Reproducible preparation of spheroids of pancreatic hormone positive cells from human iPS cells: An in vitro study

BackgroundTransplantation of islets of Langerhans is regarded as a promising therapy for type 1 diabetes. A large number of β-cells are required for the treatment of human type 1 diabetes. Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, have been considered as new sources for cell replacement therapy.MethodsCell aggregates were prepared from human iPS cells using agarose microwell plates and differentiated into pancreatic endocrine cells by changing the culture media with different additives.ResultsAfter 20 days of culture, approximately 30% of cells in aggregates were positive for C-peptide. After another 14 days in culture, the cells gained an ability to alter C-peptide release in response to changes in the glucose concentration.ConclusionsUniform aggregates of human iPSCs were easily prepared on agarose microwell plates and efficiently differentiated into the pancreatic endocrine lineage. Thus, aggregate culture is a suitable method for preparing islet-like aggregates from human iPSCs.General significanceOur results indicate that the microwell plate is suitable for scaling up the preparation of pancreatic endocrine cells from human iPS cells in a robotic system.     

Microsporum canis en gatos dermatológicamente sanos en Temuco,Chile

BackgroundMicrosporum canis is the most common cause of feline dermatophytosis and the most pathogenic fungus isolated from the skin and hair of healthy cats. Cats are considered to be the natural reservoir and infection sourse of this disease in human and domestic animals.AimsKnowing the M. canis frequency in the dermatological healthy cat population of Temuco city, Chile.MethodsFifty cat samples were collected irrespective sex or race. Cats' ages were between 2 months and 12 years old, and the animals were treated at the Veterinary Clinical Hospital of the Universidad Católica de Temuco, or in three private clinics from this city. Tissue and hair samples were collected using two sampling techniques: hair extracting tweezers and the Mariat & Tapia method. For the clinical diagnosis, the Wood's lamp was used. Hairs were microscopically observed followed by a culture using Sabouraud agar and Lactrimel agar. M.canis was isolated in 30 cats (60%).ResultsThere were no statistically significative differences when parameters such as age, sex and race were taking into account. Differences between the use of Sabouraud agar and Lactrimel agar were not registered. It was determined that the Mariat & Tapia method was able to detect more dermatophytes than the collecting tweezers method. These differences were statistically significative.ConclusionsThe percentage of M. canis isolation obtained in this work remarks the role of healthy cats in the transmission of these dermatophytes to humans and other animals.     

Animals,Archetypes, and Popular Culture: Tales from the Tabloid Press

Abstract

This paper characterizes the portrayal of animals and human-animal relations in one genre of American popular culture—the “supermarket” tabloid press. A total of 789 animal-related stories and photographs in 82 issues of four tabloid magazines were analyzed according to theme. The items fell into nine categories in which animals were portrayed as objects of affection, saviors, threats, victims, things to be used, sex objects, imaginary and mythological beings, surrogate humans, and objects of wonder. It is argued that these themes represent archetypes reflecting the roles that animals have had in human cultural and psychological life since the historical origins of our species.     

Rapid culture of human keratinocytes in an autologous,feeder-free system with a novel growth medium

Background aimsThe ability to culture human keratinocytes is beneficial in the treatment of skin injury and disease, as well as for testing chemicals in vitro as a substitute for animal testing.ResultsWe have identified a novel culture medium for the rapid growth of keratinocytes from human skin. “Kelch's medium” supports keratinocyte growth that is as rapid as in the classical Rheinwald and Green method, but without the need for cholera toxin or xenogeneic feeder cells. It enables keratinocytes to out-compete co-cultured autologous fibroblasts so that separation of the epidermis from the dermis is no longer required before keratinocyte culture. Enzymatic digests of whole human skin can therefore be used to generate parallel cultures of autologous keratinocytes, fibroblasts and melanocytes simply by using different cell culture media.ConclusionsThis new keratinocyte medium and the simplified manufacturing procedures it enables are likely to be beneficial in skin engineering, especially for clinical applications.     

某三甲医院儿童和成年女性尿培养病原菌分布和耐药性分析

摘要 目的：了解尿路感染儿童和成年女性尿培养病原菌种类和耐药性的差异,为临床合理选用抗菌药物提供依据。方法：分别收集2018年1月~2019年12月期间在我院住院的尿路感染儿童的尿培养标本1618份和尿路感染成年女性的尿培养标本1044份,分析其病原菌的分布和耐药性。结果：1618份儿童尿培养标本中分离出267株病原菌,居首位的病原菌是屎肠球菌,占43.82%（117/267）;1044份成年女性尿培养标本中分离出139株病原菌,居首位的病原菌是粪肠球菌,占28.78%（40/139）。在两种人群尿培养病原菌中大肠埃希菌和肺炎克雷伯菌构成比均分别为第二位和第三位。儿童尿培养屎肠球菌对青霉素G、氨苄西林、环丙沙星等喹诺酮类药物的耐药率高于成年女性尿培养粪肠球菌,对克林霉素、奎奴普丁/达福普汀、四环素的耐药率低于成年女性尿培养粪肠球菌（P＜0.05）;未发现对高浓度庆大霉素、高浓度链霉素、利奈唑胺、万古霉素、替加环素耐药的肠球菌。儿童尿培养大肠埃希菌对氨苄西林/舒巴坦、头孢吡肟的耐药率均高于成年女性尿培养大肠埃希菌（P＜0.05）。儿童尿培养肺炎克雷伯菌对头孢哌酮/舒巴坦、氨苄西林/舒巴坦、哌拉西林/他唑巴坦、氨曲南、厄他培南、亚胺培南、美洛培南、呋喃妥因、头孢唑啉等头孢菌素类药物的耐药率高于成年女性尿培养肺炎克雷伯菌（P＜0.05）。结论：尿路感染儿童和成年女性尿培养病原菌均以肠球菌为主,大肠埃希菌和肺炎克雷伯菌构成比分别为第二位和第三位,两种人群尿培养主要病原菌耐药性均有不同程度的差异,临床医生应根据尿培养和药敏结果合理用药。     

Ambiguous Mice,Speaking Rats: Crossing and Affirming the Great Divides in Scientific Practice

ABSTRACT

During 2009–10, I conducted ethnographic fieldwork with 31 immunologists, virologists, and neuroscientists working with either rats or mice. I encountered how the conceptual and physical bounds that have traditionally separated nature from culture, specie from specie, human from animal, are crossed, blurred, and reasserted. In this ambiguous zone, a scientific incuriosity about animals themselves persists, in the practice of inquiring into animal bodies and minds to produce insights into human health and its betterment. This privileging of human health bypasses animals themselves in favor of a view of them as human similars and prone objects, wholly available to persons, and affirms the Heideggarian thesis, that science occupies an arrogated position in modernity. Such incurious encounters with animals produced ideas and pronouncements about the close biological and genetic similarities that humans and animals share, that scientists in my study called “biokinship” and “genekinship.” These terms indicate both a close relation between animals and persons, but they also present the terms upon which hierarchical relations between humans and animals might be arrayed. Equally present among the scientists with whom I worked was a curiosity about animals themselves. This manifested in understandings and articulations of animals as beings with whom one might make a relationship in which mutually understood communication was possible. Attendant to this curiosity about animals themselves was an awareness scientists in my study had of what these relationships, or what I have called fleshy kinships with rats and mice, might mean for scientific practice, for good science, and for human–animal relatedness in the laboratory. This ambiguous situation calls for analytic attention to biotic materiality and process, but equally for attention to rodents as beings with whom scientists interact on an everyday basis, and with whom they form communicative relations.     

Zebrafish Tg(hb9:MTS-Kaede): a new in vivo tool for studying the axonal movement of mitochondria

ObjectivesDeregulation of axonal transport in neurons is emerging as the major cause of many neurodegenerative diseases in human, such as Charcot-Marie-Tooth (CMT) neuropathy. However, little is known about how mitochondria move in vivo and whether cell culture systems truly represent what happens in living animals. Here we describe the generation of a new zebrafish transgenic line that specifically allows to study mitochondrial dynamics in motor neurons and its application to analyse mitochondrial movement in zebrafish models expressing CMT2A causing mutations.MethodsThe Tol2 transposon system was used to generate a transgenic zebrafish line expressing the photoconvertible fluorescent protein Kaede in mitochondria of motor neurons. Mitochondrial shape and movement were monitored by time-lapse confocal live imaging and measured by kymograph analysis. The effects of two well-known CMT causing mutations, L76P and R94Q substitutions in MFN2, were then investigated with the same methods.ResultsWe generated the transgenic zebrafish Tg(hb9:MTS-Kaede) line with genetically labelled mitochondria in motor neurons. Kaede protein was correctly and stably targeted to mitochondrial matrix while retaining its photoconvertibility, thus qualifying this model for in vivo studies. Expression of the L76P and R94Q mutations reduced mitochondrial movement in axons and altered mitochondrial distribution in distinct ways.Conclusions and general significanceThese findings confirm previously published data obtained in cell cultures and strengthen the hypothesis of different mechanism of action of the two MFN2 mutations. Considering the number of neurodegenerative diseases associated to mitochondrial dynamics, the Tg(hb9:MTS-Kaede) zebrafish line is a promising model to study in vivo alterations of mitochondrial transport underlying human diseases.     

Dogmatisms and Catechisms — Ethics and Companion Animals

Abstract

The current reconsideration of the place in nature of human beings unfortunately continues to be an acrimonious one. All too often the debate is more akin to a warlike encounter where each side attempts to gain control or the upper hand than a search for points of agreement. Given this context, it is important to entertain views that emanate from different cultural traditions as a way to infuse the debate with new life. Students of Native American culture have consistently pointed out that the essential concepts of life balance and reciprocity represented there may serve as useful points of consideration as we struggle with the appropriate relationships with animals and nature. This article presents a representative Zuni story, told by Governor Robert E. Lewis, that illustrates these notions.     

Sero-epidemiological survey of Coxiella burnetii in livestock and humans in Tana River and Garissa counties in Kenya

BackgroundCoxiella burnetii is a widely distributed pathogen, but data on its epidemiology in livestock, and human populations remain scanty, especially in developing countries such as Kenya. We used the One Health approach to estimate the seroprevalance of C. burnetii in cattle, sheep, goats and human populations in Tana River county, and in humans in Garissa county, Kenya. We also identified potential determinants of exposure among these hosts.MethodsData were collected through a cross-sectional study. Serum samples were taken from 2,727 animals (466 cattle, 1,333 goats, and 928 sheep) and 974 humans and screened for Phase I/II IgG antibodies against C. burnetii using enzyme-linked immunosorbent assay (ELISA). Data on potential factors associated with animal and human exposure were collected using a structured questionnaire. Multivariable analyses were performed with households as a random effect to adjust for the within-household correlation of C. burnetii exposure among animals and humans, respectively.ResultsThe overall apparent seroprevalence estimates of C. burnetii in livestock and humans were 12.80% (95% confidence interval [CI]: 11.57–14.11) and 24.44% (95% CI: 21.77–27.26), respectively. In livestock, the seroprevalence differed significantly by species (p < 0.01). The highest seroprevalence estimates were observed in goats (15.22%, 95% CI: 13.34-17.27) and sheep (14.22%, 95% CI: 12.04–16.64) while cattle (3.00%, 95% CI: 1.65–4.99) had the lowest seroprevalence. Herd-level seropositivity of C. burnetii in livestock was not positively associated with human exposure. Multivariable results showed that female animals had higher odds of seropositivity for C. burnetii than males, while for animal age groups, adult animals had higher odds of seropositivity than calves, kids or lambs. For livestock species, both sheep and goats had significantly higher odds of seropositivity than cattle. In human populations, men had a significantly higher odds of testing positive for C. burnetii than women.ConclusionsThis study provides evidence of livestock and human exposure to C. burnetii which could have serious economic implications on livestock production and impact on human health. These results also highlight the need to establish active surveillance in the study area to reduce the disease burden associated with this pathogen.     

脂肪干细胞的诱导分化及其来源的研究

目的:通过组织块培养法得到脂肪干细胞(adipose-derived stem cells,ADSCs),探讨其诱导分化潜能,并初步研究ADSCs的来源。方法:用脂肪组织块培养法培养原代人ADSCs。第三代ADSCs进行成脂和成骨诱导分化,分别用油红O和茜素红S染色进行鉴定。脂肪组织块培养七天后取脂肪组织进行Hematoxylin-eosin Staining(HE)染色观察ADSCs组织分布。结果:用脂肪组织块培养法成功培养出原代人ADSCs。ADSCs传代到第8代,依然保持着良好的增殖能力和细胞形态。ADSCs能成功诱导成脂肪细胞和骨细胞。通过对培养七天后的脂肪组织块进行HE染色,发现ADSCs主要分布在脂肪组织的间质血管和结缔组织周围。结论:用脂肪组织块培养出来的ADSCs具有成脂和成骨分化的潜能。ADSCs主要定位于间质血管和结缔组织周围。     

Successful Treatment of Severe Tungiasis in Pigs Using a Topical Aerosol Containing Chlorfenvinphos,Dichlorphos and Gentian Violet

BackgroundIn endemic communities, zoonotic tungiasis, a severe skin disease caused by penetrating female sand fleas, is a public health hazard causing significant human and animal morbidity. No validated drugs are currently available for treatment of animal tungiasis. Due to the reservoir in domestic animals, integrated management of human and animal tungiasis is required to avert its negative effects.ConclusionsThe study demonstrates that a topical treatment based on chlorfenvinphos, dichlorphos and gentian violet is highly effective against pig tungiasis. Due to its simplicity, the new approach can be used for the treatment of individual animals as well as in mass campaigns.     

Human platelet lysate is a feasible candidate to replace fetal calf serum as medium supplement for blood vascular and lymphatic endothelial cells

Background aimsAs angiogenic and lymphangiogenic key players, endothelial cells (ECs) are promising candidates for vascular regenerative therapies. To culture ECs in vitro, fetal calf serum (FCS) is most often used. However, some critical aspects of FCS usage, such as possible internalization of xenogeneic proteins and prions, must be considered. Therefore, the aim of this project was to determine if human platelet lysate (hPL) is a suitable alternative to FCS as medium supplement for the culture of blood vascular and lymphatic endothelial cells.MethodsThe usability of hPL was tested by analysis of endothelial surface marker expression, metabolic activity and vasculogenic potential of outgrowth ECs (OECs), human umbilical vein ECs (HUVECs), and lymphatic ECs (LECs).ResultsExpression of EC markers CD31, VEGFR2, VE-cadherin and CD146 did not differ significantly between the EC types cultured in FCS or hPL. In addition, OECs, HUVECs and LECs formed tube-like structures on Matrigel when cultured in hPL and FCS. With the use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid assays, we found that the metabolic activity of OECs and LECs was slightly decreased when hPL was used. However, HUVECs and LECs did not show a significant decrease in metabolic activity, and HUVECs showed a slightly higher activity at low seeding densities.ConclusionsThe use of hPL on different EC types did not reveal any substantial negative effects on EC behavior. Thus, hPL appears to be a favorable candidate to replace FCS as a medium supplement in the culture of ECs.     

Understanding Nazi Animal Protection and the Holocaust

Abstract

It is well known that the Nazis treated human beings with extreme cruelty but it less widely recognized that the Nazis also took some pains to develop and pass extensive animal protection laws. How could the Nazis have professed such concern for animals while treating humans so badly? It would be easy to dismiss Nazi proclamations on animals as mere hypocrisy but there may be other explanations for the contradiction. For example, anecdotal reports and psychological evaluations of many prominent Nazis suggest they felt affection for animals but dislike of humans. Second, animal protection measures, whether sincere or not, may have been a legal veil to attack Jews and others considered undesirable. Third, the Nazis blurred moral distinctions between animals and people and tended to treat members of even the Master Race as animals at times. This article argues that at the core of the Nazi treatment of humans and animals was a reconstitution of society's boundaries and margins. All human cultures seek to protect what is perceived to be pure from that which is seen to be dangerous and polluting and most societies establish fairly clear boundaries between people and animals. In Nazi Germany, however, human identity was not contaminated by including certain animal traits but certain peoples were considered to be a very real danger to Aryan purity.     

Empathy and attitudes to animals

Abstract

There is increasing support for the idea that human attitudes to animals may be indicative of human–human empathy. This has implications for the treatment of empathy deficits and related anti-social behaviors. The purpose of the present study was to explicitly investigate links between human–human empathy and attitudes to animals. The Interpersonal Reactivity Index (IRI) and Animal Attitude Scale (AAS) were administered to 194 undergraduate Sociology and Psychology students. A significant correlation between empathy levels, gender, companion animal ownership and attitudes to animals was found. Implications of these findings are discussed.     

Three-dimensional organoid culture unveils resistance to clinical therapies in adult and pediatric glioblastoma

BackgroundGlioblastoma (GBM) is the most common primary brain tumor with a dismal prognosis. The inherent cellular diversity and interactions within tumor microenvironments represent significant challenges to effective treatment. Traditional culture methods such as adherent or sphere cultures may mask such complexities whereas three-dimensional (3D) organoid culture systems derived from patient cancer stem cells (CSCs) can preserve cellular complexity and microenvironments. The objective of this study was to determine if GBM organoids may offer a platform, complimentary to traditional sphere culture methods, to recapitulate patterns of clinical drug resistance arising from 3D growth.MethodsAdult and pediatric surgical specimens were collected and established as organoids. We created organoid microarrays and visualized bulk and spatial differences in cell proliferation using immunohistochemistry (IHC) staining, and cell cycle analysis by flow cytometry paired with 3D regional labeling. We tested the response of CSCs grown in each culture method to temozolomide, ibrutinib, lomustine, ruxolitinib, and radiotherapy.ResultsGBM organoids showed diverse and spatially distinct proliferative cell niches and include heterogeneous populations of CSCs/non-CSCs (marked by SOX2) and cycling/senescent cells. Organoid cultures display a comparatively blunted response to current standard-of-care therapy (combination temozolomide and radiotherapy) that reflects what is seen in practice. Treatment of organoids with clinically relevant drugs showed general therapeutic resistance with drug- and patient-specific antiproliferative, apoptotic, and senescent effects, differing from those of matched sphere cultures.ConclusionsTherapeutic resistance in organoids appears to be driven by altered biological mechanisms rather than physical limitations of therapeutic access. GBM organoids may therefore offer a key technological approach to discover and understand resistance mechanisms of human cancer cells.     

The Quantum Co-Evolution Unit: An Example of ‘Awa (Kava—<Emphasis Type="Italic">Piper methysticum</Emphasis> G. Foster) in Hawaiian Culture

The Quantum Co-Evolution Unit: An Example of ‘Awa (Kava— Piper methysticum G. Foster) in Hawaiian Culture. The process of co-evolution occurs in many kinds of relationships and on various scales. One example of a co-evolutionary relationship is that of a plant and a culture with which it interacts. Such relationships are dynamic and ever changing. Researchers have discussed this concept and its implications for decades, yet no quantifiable unit or standardized scale has been accepted with which to measure this change. The theoretical “quantum co-evolution unit” (QCU) is proposed as the smallest measurable scale of interactions between plants and people. A collection of QCUs for a linked plant and human population would be its “ethnobotanical population.” This could be measured at various points in time to quantify the changing relationships between plants and people. These models set up a structure to discuss methodologies for quantifying co-evolutionary relationships such as are seen in the evolution of ethnobotanical populations. The co-evolving relationship between ‘awa (kava—Piper methysticum) and Hawaiian culture is used as an example to illustrate this idea.     

Genetic stability of bone marrow-derived human mesenchymal stromal cells in the Quantum System

Background aimsThe Quantum® Cell Expansion System (Quantum; Terumo BCT, Inc, Lakewood, CO, USA) is a novel hollow fiber-based device that automates and closes the cell culture process, reducing labor intensive tasks such as manual cell culture feeding and harvesting. The manual cell selection and expansion processes for the production of clinical-scale quantities of bone marrow-derived human mesenchymal stromal cells (BM-hMSCs) have been successfully translated onto the Quantum platform previously. The formerly static, manual, in vitro process performed primarily on tissue culture polystyrene substrates may raise the question of whether BM-hMSCs cultured on a hollow fiber platform yields comparable cell quality.MethodsA rigorous battery of assays was used to determine the genetic stability of BM-hMSCs selected and produced with the Quantum. In this study, genetic stability was determined by assessing spectral karyotype, micronucleus formation and tumorigenicity to resolve chromosomal aberrations in the stem cell population. Cell phenotype, adherent growth kinetics and tri-lineage differentiation were also evaluated. HMSC bone marrow aspirates, obtained from three approved donors, were expanded in parallel using T225 culture flasks and the Quantum.ResultsBM-hMSCs harvested from the Quantum demonstrated immunophenotype, morphology and tri-lineage differentiation capacity characteristics consistent with the International Society of Cell Therapy standard for hMSCs. Cell populations showed no malignant neoplastic formation in athymic mice 60 days post-transplant, no clonal chromosomal aberrations were observed and no DNA damage was found as measured by micronucleus formation.ConclusionsQuantum-produced BM-hMSCs are of comparable quality and demonstrate analogous genetic stability to BM-hMSCs cultured on tissue culture polystyrene substrates.