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Germ free

Sellappan Selvaraju, Sivashanmugam Parthipan, Lakshminarayana Somashekar, B Krishnan Binsila, Atul P Kolte, Arunachalam Arangasamy, Janivara Parameshwaraiah Ravindra, Stephen A Krawetz
With artificial insemination (AI) and other precision dependent assisted reproductive technologies (ART) being followed in large scale in human and animal reproduction, assessing semen quality and fertilizability is under continuous scrutiny. Various tests have been developed to predict semen quality, but so far no single, highly reliable test is available. In this regard, transcriptomic profiling of spermatozoa assumes significance as it carries the information about spermatogenesis, sperm function, and paternal roles in post-fertilization events...
March 14, 2018: Systems Biology in Reproductive Medicine
Naoki Sawada, Takenori Kotani, Tasuku Konno, Jajar Setiawan, Yuka Nishigaito, Yasuyuki Saito, Yoji Murata, Ken-Ichi Nibu, Takashi Matozaki
In the mouse olfactory bulb (OB), interneurons such as granule cells and periglomerular cells are continuously replaced by adult-born neurons, which are generated in the subventricular zone (SVZ) of the brain. We have now investigated the role of commensal bacteria in regulation of such neuronal cell turnover in the adult mouse brain. Administration of mixture of antibiotics to specific pathogen-free (SPF) mice markedly attenuated the incorporation of bromodeoxyuridine (BrdU) into the SVZ cells. The treatment with antibiotics also reduced newly generated BrdU-positive neurons in the mouse OB...
March 9, 2018: Biochemical and Biophysical Research Communications
Youngji Cho, Galeb Abu-Ali, Hiroki Tashiro, David I Kasahara, Traci A Brown, Jeffrey D Brand, Joel A Mathews, Curtis Huttenhower, Stephanie A Shore
Previous reports demonstrate that the microbiome impacts allergic airway responses, including airway hyperresponsiveness, a characteristic feature of asthma. Here we examined the role of the microbiome in pulmonary responses to a non-allergic asthma trigger, ozone. We depleted the microbiota of conventional mice with either a single antibiotic (ampicillin, metronidazole, neomycin, or vancomycin) or a cocktail of all four antibiotics given via the drinking water. Mice were then exposed to room air or ozone. In air-exposed mice, airway responsiveness was not different in antibiotic- versus control water-treated mice...
March 12, 2018: American Journal of Respiratory Cell and Molecular Biology
Kan Gao, Yu Pi, Chun-Long Mu, Yu Peng, Zan Huang, Wei-Yun Zhu
The evidence of gut microbiota-mediated modulation of brain function has been widely recognized from studies using germ-free rodents or animals with oral antibiotic-induced microbiota depletion. Since the number of bacteria in the large intestine greatly exceeds that found within the small intestine, large intestinal microbiota may play a crucial role in the modulation of brain function. In the present study, twelve piglets (12.08 ± 0.28 kg) fitted with a T-cannula at the distal ileum were fed a standard diet and randomly assigned to two groups (n=6) for ileal infusion of either saline (control group) or antibiotics (antibiotic group)...
March 10, 2018: Journal of Neurochemistry
E A Sogorin, S Ch Agalarov
The translation of uncapped mRNAs encoding luciferase and green fluorescent protein in a cell-free translation system based on wheat germ extract has been studied. It turned out that two simultaneously translated (in one tube) different templates in a certain range of concentrations not only do not compete, but mutually enhance each other's translation. It has been shown that the synthesis of luciferase in the presence of mRNA that encodes green fluorescent protein is much more effective than in the translation of only luciferase mRNA at the same concentration...
January 2018: Molekuliarnaia Biologiia
Briony K Varda, Patricia Cho, Andrew A Wagner, Richard S Lee
BACKGROUND: In adult urologic oncology the use of robotics has become commonplace; in pediatric urology it is rare. Herein, we describe a collaboration between an adult and a pediatric urologist performing robotic surgery for children and young adults with suspicious or cancerous genitourinary (GU) lesions. OBJECTIVES: To evaluate clinical and oncologic outcomes in children and young adults undergoing robotic surgery for suspicious or cancerous lesions of the GU tract; to describe our collaborative model between an adult and pediatric surgeon at a free-standing children's hospital...
March 2, 2018: Journal of Pediatric Urology
Alla Splichalova, Vera Slavikova, Zdislava Splichalova, Igor Splichal
Preterm infants born with immature organ systems, which can impede normal development, can also be highly sensitive to different biological and/or environmental factors. Animal models could aid in investigating and understanding the effects of different conditions on the health of these immunocompromised infants. The epitheliochorial placentation of the pig prevents the prenatal transfer of protective colostral immunoglobulins. Surgical colostrum-deprived piglets are free of maternal immunoglobulins, and the cells that are normally provided via colostrum...
2018: Frontiers in Immunology
Alexander Rodriguez-Palacios, Natalia Aladyshkina, Jessica C Ezeji, Hailey L Erkkila, Mathew Conger, John Ward, Joshua Webster, Fabio Cominelli
Germ-Free (GF) research has required highly technical pressurized HEPA-ventilation anchored systems for decades. Herein, we validated a GF system that can be easily implemented and portable using Nested Isolation (NesTiso). GF-standards can be achieved housing mice in non-HEPA-static cages, which only need to be nested 'one-cage-inside-another' resembling 'Russian dolls'. After 2 years of monitoring ~100,000 GF-mouse-days, NesTiso showed mice can be maintained GF for life (>1.3 years), with low animal daily-contamination-probability risk (1 every 867 days), allowing the expansion of GF research with unprecedented freedom and mobility...
February 28, 2018: Scientific Reports
Lisbeth Charlotte Olsen, Ioannis Kourtesis, Henriette Busengdal, Marit Flo Jensen, Harald Hausen, Daniel Chourrout
BACKGROUND: Germ cell formation has been investigated in sessile forms of tunicates. This process involves the release of a subset of maternal transcripts from the centrosome-attracting body (CAB) in the progenitor cells of the germ line. When germ-soma segregation is completed, CAB structures are missing from the newly formed primordial germ cells (PGCs). In free-swimming tunicates, knowledge about germ cell formation is lacking. In this investigation, comparative gene expression and electron microscopy studies were used to address germ cell formation in Oikopleura dioica (O...
February 27, 2018: BMC Developmental Biology
Marta Trevisan, Vanessa Barbaro, Silvia Riccetti, Giulia Masi, Luisa Barzon, Patrizia Nespeca, Gualtiero Alvisi, Enzo Di Iorio, Giorgio Palù
Transgene free UNIPDi002-A-human induced pluripotent stem cell (hiPSC) line was generated by Sendai Virus Vectors reprogramming from human oral mucosal epithelial stem cells (hOMESCs) of a patient affected by ectrodactyly-ectodermal dysplasia-clefting (EEC)-syndrome, carrying a mutation in exon 8 of the TP63 gene (R304Q). The UNIPDi002-A-hiPSC line retained the mutation of the parental R304Q-hOMESCs and displayed a normal karyotype. No residual expression of transgenes nor Sendai virus vector sequences were detected in the line at passage 8...
February 16, 2018: Stem Cell Research
Michal Chovanec, Zuzana Cierna, Viera Miskovska, Katarina Machalekova, Katarina Kalavska, Katarina Rejlekova, Daniela Svetlovska, Dusan Macak, Stanislav Spanik, Karol Kajo, Pavel Babal, Ugo De Giorgi, Michal Mego, Jozef Mardiak
BACKGROUND: We evaluated systemic immune-inflammation index (SII) and its association with patient outcome in germ-cell tumours (GCTs). METHODS: Two independent cohorts of patients were analysed; the discovery set (n=171) from a single institution and the validation set (n=181) previously included in a study evaluating PD-L1 in GCTs. The SII was calculated using platelet (P), neutrophil (N) and lymphocyte (L) counts before chemotherapy and correlated with survival using regression analyses and Kaplan-Meier method...
February 27, 2018: British Journal of Cancer
Jikang Wu, Anice Sabag-Daigle, Mikayla A Borton, Linnea F M Kop, Blake E Szkoda, Brooke L Deatherage Kaiser, Stephen R Lindemann, Ryan S Renslow, Siwei Wei, Carrie D Nicora, Karl K Weitz, Young-Mo Kim, Joshua N Adkins, Thomas O Metz, Prosper Boyaka, Venkat Gopalan, Kelly C Wrighton, Vicki H Wysocki, Brian M M Ahmer
Salmonella enterica elicits intestinal inflammation to gain access to nutrients. One of these nutrients is fructose-asparagine (F-Asn). The availability of F-Asn to Salmonella during infection is dependent upon the pathogenicity islands, SPI1 and SPI2, which in turn are required to provoke inflammation. Here, we determined that F-Asn is present in mouse chow at approximately 400 pmol/mg dry weight. F-Asn is also present in the intestinal tract of germ-free mice at 2700 pmol/mg dry weight, and in the intestinal tract of conventional mice at 15 pmol/mg...
February 26, 2018: Infection and Immunity
Xianfa Yang, Ran Wang, Xiongjun Wang, Guoqing Cai, Yun Qian, Su Feng, Fangzhi Tan, Kun Chen, Ke Tang, Xingxu Huang, Naihe Jing, Yunbo Qiao
Clinical therapies of pluripotent stem cells (PSCs)-based transplantation have been hindered by frequent development of teratomas or tumors in animal models and clinical patients. Therefore, clarifying the mechanism of carcinogenesis in stem cell therapy is of great importance for reducing the risk of tumorigenicity. Here we differentiate Oct4-GFP mouse embryonic stem cells (mESCs) into neural progenitor cells (NPCs) and find that a minority of Oct4+ cells are continuously sustained at Oct4+ state. These cells can be enriched and proliferated in a standard ESC medium...
February 22, 2018: Journal of Molecular Cell Biology
Suyatno, Yuka Kitamura, Shuntaro Ikeda, Naojiro Minami, Masayasu Yamada, Hiroshi Imai
Undifferentiated spermatogonia eventually differentiate in the testis to produce haploid sperm. Within this cell population, there is a small number of spermatogonial stem cells (SSCs). SSCs are rare cells in the testis, and their cellular characteristics are poorly understood. Establishment of undifferentiated cell line would provide an indispensable tool for studying their biological nature and spermiogenesis/spermatogenesis in vitro. However, there have been few reports on the long-term culture of undifferentiated spermatogonia in species other than rodents...
January 17, 2018: Molecular Reproduction and Development
Charline Jansch, Katharina Günther, Jonas Waider, Georg C Ziegler, Andrea Forero, Sina Kollert, Evgeniy Svirin, Dirk Pühringer, Chee Keong Kwok, Reinhard Ullmann, Anna Maierhofer, Julia Flunkert, Thomas Haaf, Frank Edenhofer, Klaus-Peter Lesch
Fibroblasts were isolated from a skin biopsy of a clinically diagnosed 51-year-old female attention-deficit/hyperactivity disorder (ADHD) patient carrying a duplication of SLC2A3, a gene encoding neuronal glucose transporter-3 (GLUT3). Patient fibroblasts were infected with Sendai virus, a single-stranded RNA virus, to generate transgene-free human induced pluripotent stem cells (iPSCs). SLC2A3-D2-iPSCs showed expression of pluripotency-associated markers, were able to differentiate into cells of the three germ layers in vitro and had a normal female karyotype...
February 13, 2018: Stem Cell Research
Brendan A Daisley, Mark Trinder, Tim W McDowell, Stephanie L Collins, Mark W Sumarah, Gregor Reid
Despite benefits to the global food supply and agricultural economies, pesticides are believed to pose a threat to both human and wildlife health. Chlorpyrifos (CP), a commonly used organophosphate insecticide, has poor target-specificity and causes acute neurotoxicity in a wide range of species via suppression of acetylcholinesterase. This effect is exacerbated 10- to 100-fold by chlorpyrifos-oxon (CPO), a principal metabolite of CP. Since many animal-associated symbiont microorganisms are known to hydrolyze CP into CPO, we used a Drosophila melanogaster insect model to investigate the hypothesis that indigenous and probiotic bacteria could affect CP metabolism and toxicity...
February 23, 2018: Applied and Environmental Microbiology
F Yilmaz, N Soyer, R Uslu, A P Erdogan, B Karaca, G Saydam, F Sahin, F Vural
BACKGROUND AND AIM: About 20-25% of the testicular germ cell tumors (TGCT) are relapsed or refractory after first line therapy and optimal treatment for this group is poorly defined. We aimed to analyze the efficacy and safety of autologous stem cell transplantation (ASCT) in this patient group.Material and. METHODS: 19 patients with 28 ASCT were retrospectively analyzed. All the patients were treated with BEP (Bleomycin, etoposide, cisplatin) as first line therapy and TIP(paclitexalifosfamide, cisplatin) was given as salvage chemotherapy...
April 2017: Indian Journal of Cancer
Yasuyuki Suzuki, Tomio Ogasawara, Yuki Tanaka, Hiroyuki Takeda, Tatsuya Sawasaki, Masaki Mogi, Shuang Liu, Kazutaka Maeyama
G-protein-coupled receptors (GPCRs) are membrane proteins distributed on the cell surface, and they may be potential drug targets. However, synthesizing GPCRs in vitro can be challenging. Recently, some cell-free protein synthesis systems have been shown to produce a large amount of membrane protein combined with chemical chaperones that include liposomes and glycerol. Liposomes containing high concentrations of glycerol are known as glycerosomes, which are used in new drug delivery systems. Glycerosomes have greater morphological stability than liposomes...
2018: Frontiers in Pharmacology
Mona Mischke, Tulika Arora, Sebastian Tims, Eefje Engels, Nina Sommer, Kees van Limpt, Annemarie Baars, Raish Oozeer, Annemarie Oosting, Fredrik Bäckhed, Jan Knol
AIMS: The metabolic state of human adults is associated with their gut microbiome. The symbiosis between host and microbiome is initiated at birth, and early life microbiome perturbation can disturb health long-lastingly. Here, we determined how beneficial microbiome interventions in early life affect metabolic health in adulthood. METHODS: Postnatal diets were supplemented with either prebiotics (scGOS/lcFOS) or synbiotics (scGOS/lcFOS with Bifidobacterium breve M-16V) until post-natal (PN) day 42 in a well-established rodent model for nutritional programming...
February 20, 2018: Diabetes, Obesity & Metabolism
Erica E Alexeev, Jordi M Lanis, Daniel J Kao, Eric L Campbell, Caleb J Kelly, Kayla D Battista, Mark E Gerich, Brittany R Jenkins, Seth T Walk, Douglas J Kominsky, Sean P Colgan
Interactions between the gut microbiota and the host are important for health, where dysbiosis has emerged as a likely component of mucosal disease. The specific constituents of the microbiota that contribute to mucosal disease are not well defined. We sought to define microbial components that regulate homeostasis within the intestinal mucosa. Using an unbiased, metabolomic profiling approach, a selective depletion of indole and indole-derived metabolites was identified in murine and human colitis. Indole-3-propionic acid (IPA) was selectively diminished in circulating serum from human subjects with active colitis and that IPA served as a biomarker of disease remission...
February 15, 2018: American Journal of Pathology
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