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要旨 目的：管理栄養士・栄養士（以下，RD 等）の職場は，対人・フードサービスが含まれ，COVID-19 発生リスクが高い環境下である可能性が高い。本研究では，コロナ禍におけるRD 等の職場の現状を明らかにすることを目的とした調査研究結果を報告する。方法：2021 年10 ～ 12 月に典型的な職場で働くRD 等が選択され，コロナ対策に関する質問を含む自記式質問票に回答した。結果：72 人の回答者のうち，70% 以上が「不安」，65% が「情報不足」，51% が「人員不足」を感じていた。給食施設4 分野間では，特に高齢者施設では人員不足を感じている者が多く，有意差がみられた。コロナ対応情報は，医療90％，高齢者施設80％の参加者から得られ，コロナ禍における食事観察・コミュニケーションの機会の減少が報告された。さらに，回答者の約40％は，COVID-19 患者がいる職場での具体例を知りたいことがわかった。結論：緊急時に，少数人員で働くRD 等のプレッシャーを軽減できるサポートシステムを構築することが望ましい。, Objectives: Registered Dietitians’(RD) and Dietitiansʼ workplace involve interpersonal and food services for a wide range of people. Such workplaces can be high-risk environments for COVID-19 outbreaks. Herein, we report the results of a case study conducted to clarify the current state of the RD/Dietitians’ workplace during the COVID-19 pandemic. Methods: Participants working at typical workplaces between October and December 2021 were selected. They responded to a self-administered questionnaire, which included questions on workplace measures adopted during the COVID-19 pandemic. Results: More than 70% of the 72 respondents felt some anxiety about COVID-19 infection in their workplace, 65% felt a lack of information, and 51% felt a lack of personnel. Especially in the field of welfare for the elderly, many RD feel that there is a shortage of personnel, and there is a significant difference among the four fields that have food service facilities. The information on the extent of measures during the COVID-19 pandemic was obtained from participants in the medical (90%) and the welfare fields (80%). We found that during the COVID-19 pandemic, the opportunities for meal observation in patients and communication, decreased. In addition, approximately 40% of the respondents answered that they would like further details on the specific measures adopted at the workplace with COVID-19 patients. Conclusion: It is desirable to build a support system that can reduce the pressure on RD/Dietitians working in a place with few employees or one person in case of an emergency.

Background/Aim: Prostate cancer (PCa) is one of the most common types of cancer in men. Prostate-specific antigen (PSA) is currently the only biomarker used to screen for the risk of developing PCa. Because PSA tests may show false positives, identifying novel PCa-specific biomarkers would improve prediction and diagnosis at an early stage. Previously, we identified a number of genes/microRNAs (miRNAs) in prostate tissue as potential biomarkers of chronic prostatitis in a rat model of chemical-induced prostatitis. The current study aimed to evaluate their potential for use as translational, diagnostic markers in humans. Materials and Methods: We performed quantitative polymerase chain reaction analysis using pathologically clear (normal) or confirmed PCa tissue samples from the same patients (N=18 per group). Results: Levels (relative fold changes) of bone morphogenetic protein 7 (BMP7) transcripts were significantly lower in PCa tissues, compared with clear tissues, in a paired t-test (p=0.0075). Although neural cell adhesion molecule 1 (NCAM1) transcripts tended to be altered in PCa tissues, statistically insignificant differences were observed (p=0.0521). No statistically significant differences were observed for the other genes/miRNAs analyzed in PCa tissues due to a high degree of individual variance in expression. Conclusion: Similar to the results previously observed in rats, changes in the levels of BMP7 and NCAM1 transcripts were evident in human PCa tissues, suggesting that these genes may serve as potential diagnostic biomarkers during the early stages of PCa. Further studies are needed to determine the potential use of these molecules as biomarkers.

Objective To evaluate the long-term safety and efficacy of indacaterol/glycopyrronium (IND/GLY) in patients with COPD in a real-world setting in Japan. Methods This 52-week, multicentre, post-marketing surveillance conducted in Japan between December 2013 and August 2019 included patients using IND/GLY for the first time to relieve airway obstructive disorder-related symptoms. Safety outcomes included the incidence of adverse events (AEs), serious AEs (SAEs), adverse drug reactions (ADRs), and serious ADRs during the 52-week period. The incidence of priority variables, including cardiovascular/cerebrovascular (CCV) AEs, β-adrenergic-related or anticholinergic AEs and cough, was also assessed. Safety outcomes were also evaluated in elderly patients. Efficacy outcomes included a physician's global assessment, COPD assessment test (CAT) and lung function test. Results Of the 1,167 patients registered, 1,108 were included in the safety and efficacy analysis. In the safety analysis population, the incidence of AEs was 13.54%, that of SAEs was 4.69%, that of ADR was 3.61%, and that of serious ADRs was 0.36% over 52 weeks. CCV AEs, β-adrenergic-related and anticholinergic AEs and cough were reported as 2.62%, 1.99% and 0.63%, respectively. The physician's global assessment showed that the overall response rate at the last assessment was 74.19%. The mean (95% confidence interval) CAT scores decreased from the start of treatment to Week 52 with IND/GLY (-6.9 [-7.8 to -6.1]). The lung function (FEV1 and FVC) improved over time from the start of IND/GLY to Week 52. Conclusion IND/GLY demonstrated a good long-term safety profile in a real-world setting in Japanese patients with COPD, with beneficial effects in terms of the lung function and symptoms in clinical use.

本稿の目的は，甲南大学で筆者が担当する外国語科目（フランス語）と言語文化科目（講義科目）の授業において実践しているアクティブ・ラーニングの方法とその狙いを説明し、学習効果と今後の課題について明らかにすることである。まず、外国語科目としてのフランス語の授業のなかで筆者が実践している内容、アクティブ・ラーニングの実施が難しい文法の授業についても言及し、次に、語学演習を含まない言語文化科目において、筆者が取り入れているアクティブ・ラーニングの内容と方法について説明する。最後に、言語文化科目に関する「独自の授業実態アンケート」の結果、受講生への個別インタビューによる聞き取り調査で得られたフィードバックをもとに、こうした実践の妥当性と教育効果について検討し、改善に向けた取組みについて述べる。特に講義科目である言語文化科目において、事前学習、授業の中でのアクティブ・ラーニングへの誘い、授業後の課題への取組みを通じて得た学習体験が、近い将来、社会人として活躍するための実践的能力につながると受講生が感じていることがわかった。, Cet article a pour but d'expliquer les objectifs et les méthodes de l'« apprentissage actif » que nous pratiquons dans nos cours de langue étrangère (cours de français en l'occurrence) et dans nos cours de civilisation et de citoyenneté globale (cours magistraux) à l'Université Konan, et aussi de clarifier les effets positifs de cet apprentissage ainsi que les points à améliorer. Tout d'abord, nous présenterons les exercices pratiqués dans nos cours de français langue étrangère, et nous parlerons aussi de notre cours de grammaire, dans lequel il est difficile de mettre en oeuvre l'apprentissage actif. Ensuite, nous traiterons des méthodes d'apprentissage actif pratiquées dans le cadre de nos cours magistraux portant sur la civilisation et la citoyenneté globale, cours dans lesquels les étudiants ne font pas d'exercices linguistiques. Enfin, sur la base des résultats de questionnaires originaux effectués dans nos cours magistraux, et grâce aux réactions et aux remarques que nous avons pu recueillir à travers quelques entretiens individuels avec trois étudiants, nous nous binterrogerons sur la validité et l'efficacité de ces pratiques, et nous préciserons les points à améliorer. Nous avons pu constater qu’à travers ces cours portant sur la civilisation et la citoyenneté globale (cours magistraux) et appuyés sur les méthodes de l'apprentissage actif, les étudiants avaient le sentiment que les expériences d’apprentissage acquises par le biais de l'apprentissage préalable, des invitations à l'apprentissage actif en classe et des devoirs post-classe mèneraient à des compétences pratiques leur permettant de participer activement à la société dans un avenir proche., 大学教育におけるアクティブ・ラーニングの実践と考察 = Studies on the Implementation of Active Learning in University Classes, 欧文抄録は、p.103-104に掲載

Conjugated polymer nanoparticles (Pdots) are expected to be novel bioimaging and sensing probes. However, the size tuning required to control biological interactions has not been well established. Herein, we achieved a size-tunable synthesis of Pdots ranging from 30 to 200 nm by controlling the hydrolysis rate of the stabilising agent and evaluated their cellular imaging properties.

Intraperitoneal chemotherapy demonstrates potential applicability in the treatment of peritoneally disseminated ovarian cancer because the disseminated tumors can directly receive exposure to high concentrations of anticancer drugs. However, a considerable proportion of drugs, particularly micromolecular and hydrophilic drugs, such as cisplatin (CDDP), are often excreted through glomerular filtration for a short period. To effectively deliver CDDP into peritoneally disseminated ovarian cancer tissues, we developed an alginate (AL)-based hybrid system in which a CDDP-loaded AL nanogel (AL/CDDP-nanogel) was encapsulated in an injectable AL-hydrogel cross-linked with calcium ions. This system enabled the sustained release of CDDP from the AL/CDDP-nanogel/AL-hydrogel hybrid for over a week. Herein, we constructed a peritoneally disseminated ovarian cancer mouse model using ovarian cancer cell lines with KRAS mutations (ID8-KRAS: KRASG12V). The AL/CDDP-nanogel/AL-hydrogel hybrid system showed significant antitumor activity in vivo. This therapy may be considered a novel strategy for the treatment of advanced-stage ovarian cancer with KRAS mutations.

Recent progress in neuroscience has made it possible to use neurophysiological techniques to validate and deepen the interpretation of Rorschach variables. The aim of this article is to review the results from Rorschach studies using the neurophysiological approach to discuss the consistencies and inconsistencies between the different results, and then to consider the future direction of Rorschach research in this area. We also provide unpublished data to complement the picture from peer-reviewed studies. Two main approaches to neuropsychological studies on the Rorschach exist. One approach is to measure brain activities directly during the Rorschach administration; a series of studies using multiple neurophysiological methods revealed activation of the mirror neuron system with relation to human movement responses. Another possible approach is to investigate whether individual differences in Rorschach scores can be explained by neurophysiological measurements during the administration of another psychological task. This article reviews how these two approaches provide novel insights into the Rorschach Test.

Dans le pays de la langue de Shakespeare, le français reste, depuis longtemps et encore aujourd’hui, la première langue étrangère enseignée dans les établissements scolaires. Dans cet article, nous voudrions tout d’abord esquisser la situation actuelle de l’enseignement des langues étrangères au Royaume-Uni, et préciser ensuite certains problèmes qu’engendre le système actuel de l’enseignement du français en Angleterre par rapport aux diplômes: le « GCSE », équivalent du diplôme national du brevet français et le « GCE A-Level », équivalent du baccalauréat français. Enfin, nous aurions aimé analyser quelques manuels ainsi que des contenus des deux sortes examens en question, mais ce sera l’objet de notre prochaine étude.

The safety and efficacy of omalizumab in chronic spontaneous urticaria (CSU) patients has been established, but real-world long-term data remain scarce, especially in Japan.52-week, open-label, single-arm, observational study evaluated the safety and effectiveness of first-time omalizumab in Japanese CSU patients responding inadequately to conventional therapies.Overall, 235 of 280 patients completed the study. Most patients were aged ≥ 18 and 65 years; adolescents (≥ 12 and ≤ 18 years) accounted for 9.6% of the total population. The mean ± standard deviation (SD) duration of CSU at baseline was 1.6 ± 3.1 years; 46.1% of patients had had CSU for6 months. At baseline, the mean ± SD of Urticaria Control Test (UCT) score, Weekly Urticaria Activity Score (UAS7), and Dermatology Life Quality Index (DLQI) were 5.1 ± 3.2, 25.2 ± 11.9, and 8.4 ± 5.9, respectively. The mean ± SD duration of the observation period was 330.3 ± 86.2 days. Relapse was reported in 65 patients, 51, 9, and 5 of whom required retreatment with omalizumab 1, 2, and ≥ 3 times, respectively. The incidence of adverse events (AEs), serious AEs, and adverse drug reactions (ADRs) was reported in 11.8%, 1.4%, and 3.9% of patients, respectively. The most common AEs were urticaria (1.8%) and eczema (1.1%). No adolescents experienced ADRs. A cumulative of 92.8% of patients responded in the Physician's Global Impression of Change, with 81.3%, 75.0%, and 95.1% of patients achieving UCT ≥ 12, UAS7 ≤ 6, and DLQI ≤ 5 up to Week 52, respectively.This study supports the safety and effectiveness of omalizumab in CSU patients who responded inadequately to conventional therapies in real-world clinical practice in Japan.

Although the use of medication during pregnancy is common, information on exposure to the developing fetus and potential teratogenic effects is often lacking. This study used a rat model to examine the placental transfer of three small-molecule drugs with molecular weights ranging from approximately 300 to 800 Da with different physicochemical properties. Time-mated Sprague Dawley (Hsd:SD) rats aged 11-13 weeks were administered either glyburide, rifaximin, or fentanyl at gestational day 15. Maternal blood, placentae, and fetuses were collected at 5 min, 30 min, 1 h, 4 h, 8 h, 24 h, 48 h, and 96 h post-dose. To characterize the rate and extent of placental drug transfer, we calculated several pharmacokinetic parameters such as maximum concentration (Cmax), time to maximum concentration (Tmax), area under the concentration-time curve (AUC), half-life (t

Abstract. This article presents a Therapeutic Assessment (TA; Finn, 2007 ) case study of a teenage girl assessed because of a complex set of symptoms: history of suicidal statements, sensory sensitivity, questions about gender identity, depression, anxiety/OCD, and possible subclinical psychotic symptoms. Logan’s parents, Joe and Susan, participated in the assessment, with Susan attending every appointment and Joe attending only the first and last. Ten instruments were administered, with each providing information about different aspects of Logan. The Thurston Cradock Test of Shame (TCTS; Thurston & Cradock O’Leary, 2009 ) scores and analysis encompassed all of the other test findings, clarified details not fully explained by tests such as the Rorschach and MMPI-A, and brought the core adolescent and family issue of shame to the foreground. When the assessors explained shame to Logan’s mother, she was better able to understand her daughter’s shame and connect with her own feelings of shame. When Logan’s father arrived for the final feedback session, he was quite tense, argumentative, and threatened to cancel the appointment. The TCTS allowed the assessors to understand Joe’s aggressive behavior as signs of TCTS defenses of deflation, aggression, and inflation/contempt, appreciate his underlying feelings of shame, and to delicately restore the “interpersonal bridge” ( Kaufman, 1996 ), which is key to reducing shame. As a result, Joe’s defensiveness decreased, and he was open to continuing the feedback session. In the end, Joe was able to identify with his daughter in a new way and even admit his own sense of shame in the session. This provided an opportunity for healing in the whole family.

The authors examined the effect of interdependent group-oriented contingencies on reducing the preparation time for the snack time. The study was conducted in a club for after-school activities for children with 37 first- to sixth-grade students.The research period was from March 6, 2018 to March 31, 2018. The authors observed and recorded 14 sessions: Sessions 1-5 were the baseline phase, 6-10, the intervention phase, and the final 11-14, the follow-up phase. After the 5th session, a target behavior was set for all the children for sessions 6-10, and a staff member announced that he would record how long the children took to prepare for their snack time. We also recorded the number of times the serving group advised them to hurry up.During the intervention period and the follow-up period the time for preparing for their snack time was shortened and the number of times the serving group advised them to hurry up decreased.

The imbalance of testosterone to estradiol ratio has been related to the development of prostate diseases. Although rat models of prostate diseases induced by endocrine-disrupting chemicals (EDCs) and/or hormone exposure are commonly used to analyze gene expression profiles in the prostate, most studies utilize a single endpoint. In this study, microarray analysis was used for gene expression profiling in rat prostate tissue after exposure to EDCs and sex hormones over multiple time points (prepubertal through adulthood). We used dorsolateral prostate tissues from Sprague-Dawley rats (male offspring) and postnatally administered estradiol benzoate (EB) on postnatal days (PNDs) 1, 3, and 5, followed by treatment with additional hormones [estradiol (E) and testosterone (T)] on PNDs 90-200, as described by Ho et al. Microarray analysis was performed for gene expression profiling in the dorsolateral prostate, and the results were validated via qRT-PCR. The genes in cytokine-cytokine receptor interaction, cell adhesion molecules, and chemokines were upregulated in the EB+T+E group on PNDs 145 and 200. Moreover, early-stage downregulation of anti-inflammatory gene: bone morphogenetic protein 7 gene was observed. These findings suggest that exposure to EB, T, and E activates multiple pathways and simultaneously downregulates anti-inflammatory genes. Interestingly, these genes are reportedly expressed in prostate cancer tissues/cell lines. Further studies are required to elucidate the mechanism, including analyses using human prostate tissues.

A 47-year-old male with macroglossia presented with dyspnea on effort and chest pain at rest. Cardiac MRI revealed diffuse global subendocardial late gadolinium enhancement below the left ventricular endocardium and a dark blood pool of intracardiac contrast medium. Tongue biopsy revealed amyloid deposition, which was limited in the myocardium. He was diagnosed with primary light chain amyloidosis. His condition was stage I according to the Mayo Clinic staging system. He underwent autologous peripheral blood stem cell transplantation. On Day 10, he developed chest pain and died suddenly on Day 11. Postmortem examination revealed amyloid deposition throughout the heart.

Objective: To evaluate the long-term safety and efficacy of glycopyrronium (GLY) in patients with COPD in a real-world setting in Japan. Methods: This 52-week, multicentre, post-marketing surveillance conducted in Japan, between February 2013 and August 2019, included patients using GLY for the first time for the relief of airway obstructive disorder-related symptoms. Safety outcomes included incidence of adverse events (AEs), serious AEs (SAEs), adverse drug reactions (ADRs), serious ADRs (SADRs) and priority variables included cardiovascular/cerebrovascular (CCV) AEs and anticholinergic AEs during the 52-week period. Safety outcomes were also assessed in elderly patients. Efficacy outcomes included physician’s global assessment, COPD assessment test (CAT) and lung function test. Results and Discussion: Of the 1,331 patients registered for this surveillance, safety and efficacy outcomes were evaluated in 1,277 patients. In the safety analysis population, the incidence of AEs was 15.51%, SAEs 4.70%, ADRs 5.01% and SADRs 0.31%. The CCV AEs and anticholinergic AEs were reported by 0.70% and 2.58% patients, respectively. Physician’s global assessment showed that the overall response rate at the last assessment was 70%. The mean (95% CI) CAT scores decreased from the start of treatment to Week 52 with GLY, (−6.2 [−7.0 to −5.4]). Lung function in terms of trough FEV1 and FVC improved over time from the start of GLY to Week 52. Conclusion: GLY demonstrated an acceptable long-term safety profile with no new safety concerns in a real-life setting. It demonstrated improvement in lung function and symptom control in Japanese COPD patients.

The focus of this brief review is to describe the application of nanoparticles, including endogenous nanoparticles (e.g., extracellular vesicles, EVs, and virus capsids) and exogenous nanoparticles (e.g., organic and inorganic materials) in cancer therapy and diagnostics. In this review, we mainly focused on EVs, where a recent study demonstrated that EVs secreted from cancer cells are associated with malignant alterations in cancer. EVs are expected to be used for cancer diagnostics by analyzing their informative cargo. Exogenous nanoparticles are also used in cancer diagnostics as imaging probes because they can be easily functionalized. Nanoparticles are promising targets for drug delivery system (DDS) development and have recently been actively studied. In this review, we introduce nanoparticles as a powerful tool in the field of cancer therapy and diagnostics and discuss issues and future prospects.

Rats are the standard model for male reproductive toxicity testing. Rat prostates are physiologically and anatomically different from those of humans. Drug and chemical toxicity testing would benefit from an in vitro model of human prostate cells. Recently, spheroids derived by three-dimensional culture of human cell lines have been used for assessing drug and chemical toxicity in vitro as they mimic in vivo environments more closely than two-dimensional culture. However, forming consistently sized, uniform spheroids is technically challenging for toxicity testing. The purpose of this study was to identify potential genetic markers for assessing prostatic toxicity in spheroids. We formed prostate spheroids using agarose-coated plates seeded with human primary prostate epithelial cells. Prostate spheroids were treated with either 17β-estradiol (E2) or testosterone (T) on days 2-7 of culture. Samples were harvested on culture day 7. qPCR was used to examine gene expression levels previously identified in rats with chronic inflammation exposed to estradiol benzoate, E2 and/or T. Changes in some gene expression levels were observed in the spheroids treated with E2 or T. We found that treatment with 1 nM E2 and/or 10 μM T significantly altered spheroid proliferation and viability, as well as the expression levels of genes including Nanog homeobox (NANOG), C-C motif chemokine ligand 2 (CCL2) and bone morphogenetic protein receptor type 2 (BMPR2). Further studies using biologically active molecules with prostatic toxicity are needed to verify the results and to determine whether gene expression changes in the spheroid are specific to E2 or T treatment.

Intraperitoneal chemotherapy demonstrates potential applicability in the treatment of peritoneally disseminated ovarian cancer because the disseminated tumors can directly receive exposure to high concentrations of anticancer drugs. However, a considerable proportion of drugs, particularly micromolecular and hydrophilic drugs, such as cisplatin (CDDP), are often excreted through glomerular filtration for a short period. To effectively deliver CDDP into peritoneally disseminated ovarian cancer tissues, we developed an alginate (AL)-based hybrid system in which a CDDP-loaded AL nanogel (AL/CDDP-nanogel) was encapsulated in an injectable AL-hydrogel cross-linked with calcium ions. This system enabled the sustained release of CDDP from the AL/CDDP-nanogel/AL-hydrogel hybrid for over a week. Herein, we constructed a peritoneally disseminated ovarian cancer mouse model using ovarian cancer cell lines with KRAS mutations (ID8-KRAS: KRAS

Carboxymethyl cellulose (CMC) is a promising material for moist wound healing, and silver loading onto CMC has been examined for anti-bacterial activity. In this study, we developed silver-loaded CMC nonwoven sheets with different counterions, namely sodium CMC (CMC-Na/Ag) and partially protonated CMC (CMC-H/Ag), to examine their anti-bacterial and wound-healing properties. Owing to the presence of counter protons, CMC-H/Ag showed slower water adsorption, dissolution, and Ag release than CMC-Na/Ag. In addition, CMC-H/Ag and CMC-Na/Ag exhibited differences in anti-bacterial activities in shake-flask and inhibition zone tests in vitro. An in vivo experiment using a pressure ulcer mouse model with Pseudomonas aeruginosa infection showed that CMC-Na/Ag significantly accelerated wound healing compared to CMC-H/Ag and a commercially available Ag-loaded CMC nonwoven sheet, Aquacel Ag. These results suggest the importance of controlling CMC counterions and the therapeutic potential of the developed product as a wound dressing.

ObjectivesTo analyse the clinical characteristics and prognosis of acute exacerbation (AE) in patients with idiopathic pulmonary fibrosis (IPF) and pulmonary emphysema.DesignA multicentre retrospective cohort studySettingTwo university hospitals in JapanParticipantsPatients admitted to hospitals due to AE of IPF diagnosed based on a multidisciplinary discussion.InterventionsNonePrimary and secondary outcome measures90-day mortality rateMethodsWe retrospectively analysed consecutive patients with AE of IPF, with or without pulmonary emphysema, admitted to two university hospitals between 2007 and 2018.ResultsAmong 62 patients (median age, 75 years; 48 men) admitted for AE of IPF, 29 patients (46%) presented with concomitant pulmonary emphysema. There was no significant difference in the arterial partial oxygen pressure/fraction of inhaled oxygen (P/F) ratio or other laboratory and radiographic data between patients with and without emphysema. The 90-day mortality rate was significantly lower in patients with emphysema than in those with IPF alone (23% vs 52%, p=0.03). The median survival time was significantly longer in patients with emphysema than in those with IPF alone (405 vs 242 days, p=0.02).ConclusionPatients with IPF and emphysema had better short-term survival after AE than those with non-emphysematous IPF.

Background There is a need to develop in vitro models to test drugs and chemicals that induce toxicity in the male reproductive system. We have evaluated an in vitro mouse testis organ culture model capable of producing viable, fertilization-proven sperm as a possible toxicity test model. Although this in vitro model was limited to round spermatid differentiation, histopathology observations could still be performed. Liquid chromatography/mass spectrometry analysis (LC/MS)-based metabolomics was used to measure metabolome changes of chemically treated in vitro testis fragments. Methods On Postnatal Day 5, C57BL/6J mouse testes were divided into four fragments, which were placed onto a 1.5% agarose gel cube and cultured in α-MEM including 0.4% AlbuMAX I (Day 0). On Day 1 of culture, testis fragments were treated with 0 (control), 0.01, or 1 nM ethinylestradiol (EE). On Day 20 of culture, the testis fragments were collected for LC/MS and histology analysis. Results Several metabolites involved in glycogen metabolism and glycolysis pathways (uridine diphosphate-glucose, glucose phosphate, and pyruvate), in the tricarboxylic acid cycle pathway (oxaloacetate and aspartate), and in the arginine and proline metabolism (arginine and spermine) were significantly altered in the 1 nM EE treated group compared to the control group. The metabolite changes were associated with an increase in percentage of seminiferous tubules with round spermatids as well as dose-dependent dead cells. Conclusion These findings suggest that EE treatment may cause testicular toxicity by affecting glycogen metabolism and energy pathways. To confirm these findings, further experiments will be necessary using other testicular toxicants.

Background Previously, we evaluated optimal organ culture conditions to produce elongated spermatids in an in vitro mouse testis culture system. However, differences in testicular function between the cultured testis fragments and animal testis have not been determined. Methods To examine how closely cultured testis fragments in vitro approximates what typically occurs during the first wave of spermatogenesis in vivo, C57BL/6J mouse testis fragments obtained on postnatal day (PND) 5 were cultured in AlbuMAX™ I/ α-Minimal Essential Medium for 15, 23, 30, 35, 42, and 49 days, and compared to mouse testes obtained at PND 5, 14, 20, 24, 28, 30, 35, and 40. At the specified days of culture or PND of mice, the following analyses were conducted: histology, flow cytometry for haploid cell detection, qPCR for spermatid markers, and liquid chromatography/mass spectrometry for testosterone levels. Results Round spermatids were initially observed at 23 days, and their percentage of the total number of cells continued to increase with culture time, as did gene expression of the spermatid markers and haploid cell percentage in the cultured testis fragments. These results were similar in temporal sequence to those in animals. Testosterone levels in the testis fragments reached a maximum at Day 49. Conclusion These findings show this in vitro mouse testis organ culture model may be a useful and convenient tool for mechanistic studies. However, because germ cell differentiation in all seminiferous tubules was not observed, improvements in the system/methods are needed to more closely replicate spermatogenesis as observed in animals.

The assessment of xenobiotic-induced testicular toxicity is important in drug development. Nonetheless, in vitro models to test drugs and chemicals that may cause testicular toxicity are lacking, requiring the continued use of animal models for those studies. We previously evaluated an in vitro mouse testis organ culture system using ethinylestradiol (EE), a well-studied testicular toxicant, and demonstrated a dose-dependent relationship between adverse effects to germ cell differentiation and increasing EE concentrations. However, we terminated that study after 20 days of culture due to oxygen deficiency during germ cell differentiation. Therefore, in the current study, we aimed to identify gene(s) with potential for supporting the histopathological evaluations of testicular toxicity using in vitro testis organ culture system. We cultured testis fragments obtained from mice at postnatal day (PND) 5 in α-Minimal Essential Medium containing 40 mg/mL AlbuMAX™ I and treated them with 0.01 or 1 nM EE on day 1 of culture. On day 20, we collected testis fragments for RNA sequencing analysis and quantitative polymerase chain reaction (qPCR). We found that phospholipase C, zeta 1 and testis-specific serine kinase 4 genes, that are involved in spermatogenesis and predominantly expressed in the testis, were significantly reduced in testis fragments treated with the highest concentration of EE. Also, cytochrome P450, family 26, subfamily b, polypeptide 1 (Cyp26b1) and interleukin 16 (Il16) were up-regulated in the highest EE-treated groups. Further studies are needed to confirm the variations of these gene expression using other testicular toxicants.