Copper to Mouse Myeloma Sp2/0 Cells as Measured by the MTT Assay
Authors: Remez Inessa , Rabkin Leonid , Veksler Hackel , Baumane Maija
Published: April 2019

As part of our general programme to test the toxicity in vitro of heavy metals and, in particular, the interactive cytotoxicity of combinations of these metals, the MTT (tetrazolium) assay was used to test the cytotoxicities of cadmium [Cd(O2CCH3)2], selenium (Na2SeO3), zinc (ZnSO4) and copper (CuSO4) with an established mouse myeloma cell line (Sp2/0). The influence on the cytotoxicity of cadmium of added subtoxic concentrations of the other three metals was also studied. The IC50 values (50% inhibitory concentrations) to the Sp2/0 cells were as follows: Cd — 10μM, Se — 1μM, Zn — 10μM and Cu — 100μM. When selenium, zinc and copper at high, subtoxic concentrations (10nM, 1μM and 1μM, respectively) were each combined with clearly toxic concentrations of cadmium (10μM and 100μM), the cytotoxicity increase was significant for the cadmium-zinc combination. Previous studies showed that cadmium-zinc combinations at low concentration (5 × 10–9M) had a synergistic stimulatory effect on cell proliferation in vitro. The present study showed that a toxic concentration of cadmium (0.1mM) and a sub-toxic concentration of zinc (1μM) have a synergistic cytotoxic effect in vitro.
Structure–permeability Relationships for Transcorneal Penetration
Authors: Worth Andrew P. , Cronin Mark T.D.
Published: April 2019

The main objective of this study was to determine, by using a literature data set of 112 corneal permeability measurements, whether a statistically significant structure–permeability relationship (SPR) for predicting the transport of structurally diverse chemicals across the rabbit cornea could be developed on the basis of just two physicochemical properties, the logarithm of the octanol–water partition coefficient (logP), and molecular weight (MW), which provide measures of lipophilicity and molecular size, respectively. A secondary objective was to identify additional physicochemical properties that are predictive of corneal permeability, and to use these as descriptors in an SPR. To achieve the first objective, multiple linear regression of corneal permeability against logP and MW was performed, and was found to produce a statistically significant (p < 0.001) SPR with stable coefficients. The second objective was achieved by carrying out stepwise regression analysis of corneal permeability against a number of calculated physicochemical properties; this identified three variables, representing lipophilicity (logP), molecular shape (the third-order kappa index [κ3]), and hydrogen-bonding capacity (nH). A three-variable SPR based on these descriptors was also statistically significant (p < 0.001) and had stable coefficients. Mechanistically based SPRs, such as the ones reported in this paper, could be used during the drug discovery process as a first step in the prediction of transcorneal delivery.
New Aspects of Cholecystokinin Processing and Visualisation in the Rat Brain by Using Antibodies Raised in Chickens and Rabbits
Authors: Schade Rüdiger , Henklein Peter , Harms Christoph , Jonas Ludwig , Lautenschlager Marion , Schöneberg Torsten , de Weerth Andreas , Hlinak Andreas , Hörtnagl Heide
Published: April 2019

Neuropeptides such as cholecystokinin (CCK) are subjected to a stepwise enzymic degradation (peptide processing) during axonal transport from the somata to the nerve terminals. This results in a continual change in the primary, secondary and tertiary structures of the peptide. Thus, an antibody raised against a selected sequence of the propeptide may recognise the antigen only at a certain stage of its “ontogenesis”. To address these difficulties, a set of antibodies with differing specificities and origins (chicken, rabbit) were used to visualise neuronal CCK by immunohistochemical methods in rat–brain sections (RBS) and in rat primary neuronal cultures (PNC). The specificity of the antibodies was analysed by using a dot-blot assay and a radioimmunoassay. Marked differences in the reactivities of the various antibodies were observed and related to the antigen used, inter-individual variations and the animal species. In the cortex, various types of CCK-immunoreactive neurons were found, including spindle-shaped or button-shaped neurons and pseudo-unipolar neurons. However, in contrast to mammalian antibodies, several of the chicken antibodies recognised cortical pyramidal neurons in both RBS and PNC without pretreatment with colchicine. Evidence has been obtained in both RBS and PNC that an antibody with a defined specificity may not visualise the entire neuron, but only distinct parts of it, possibly depending on the actual molecular structure of the neuropeptide present at a specific locus of the neuron. A complete mapping of a neuronal peptide that is processed during axonal transport can only be achieved by using a set of different antibody-specificities.
The Humoral Immune Response and the Productivity of Laying Hens Kept on the Ground or in Cages
Authors: Erhard Michael H. , Özpinar Haydar , Bilal Tanay , Abbas Ysmail , Kutay Can , Eseceli Hüseyin , Stangassinger Manfred
Published: April 2019

The effects of two different keeping systems on the humoral immune response and productivity were compared for 80 laying hens, divided into four groups. Two groups, each of 20 hens, were kept on the ground and two were kept in cages. All the birds were immunised subcutaneously with human serum immunoglobulin G at a dose of 100μg per injection. The immunisations were performed twice at 4-week intervals. The lipopeptide Pam3Cys-Ser-(Lys)4 was used as an adjuvant at a dose of 0.25mg per injection in one group from each housing system. In the second group from each housing system, the hens were immunised without any adjuvant (antigen control groups). The mean egg yield was significantly higher in both the antigen control group and the adjuvant group, when laying hens were kept in cages. Total egg weight remained constant in both of the housing systems. Keeping hens in cages resulted in higher mean specific antibody titres and mean immunoglobulin Y concentrations in the egg yolk.
An Evaluation of Mechanistic In Vitro Tests for the Discrimination of Photoallergic and Photoirritant Potential
Authors: Lovell William W. , Jones Penny A.
Published: April 2019

Photochemical tests were used to discriminate between photoallergens and photoirritants. UV absorption spectrometry was employed to identify chemicals that absorbed sunlight wavelengths and which required further testing. Photoallergic potential was assessed by studying photobinding of the test chemicals to human serum albumin. Photobinding was determined by increased UV absorbance of the protein fraction after gel filtration chromatography. Photooxidation of histidine was used to screen for a mechanism of photoirritancy. Efficient photooxidisers can be considered to be photoirritant rather than photoallergic. The substances selected for the EU/COLIPA phototoxicity project were tested. There were 14 photoirritants (three tested as both free acid/base and salts, i.e. a total of 17 samples), four photoallergens, three of which were photoirritant and photoallergenic (i.e. 17 photoirritants and seven photoallergens) and six “negatives” (four clearly non-phototoxic and two unclear). UV spectrometry showed that 28 of the 30 substances absorbed sunlight significantly and had the potential for adverse photoreaction. Six of seven photoallergens were identified as such by the photo-binding assay. Most photoirritants also caused photomodification of protein, but eleven of these photooxidised histidine efficiently and so were classified as photoirritants. Four photoirritants remained falsely predicted as photoallergens. Two photoirritants were negative for both photo-modification of protein and histidine photooxidation. Four chemicals negative in vivo were negative in vitro. The remaining two chemicals could not be classified, because of unclear data both in vivo and in vitro. Therefore, the in vitro test battery was useful for the discrimination of photoallergic and photoirritant potential.
In Vitro Preclinical Lead Optimisation Technologies (PLOTs) in Pharmaceutical Development
Authors: Atterwill Christopher K. , Wing Mark G.
Published: April 2019

The explosion of genuine high throughput technologies has allowed large compound libraries to be screened with ever-increasing biological specificity, exacerbating the problem of lead candidate selection for subsequent drug development. To avoid creating a bottleneck, compounds identified from the high throughput screens undergo lead optimisation by employing medium-throughput screens which permit ranking in terms of their basic absorption, distribution, metabolism, excretion (ADME) and toxicological properties. The historical role of the preclinical scientist in the drug discovery/development continuum has been to perform ADME and toxicology studies, simply to support the regulatory submission of lead candidates. This situation is, however, changing with the development of preclinical lead optimisation technologies facilitating the selection of leading candidates, thereby bridging the gap between high throughput efficacy screens and conventional safety assessment programmes.
Scientific Progress and Animal Welfare: Conflicting Goals or Complementary Opportunities?
Authors: Rothwell Nancy
Published: April 2019

In view of the recent controversy regarding the perceived need by scientists for animal experiments on the one hand, and the concerns expressed by animal-welfare groups on the other, ATLA has commissioned this guest editorial by Professor Nancy Rothwell, Professor of Physiology, University of Manchester, and Chairman of the UK Life Sciences Animal Science Group.
Molecular Profiling and Prediction of Toxicity
Authors: Schuppe-Koistinen Ina
Published: April 2019

The rapid progress in genomics and proteomics technologies creates a unique opportunity for understanding the mechanisms of action and the toxicities of compounds. The application of gene-expression and protein-expression profiling promises to improve the predictive power of safety assessment, resulting in the development of drug candidates with a higher efficacy and a lower toxicity. Additionally, in the later phases of drug development, surrogate markers of treatment efficacy and toxicity can be applied to optimise the monitoring of pre-clinical and clinical studies.
Biochemical Responses in Aquatic Plants as Markers of Environmental Contamination
Authors: Lindström-Seppä Pirjo , Urban Katalin , Honkalampi-Hämäläinen Ulla , Roy Sashwati
Published: April 2019

This short review gives several examples of the current status of xenobiotic bio-transformation reactions and oxidative stress responses in plants as biomarkers of organic pollution in aquatic environments. Based on previous basic knowledge, several biomonitoring programmes have been successfully applied during the last decade.
Caspase-3 Activity and Carbonyl Cyanide m-Chlorophenylhydrazone-induced Apoptosis in HL-60 cells
Authors: Mlejnek Petr
Published: April 2019

The role of caspase proteases in carbonyl cyanide m-chlorophenylhydrazone (CCCP)-induced apoptosis of human promyelocytic HL-60 cells was examined. Treatment of HL-60 cells with micromolar concentrations of CCCP resulted in cell death, with typical apoptotic features such as chromatin condensation, formation of apoptotic bodies, nucleosomal fragmentation of DNA and a distinct increase in caspase-3 activity. The results, however, indicated that full caspase-3 inhibition by the selective inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethyl ketone (Z-DEVD-FMK) did not prevent cell death, nor did it affect the manifestation of apoptotic hallmarks, including apoptotic bodies formation and nucleosomal DNA fragmentation. The only distinct effect that Z-DEVD-FMK exhibited was to retard the disruption of the plasma membrane. We therefore assume that caspase-3 activity itself is not essential for the manifestation of apoptotic features mentioned above. Similarly, the pan-specific caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD-FMK) did not prevent cell death. On the contrary, Z-VAD-FMK completely prevented DNA cleavage and apoptotic body formation, but it failed to completely counteract chromatin condensation. Thus, in the presence of Z-VAD-FMK, application of CCCP concentrations that otherwise induced apoptosis, resulted in the appearance of two morphologically different groups of dead cells with intact DNA. The first group included cells with necrotic-like nuclear morphology, and therefore could be taken as being “truly” necrotic in nature, because they had intact DNA. The cells of the second group formed small single-spherical nuclei with condensed chromatin. In spite of having intact DNA, they could not be taken as “truly” necrotic cells. It is evident that in the experimental system, caspase proteases play an essential role in the formation of apoptotic bodies and in the cleavage of nucleosomal DNA, but not in the condensation of chromatin. Therefore, it is likely that the choice between cell death modalities is not solely a matter of the caspase proteases present.
Pyruvate-induced Long-term Maintenance of Glutathione S-Transferase in Rat Hepatocyte Cultures
Authors: Vanhaecke Tamara , Foriers André , Geerts Albert , Shephard Elizabeth A. , Vercruysse Antoine , Rogiers Vera
Published: April 2019

The addition of pyruvate to the culture medium has been reported to improve the maintenance of P450-dependent enzyme expression in primary rat hepatocyte cultures. In this study, the effects of 30mM pyruvate on cell morphology, albumin secretion and glutathione S-transferase (GST) expression were investigated as a function of the time in culture. The effect of triiodothyronine (T3) exposure on GST expression was also measured in pyruvate-treated cultures. Transmission electron microscopy showed that untreated hepatocytes deteriorated after culture for 7 days, whereas the morphology of the pyruvate-treated cells was similar to that observed in intact liver tissue. The albumin secretion rate was significantly higher in rat hepatocytes exposed to pyruvate than in control cells. In the presence of pyruvate, μ and α class GST activities were well maintained, whereas GST π activity was increased over the entire culture period. HPLC analysis revealed that the complement of GST subunits present in hepatocytes is altered during culture with pyruvate: μ class proteins remained relatively constant, whereas a decrease in the a class content was accompanied by a strong increase in GST subunit P1 (GSTP1). The induction of GSTP1 was confirmed at the mRNA level. In control cultures, π class GST activity was increased, but total, μ, and α class GST activities continuously declined as a function of culture time and became undetectable beyond 7 days in culture. At the protein and mRNA levels, a much smaller increase in GSTP1 was observed than in the pyruvate cultures. When the pyruvate-treated cell cultures were exposed to T3, an inhibitory effect on GST activities and proteins was found. These results indicate that this simple culture model could be useful for studying the expression and regulation of GST.
In Vitro Assessment of Cytotoxicity and Carcinogenic Potential of Chemicals: Evaluation of the Cytotoxicity Induced by 58 Metal Compounds in the Balb/3T3 Cell Line
Authors: Mazzotti Francesca , Sabbioni Enrico , Ghiani Michela , Cocco Barbara , Ceccatelli Raffaella , Fortaner Salvador
Published: April 2019

A new, mechanistically based, in vitro strategy involving Balb/c 3T3 clone A31-1-1 mouse embryo fibroblasts has been proposed for the determination of the carcinogenic potential of inorganic chemicals, in order to establish priority of metal compounds to be tested and, whenever possible, to compare the in vitro results with the corresponding in vivo data. As a first step in this research, this study reports on the cytotoxic effects of 58 metal compounds in the Balb/3T3 cell line. After harmonisation and standardisation of the Balb/3T3 protocol, cells were exposed for 72 hours to a fixed dose (100μM) of 58 individual compounds. The cytotoxicity induced by some metal compounds was found to be related to their chemical form (for example, Cr(NO3)3 and Na2CrO4), suggesting that the Balb/3T3 cell line is a valuable cellular model in relation to this aspect of metal speciation. The results of the systematic study on the metal-induced cytotoxic effects in the Balb/3T3 cell line could be arbitrarily classified into three groups according to the degree of cytotoxicity. Group I includes 26 species that induced no observable effect or only a slight cytotoxic effect; Group II includes 13 metal compounds that exhibited an obvious degree of cytotoxicity; and Group III includes 19 metal species that displayed a strong cytotoxic response. Metal compounds of Groups II and III are considered to be of the highest priority for setting of dose–effect relationships for a subsequent in vitro study on metal-induced concurrent cytotoxicity and morphological transformation in the Balb/3T3 cell line.
The Role of ECVAM in Promoting the Regulatory Acceptance of Alternative Methods in the European Union
Authors: Worth Andrew P. , Balls Michael
Published: April 2019

The roles played by the European Centre for the Validation of Alternative Methods (ECVAM) and its advisory committee, the ECVAM Scientific Advisory Committee (ESAC), in the evolution of alternative methods are described. Particular emphasis is given to the process by which ECVAM and the ESAC assess the scientific validities of alternative methods, and, in appropriate cases, initiate the progression of scientifically validated methods toward regulatory acceptance.
Human Haemoclassification by use of Specific Yolk Antibodies Obtained after Immunisation of Chickens against Human Blood Group Antigens
Authors: Calzado Esteban Gutiérrez , Garrido Rosa Mireya García , Schade Rüdiger
Published: April 2019

Polyclonal antibodies, widely used in research and diagnostics, are conventionally isolated from the blood of immunised mammals, especially rabbits. The fact that antibodies can also be detected in the yolk of eggs laid by immunised hens, led to the development of yolk antibody technology (IgY-technology) as an alternative method that is less stressful to animals. This technology has become a worthwhile alternative to the blood-dependent techniques. Furthermore, because of the phylogenetic distance between birds and mammals, avian antigens have a very specific immune response to highly conserved antigens of mammals, such as human erythrocyte antigens. To evaluate the humoral immune response of hens immunised with human red erythrocyte antigens, 22 White Leghorn hens were kept in cages and immunised with total red blood cells or stroma of the human rhesus positive (Rh+) system (D antigen) by weekly intramuscular and intravenous injections, without the use of an adjuvant. The haemag-glutination assay was used to evaluate the dynamics of the production of IgY antibodies against human erythrocyte antigens, and single radial immunodiffusion was used to evaluate the amount of total IgY in de-lipidated supernatants from egg yolk. The highest titres were observed four weeks after the first immunisation, and these remained stable for up to seven weeks for the intravenous route. Positive reactivity against human erythrocyte antigens A, B and O was demonstrated in de-lipidated supernatants from the egg yolks of immunised hens. The strongest reaction was observed against blood group O Rh+ (O+). Furthermore, the antibodies obtained recognised erythrocyte antigens that belong to neither the Rh system nor the Landsteiner-Weiner antigen. This finding opens the possibility of using hens instead of rabbits to produce polyclonal antibodies for human haemoclassification.