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Background and objectives Transdermal differentiation of human adipose stem cells (ASCs) to other cell types is still a challenge in regenerative medicine. Studies using primary ASCs are also limited as they may undergo replicative senescence during repeated passages in vitro. However, ASCs immortalized (iASCs) with human telomerase enzyme expressing plasmid exhibits a uniform population suitable for differentiation in vitro. A right combination of biological and physical stimuli may induce transdermal differentiation of iASCs into neurons in vitro. Study design/materials and methods iASCs were differentiated to free-floating neural stem cell aggregates (neurospheres) using a combination of growth inducers. Cells in these spheres were induced to differentiate into neurons using low-intensity lasers by a process called photobiomodulation (PBM). Results Laser at the near infrared (NIR) wavelength 825 nm and fluences 5, 10, and 15 J/cm2 was capable of increasing the differentiation of neurospheres to neurons. Precisely, there was a statistically significant increase in the early neuronal marker at 5 J/cm2 and a much appreciable increase at 15 J/cm2 in correlation with the biphasic dose response of PBM. However, these differentiated cells failed to express late neuronal markers in vitro. Comparison of these differentiating iASCs with the primary ASCs revealed a sharp distinction between the metabolic processes of the primary ASCs, neurospheres, and newly differentiated neurons. Conclusion We found that PBM increased the yield of neurons and effected stem cell differentiation through modulation of cellular metabolism and redox status. Our study also identifies that iASCs are an excellent model for analysis of stem cell biology and for performing transdermal differentiation. Significance This study demonstrates that a combination of biological and physical inducers can advance the differentiation of adipose stem cells to neurons. We were able to establish the optimal energy for the neuronal differentiation of iASCs in vitro. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Adipose Stem Cells (ASCs) are capable of neuronal differentiation, which makes them an ideal choice for therapies in nerve injuries. Principally, the differentiation of autologous ASCs to neurons offers solutions for the replacement therapies of nervous system with patient's own genetic background. On the contrary, the use of genetically modified (immortalized) ASCs has the benefit of accessibility by surpassing ethical concerns and ease for propagation as a continuous cell culture. Photobiomodulation (PBM) is a therapeutic modality with laser or light, which is widely been used for modulating stem cell bioprocesses viz. proliferation and differentiation. A comparative analysis was performed to evaluate the neuronal differentiation potential of primary ASCs isolated from a healthy human subject with commercially obtained immortalized ASCs with PBM. The outcome of this analysis will help us to know either primary or immortalized ASCs are most suitable for biomedical applications. Both primary and immortalized ASCs were characterized using their surface protein markers CD44/90/133/166 and induced to differentiate into neuronal cells using Fibroblast Growth Factor, basic (bFGF) and forskolin following PBM using Near Infra-Red (NIR) lasers. Based on the expression of nestin, an early neuronal marker an exposure to 5, 10 and 15 J/cm

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Additional file of Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

The promise of engineering specific cell types from stem cells and rebuilding damaged or diseased tissues has fascinated stem cell researchers and clinicians over last few decades. Mesenchymal Stem Cells (MSCs) have the potential to differentiate into non-mesodermal cells, particularly neural-lineage, consisting of neurons and glia. These multipotent adult stem cells can be used for implementing clinical trials in neural repair. Ongoing research identifies several molecular mechanisms involved in the speciation of neuroglia, which are tightly regulated and interconnected by various components of cell signalling machinery. Growing MSCs with multiple inducers in culture media will initiate changes on intricately interlinked cell signalling pathways and processes. Net result of these signal flow on cellular architecture is also dependent on the type of ligands and stem cells investigated in vitro. However, our understanding about this dynamic signalling machinery is limited and confounding, especially with spheroid structures, neurospheres and organoids. Therefore, the results for differentiating neurons and glia in vitro have been inconclusive, so far. Added to this complication, we have no convincing evidence about the electrical conductivity and functionality status generated in differentiating neurons and glia. This review has taken a step forward to tailor the information on differentiating neuroglia with the common methodologies, in practice.

Irradiation with red light or near-infrared (NIR) lasers can bio-modulate cellular processes or revitalize injured tissues and therefore, widely been used for therapeutic interventions. Mechanistically, this cellular or biological process, referred as Photobiomodulation (PBM), is achieved by the generation of oxide free radicals in cells and tissues. This explorative study using red light (636 nm) and Near Infra-Red (NIR, 825 nm) laser at various irradiation exposures reckons the level of oxidative stress induced by these free radicals in human primary fibroblasts. Freshly isolated dermal fibroblasts were irradiated with red light and NIR at power densities of 74 and 104 mV/cm2, respectively and, at varying fluences ranging from 5 to 25 J/cm2. Cellular oxidative stress, measured by Reactive Oxygen Species (ROS) upon quantifying fluorescently labelled oxide free radicals in cells, detected considerable variations between the irradiation exposures of red light and NIR laser. The NIR laser demonstrated high levels of ROS at all fluences, except 10 J/cm2 indicating its ability in generating of two types of oxide radicals in dermal fibroblasts, often illustrated as biphasic response. Further, the responses of these cells to variable fluences of red light and NIR laser were measured to evaluate the immediate effect of PBM on cellular activity. The production of cellular energy coincides with the amount of oxidative stress, which was two-fold higher in cells irradiated with the NIR laser, as compared with the red light. This outcome indicates that the ROS production within biological systems are more dependent on the wavelength of the laser rather than its fluences. Further studies are required to avoid ‘overdosing of PBM’ and to analyse ROS qualitatively for making the best use of the red light and NIR laser in clinics.

Neurological diseases and disorders pose a challenge for treatment and rehabilitation due to the limited capacity of the nervous system to repair itself. Adipose stem cells (ASCs) are more pliable than any adult stem cells and are capable of differentiating into non-mesodermal tissues, including neurons. Transdifferentiating ASCs to specific neuronal lineage cells enables us to deliver the right type of cells required for a replacement therapy into the nervous system.Several methodologies are being explored and tested to differentiate ASCs to functional neurons and glia with cellular factors and chemical compounds. However, none of these processes and prototypes has been wholly successful in changing the cellular structure and functional status of ASCs to become identical to neuroglial cells. In addition, successful integration and functional competence of these cells for use in clinical applications remain problematic. Photobiomodulation or low-level laser irradiation has been successfully applied to not only improve ASC viability and proliferation but has also shown promise as a possible enhancer of ASC differentiation.Studies have shown that photobiomodulation improves the use of stem cell transplantation for neurological applications. This review investigates current neuro-differentiation inducers and suitable methodologies, including photobiomodulation, utilizing ASCs for induction of differentiation into neuronal lineages.

An efficient and scalable synthesis of AZD3264 is described in which the differential reactivities of various halogen atoms have been employed. The process involves five linear chemical steps with three isolated stages starting from commercially available fragments.

Antibiotics such as chlortetracycline (CTC) have been used to promote growth of pigs for decades, but concerns over increased antibiotic-resistant infections in humans have prompted the development of alternative strategies. Developing alternatives to antibiotic growth promoters (AGPs) could be informed by information on the mechanisms of growth promotion, notably, how AGPs affect the microbial populations of the gastrointestinal tract. Pigs from three sows were aseptically delivered by cesarean section. Six piglets were distributed to each of two foster mothers until weaning, when piglets were fed a diet with or without 50 mg/kg CTC for 2 weeks. The ileal bacterial microbiota was characterized by using a cultivation-independent approach based on DNA extraction, PCR amplification, cloning, and sequencing of the 16S rRNA gene pool. The ileal and mucosal communities of these growing pigs were dominated by Lactobacillus bacteria, various members of the family Clostridiaceae , and members of the poorly known genus Turicibacter . Overall, CTC treatment resulted in three shifts: a decrease in L actobacillus johnsonii , an increase in L . amylovorus , and a decrease in Turicibacter phylotypes. The composition of the microbiota varied considerably between individual pigs, as revealed by shared operational taxonomic units (OTUs) and similarity (SONS) analysis (θ YC values). While the observed variation between untreated pigs obscured the possible effect of CTC, ∫-LIBSHUFF and SONS analyses of pooled libraries indicated a significant shift due to CTC in both the lumen and the mucosa, with some OTUs unique to either treated or control ileum. DOTUR analysis revealed little overlap between control and treated communities at the 3% difference level, indicating unique ileal communities in the presence of CTC.

Enterotoxigenic Escherichia coli (ETEC) infections result in large economic losses in the swine industry worldwide. The organism causes diarrhea by adhering to and colonizing enterocytes in the small intestines. While much progress has been made in understanding the pathogenesis of ETEC, no homologous intestinal epithelial cultures suitable for studying porcine ETEC pathogenesis have been described prior to this report. In the current study, we investigated the adherence of various porcine ETEC strains to two porcine (IPEC-1 and IPEC-J2) and one human (INT-407) small intestinal epithelial cell lines. Each cell line was assessed for its ability to support the adherence of E. coli expressing fimbrial adhesins K88ab, K88ac, K88ad, K99, F41, 987P, and F18. Wild-type ETEC expressing K88ab, K88ac, and K88ad efficiently bound to both IPEC-1 and IPEC-J2 cells. An ETEC strain expressing both K99 and F41 bound heavily to both porcine cell lines but an E. coli strain expressing only K99 bound very poorly to these cells. E. coli expressing F18 adhesin strongly bound to IPEC-1 cells but did not adhere to IPEC-J2 cells. The E. coli strains G58-1 and 711 which express no fimbrial adhesins and those that express 987P fimbriae failed to bind to either porcine cell line. Only strains B41 and K12:K99 bound in abundance to INT-407 cells. The binding of porcine ETEC to IPEC-J2, IPEC-1 and INT-407 with varying affinities, together with lack of binding of 987P ETEC and non-fimbriated E. coli strains, suggests strain-specific E. coli binding to these cell lines. These findings suggest the potential usefulness of porcine intestinal cell lines for studying ETEC pathogenesis.

Two-photon absorption spectrum of aniline is recorded using thermal lens effect with optical parametric oscillator as the pump source. Studies show that the two-photon absorption spectrum observed corresponds to 1A1 → 1B2 transition of aniline.

It has been suggested that changes in diet at weaning in pigs induce intestinal inflammation which may be mediated through toll-like receptors (TLRs). We hypothesized that the use of antibiotics as growth promoters and subsequent changes in intestinal microbiota may mediate changes in the expression of TLRs in the intestine. Thus, this study was performed to assess the changes in intestinal TLRs in weanling pigs in response to the use of chlortetracycline as a growth promotant, and under gnotobiotic conditions. Eighteen cesarean-derived half-sib piglets were divided into three groups; antibiotic-fed, control (normal-fed) and gnotobiotic groups.TLR-2, -4, -5 and -9 gene expression and abundance of TLR-2 and -9 proteins in the ileum of pigs was assessed at 5 wk of age. No significant differences (p ?0.5) in TLR-2, -4, -5 and -9transcript levels and abundanceof TLR-2 and -9 proteins among three groups of pigswere observed.

A study was undertaken with the aim of reducing the concentration of cryoprotectants in surimi without adversely affecting frozen storage stability. Minced meat from a tropical fish, Nemipterus japonicus, was strained, water leached and mixed with different levels of sucrose-sorbitol (1:1) mixture (henceforth called sugar mixture), quick frozen at −35 °C and frozen stored at −20 °C. The surimi samples were subjected to storage stability studies for a period of 5 months. Water leaching resulted in slight absorption of water by meat and reduction in protein, fat and mineral contents. Surimi was found to have moderately white colour. Sensory evaluation studies were conducted on three products, viz., sausage, patty and cake, prepared using surimi containing different concentrations of sugar mixture. Sugar mixture content varying from 0% (control) to 4% in surimi resulted in products that were more acceptable to the taste panelists compared those with 6% and 8% sugar mixture. During frozen storage of surimi pH and total plate count remained nearly steady for all sugar mixture concentrations and throughout the storage period. Moisture content appeared to remain constant during storage, but decreased with increase in sugar concentration. The salt soluble nitrogen content of surimi and gel strength of sausage prepared from it decreased with storage period in all surimi samples, and increased with sugar mixture concentration. Expressible water content of surimi sausage showed an increasing trend with storage period of surimi and a decreasing trend with sugar mixture concentration. Sensory evaluation parameters—elasticity, sweetness and preference—remained more or less steady during storage. However elasticity and sweetness increased and preference decreased with sugar mixture concentration beyond 4%. Elasticity and gel strength of surimi sausage seemed to be much lower for control compared to even the lowest concentration of sugar (2%) used. A concentration of 2 to 4% sucrose-sorbitol mixture is well-accepted by the consumers in products—surimi sausage, patty and cake and at this range of concentration surimi could be well-preserved at −20 °C for at least 5 months.

The wavelength dependence of thermal lens signal from organic dyes are studied using dual beam thermal lens technique. It is found that the profile of thermal lens spectrum widely differ from the conventional absorption spectrum in the case of rhodamine B unlike in the case of crystal violet. This is explained on the basis of varying contribution of nonradiative relaxations from the excited vibronic levels.