Search results for: human skin and hair cells

Authors: Khaled M. Mohamed

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Background: P-Phenylenediamine (PPD) is used in the manufacture of hair dyes and skin decoration. In some developing countries, suicidal, homicidal and accidental cases by PPD were recorded. In this work, a sensitive LC-MS/MS method for determination of PPD and its metabolites N-acetyl-p-phenylenediamine (MAPPD) and N,N-diacetyl-p-phenylenediamine (DAPPD) in human urine has been developed and validated. Methods: PPD, MAPPD and DAPPD were extracted from urine by methylene chloride at alkaline pH. Acetanilide was used as internal standard (IS). The analytes and IS were separated on an Eclipse XDB- C18 column (150 X 4.6 mm, 5 µm) using a mobile phase of acetonitrile-1% formic acid in gradient elution. Detection was performed by LC-MS/MS using electrospray positive ionization under multiple reaction-monitoring mode. The transition ions m/z 109 → 92, m/z 151 → 92, m/z 193 → 92, and m/z 136 → 77 were selected for the quantification of PPD, MAPPD, DAPPD, and IS, respectively. Results: Calibration curves were linear in the range 10–2000 ng/mL for all analytes. The mean recoveries for PPD, MAPPD and DAPPD were 57.62, 74.19 and 50.99%, respectively. Intra-assay and inter-assay imprecisions were within 1.58–9.52% and 5.43–9.45% respectively for PPD, MAPPD and DAPPD. Inter-assay accuracies were within -7.43 and 7.36 for all compounds. PPD, MAPPD and DAPPD were stable in urine at –20 degrees for 24 hours. Conclusions: The method was successfully applied to the analysis of PPD, MAPPD and DAPPD in urine samples collected from suicidal cases.

Keywords: p-Phenylenediamine, metabolites, urine, LC-MS/MS, validation

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Authors: Asaf Toker

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Matricelf developed a technology that enables the production of autologous engineered tissue composed of matrix and cells derived from patients Omentum biopsy. The platform showed remarkable pre-clinical results for several medical conditions. The company recently licensed the technology that enabled scientist at Tel Aviv university that 3D printed a human heart from human cells and matrix for the first time in human history. The company plans to conduct its first human clinical trial for Acute Spinal Cord Injury (SCI) early in 2023.

Keywords: tissue engineering, regenerative medicine, spinal Cord Injury, autologous implants, iPSC

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Authors: Eric Chabrol, Sidonia B.G. Eckle, Renate de Boer, James McCluskey, Jamie Rossjohn, Mirjam H.M. Heemskerk, Stephanie Gras

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αβ and γδ T-cells are disparate T-cell lineages that, via their use of either αβ or γδ T-cell antigen receptors (TCRs) respectively, can respond to distinct antigens. Here we characterise a new population of human T-cells, term δβ T-cells, that express TCRs comprising a TCR-δ variable gene fused to a Joining-α/Constant-α domain, paired with an array of TCR-β chains. We characterised the cellular, functional, biophysical and structural characteristic feature of this new T-cells population that reveal some new insight into TCR diversity. We provide molecular bases of how δβ T-cells can recognise viral peptide presented by Human Leukocyte Antigen (HLA) molecule. Our findings highlight how components from αβ and γδTCR gene loci can recombine to confer antigen specificity thus expanding our understanding of T-cell biology and TCR diversity.

Keywords: new delta-beta TCR, HLA, viral peptide, structural immunology

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Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee

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In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.

Keywords: apoptosis, cancer, G1 arrest, panaxydol

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Authors: Sokol Isaraj, Lorela Bendo

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Scars resulting from surgery, injury, or burns have a physical and psychological impact on the affected patient. Although a number of treatments are available, nano fat grafting is an effective treatment for scars. Nano fat is a liquid suspension rich in stem cells obtained by mechanical emulsification. Nano fat grafting was performed in 10 cases to correct rhytides, surgical scars, and post-burn scars between January 2022 and April 2022. Fat was aspirated from the lower abdomen or trochanteric region. After emulsification and filtration protocol, the resulting nano fat liquid was injected intradermally and subdermally. All patients filled out a questionnaire at three months post-treatment, which consisted of questions regarding the grade of improvement of skin and recommendation of the procedure. The clinical results were apparent between 2 and 3 weeks after the treatment. All patients confirmed an improvement in skin texture and quality. The most significant improvement was seen in pigmentation and pliability. No complications were reported. Nano fat seems to be a safe and effective treatment in scar treatment and skin rejuvenation.

Keywords: fat grafting, fat transfer, micro fat, nano fat

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Authors: Sokol Isaraj, Lorela Bendo

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Scars resulting from surgery, injury, or burns have a physical and psychological impact on the affected patient. Although a number of treatments are available, nano fat grafting is an effective treatment for scars. Nano fat is a liquid suspension rich in stem cells obtained by mechanical emulsification. Nano fat grafting was performed in 10 cases to correct rhytides, surgical scars and post-burn scars between January 2022 and April 2022. Fat was aspirated from the lower abdomen or trochanteric region. After emulsification and filtration protocol, the resulting nano fat liquid was injected intradermally and subdermally. All patients filled out a questionnaire at 3 months post-treatment which consisted of questions regarding the grade of improvement of skin and recommendation of the procedure. The clinical results were apparent between 2 and 3 weeks after the treatment. All patients confirmed an improvement in skin texture and quality. The most significant improvement was seen in pigmentation and pliability. No complications were reported. Nano fat seems to be a safe and effective treatment for scar treatment and skin rejuvenation.

Keywords: fat grafting, fat transfer, microfat, nanofat

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Authors: Hasan Mahmud Reza

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Extensive studies of the human umbilical cord, both basic and translational, over the last three decades have unveiled a plethora of information. The cord lining harbors at least two phenotypically different multipotent stem cells: mesenchymal stem cells (MSCs) and cord lining epithelial stem cells (CLECs). These cells exhibit a mixed genetic profiling of both embryonic and adult stem cells, hence display a broader stem features than cells from other sources. We have observed that umbilical cord-derived cells are immunologically privileged and non-tumorigenic by animal study. These cells are ethically acceptable, thus provides a significant advantage over other stem cells. The high proliferative capacity, viability, differentiation potential, and superior harvest of these cells have made them better candidates in comparison to contemporary adult stem cells. Following 30 replication cycles, these cells have been observed to retain their stemness, with their phenotype and karyotype intact. Transplantation of bioengineered CLEC sheets in limbal stem cell-deficient rabbit eyes resulted in regeneration of clear cornea with phenotypic expression of the normal cornea-specific epithelial cytokeratin markers. The striking features of low immunogenicity protecting self along with co-transplanted allografts from rejection largely define the transplantation potential of umbilical cord-derived stem cells.

Keywords: cord lining epithelial stem cells, mesenchymal stem cell, regenerative medicine, umbilical cord

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Authors: Jan F. Talts, Amit Saxena, Kåre Engkilde

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Chemotherapy-induced peripheral neuropathy (CIPN) is one of the most frequent side effects caused by anti-neoplastic agents, with a prevalence from 19 % to 85 %. Clinically, CIPN is a mostly sensory neuropathy leading to pain and to motor and autonomic changes. Due to its high prevalence among cancer patients, CIPN constitutes a major problem for both cancer patients and survivors, especially because currently, there is no single effective method of preventing CIPN. Hearing loss is the most common form of sensory impairment in humans and can be caused by ototoxic chemical compounds such as chemotherapy (platinum-based antineoplastic agents).In rodents, single or repeated cisplatin injections induce peripheral neuropathy and hearing impairment mimicking human disorder, allowing studying the efficacy of new pharmacological candidates in chemotherapy-induced hearing loss and peripheral neuropathy. RNA sequencing data from full term amniotic fluid (TAF) mesenchymal stemcell (MSC) clones was used to identify neural-specific markers present on TAF-MSC. Several prospective neural markers were tested by flow cytometry on cultured TAF-MSC. One of these markers was used for cell-sorting using Tyto MACSQuant cell sorter, and the neural marker positive cell population was expanded for several passages to the final therapeutic product stage. Peripheral neuropathy and hearing loss was induced in mice by administration of cisplatin in three week-long cycles. The efficacy of neural-specific TAF-MSC in treating hearing loss and pain perception was evaluated by administration of three injections of 3 million cells/kg by intravenous route or three injections of 3 million cells/kg by intra-arterial route after each cisplatin cycle treatment. Auditory brainstem responses (ABR) are electric potentials recorded from scalp electrodes, and the first ABR wave represents the summed activity of the auditory nerve fibers contacting the inner hair cells. For ABR studies, mice were anesthetized, then earphones were placed in the left ear of each mouse, an active electrode was placed in the vertex of the skull, a reference electrode under the skin of the mastoid bone, and a ground electrode in the neck skin. The stimuli consisted of tone pips of five frequencies (2, 4, 6, 12, 16, and 24 kHz) at various sound levels (from 0 to 90 dB) ranging to cover the mouse auditory frequency range. The von Frey test was used to assess the onset and maintenance of mechanical allodynia over time. Mice were placed in clear plexiglass cages on an elevated mesh floor and tested after 30 min of habituation. Mechanical paw withdrawal threshold was examined using an electronic von Frey anesthesiometer. Cisplatin groups treated with three injections of 3 million cells/kg by intravenous route and three injections of 3 million cells/kg by intra-arterial route after each cisplatin cycle treatment presented, a significant increase of hearing acuity characterized by a decrease of ABR threshold and a decrease of neuropathic pain characterized by an increase of von Frey paw withdrawal threshold compared to controls only receiving cisplatin. This study shows that treatment with MSCselected for neural specificity presents significant positive efficacy on the chemotherapy-induced neuropathic pain and the chemotherapy-induced hearing loss.

Keywords: mesenchymal stem cell, peripheral neuropathy, amniotic fluid, regenerative medicine

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Authors: Wehandaka Pancapalaga

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This study aimed to analyze the water content, yield, fat content, protein content, viscosity, gel strength, pH, melting and organoleptic rabbit skin gelatin with acetic acid extraction levels are different. The materials used in this study were Rex rabbit skin male. Treatments that P1 = the extraction of acetic acid 2% (v / v); P2 = the extraction of acetic acid 3% (v / v); P3 = the extraction of acetic acid 4 % (v / v). P5 = the extraction of acetic acid 5% (v / v). The results showed that the greater the concentration of acetic acid as the extraction of rabbit skin can reduce the water content and fat content of rabbit skin gelatin but increase the protein content, viscosity, pH, gel strength, yield and melting point rabbit skin gelatin. texture, color and smell of gelatin rabbits there were no differences with cow skin gelatin. The results showed that the quality of rabbit skin gelatin accordance Indonesian National Standard (SNI). Conclusion 5% acetic acid extraction produces the best quality gelatin.

Keywords: gelatin, skin rabbit, acetic acid extraction, quality

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Authors: Azizollah Khodakaram- Tafti, Ali Mohammadi, Ghasem Farjanikish

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Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens of cattle worldwide caused by Pestivirus genus, Flaviviridae family.The aim of the present study was to comparison several diagnostic methods and determine the prevalence of BVDV infection for the first time in dairy herds of Fars province, Iran. For initial screening, a total of 400 blood samples were randomly collected from 12 industrial dairy herds and analyzed using reverse transcription (RT)-PCR on the buffy coat. In the second step, blood samples and also ear notch biopsies were collected from 100 cattle of infected farms and tested by antigen capture ELISA (ACE), RT-PCR and immunohistochemistry (IHC). The results of nested RT-PCR (outer primers 0I100/1400R and inner primers BD1/BD2) was successful in 16 out of 400 buffy coat samples (4%) as acute infection in initial screening. Also, 8 out of 100 samples (2%) were positive as persistent infection (PI) by all of the diagnostic tests similarly including RT-PCR, ACE and IHC on buffy coat, serum and skin samples, respectively. Immunoreactivity for bovine BVDV antigen as brown, coarsely to finely granular was observed within the cytoplasm of epithelial cells of epidermis and hair follicles and also subcutaneous stromal cells. These findings confirm the importance of monitoring BVDV infection in cattle of this region and suggest detection and elimination of PI calves for controlling and eradication of this disease.

Keywords: antigen capture ELISA, bovine viral diarrhea virus, immunohistochemistry, RT-PCR, cattle

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Authors: Wassim Guermazi, Saoussan Boukhris, Neila Annabi Trabelsi, Tarek Rebai, Alya Sellami-Kamoun, Habib Ayadi

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This work investigates the protective effects of the microalga Halamphora sp. extract (H. Ext) as a natural product on lead-intoxicated liver and kidney human cells in vitro and in vivo on rats wistar. HepG2 cells line derived from human hepatocellular carcinoma and HEK293 cells line derived from human embryonic kidney were used for the in vitro study. The analysis of the fatty acids methyl esters of the extract was performed by a GC/MS. Four groups of rats, each of which was composed of six animals, were used for the in vivo experiment. The pretreatment of HepG2 and HEK293 cells line with the extract (100 µg mL-1) significantly (p < 0.05) protected against cytotoxicity induced by lead exposure. In vivo, the biochemical parameters in serum, namely malondialdehyde level (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, were measured in supernatants of organ homogenates. H. Ext was found to be rich in fatty acids, essentially palmitic and palmitoleic accounting respectively 29.46% and 42.07% of total fatty acids. Both in vitro and in vivo, the co-treatment with H. Ext allowed the protection of the liver and kidney cells structure, as well as the significant preservation of normal antioxidant and biochemical parameters in rats. Halamphora extract rich in fatty acids has been proven to be effective in protection against Pb-induced toxicity.

Keywords: microalga extract, human cells line, fatty acid, lead exposure, oxidative stress, rats

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Authors: Xian-Peng Jiang, Catherine C. Baucom, Toby Jiang, Robert L. Elliott

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HLA-G binds to the inhibitory receptors of uterine NK cells and plays an important role in protection of fetal cells from maternal NK lysis. HLA-G also mediates tumor escape, but the immunosuppressive role is often neglected. These studies have focused on the examination of HLA-G expression in human breast carcinoma and HLA-G immunosuppressive role in NK cytolysis. We examined HLA-G expression in breast cell lines by real time PCR, ELISA and immunofluorescent staining. We treated the breast cancer cell lines with anti-human HLA-G antibody or progesterone. Then, NK cytolysis was measured by using MTT assay. We find that breast carcinoma cell lines increase the expression of HLA-G mRNA and protein, compared to normal cells. Blocking HLA-G of the breast cancer cells by the antibody increases NK cytolysis. Progesterone upregulates HLA-G mRNA and protein of human breast cancer cell lines. The increased HLA-G expression suppresses NK cytolysis. In summary, human breast carcinoma overexpress HLA-G immunosuppressive molecules. Blocking HLA-G protein by antibody improves NK cytolysis. In contrast, upregulation of HLA-G expression by progesterone impairs NK cytolytic function. Thus, HLA-G is a new immunosuppressive checkpoint and potential cancer immunotherapeutic target.

Keywords: HLA-G, Breast carcinoma, NK cells, Immunosuppressive checkpoint

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Authors: M. J. Kang, Y. N. Jo, H. H. Yoo

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Pendulum tests were used to identify a stretch reflex and diagnose spasticity. Some researches tried to make a mathematical model to simulate the motions. Thighs are subject to compressive forces due to gravity during a pendulum test. Therefore, it affects knee trajectories. However, the most studies on the pendulum tests did not consider that conditions. We used Kelvin-Voight model as compression model of skin and muscles. In this study, we investigated viscoelastic behaviors of skin and muscles using gelatin blocks from experiments of the vibration of the compliantly supported beam. Then we calculated a dynamic stiffness and loss factors from the experiment and estimated a damping coefficient of the model. We also did pendulum tests of human lower limbs to validate the stiffness and damping coefficient of a skin model. To simulate the pendulum motion, we derive equations of motion. We used stretch reflex activation model to estimate muscle forces induced by the stretch reflex. To validate the results, we compared the activation with electromyography signals during experiments. The compression behavior of skin and muscles in this study can be applied to analyze sitting posture as wee as developing surgical techniques.

Keywords: Kelvin-Voight model, pendulum test, skin and muscles under compression, stretch reflex

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Authors: Zhaoguo Liu, Cunsi Shen, Yin Lu

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Growing evidence has shown that menthol has potent anticancer activity in various human cancers. However, its effect on skin cancer remains largely unknown. In the present study, we investigated the chemopreventive potential of menthol against 7, 12-dimethylbenz[a] anthracene(DMBA)/12-O-tetradecanoylphorbol 13-acetate (TPA)-induced skin tumorigenesis in ICR mice. Our results showed that menthol significantly inhibited TPA-induced inflammatory responses and pro-inflammatory cytokine release. We also found that menthol treatment significantly inhibited TPA-induced lipid peroxidation (LPO), mouse UDP-glucumno-syltransferase (UGT), mouse NADH Dehydrogenase, Quinone 1 (NQO1) release. Furthermore, we found menthol treatment significantly inhibited the tumor incidence and number of tumors (P < 0.001). Interestingly, we observed that menthol treatment significantly inhibited TPA-induced altered activity of NF-κB in skin tumor. Consistently, menthol-treated tumors also showed significantly suppressed the Ras-Raf-ERK signaling pathway. Thus, our results suggest that menthol inhibits DMBA/TPA-induced skin tumorigenesis by attenuating the Ras and inhibiting NF-κB activity via inhibition of inflammation responses and pro-inflammatory cytokine release.

Keywords: DMBA/TPA, NF-κB, Ras-Raf-ERK, skin tumorigenesis

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Authors: Dwarika P. Shrestha, Dipendra Gurung, Rushma Shrestha, Inger Rosdahl

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Introduction: Skin diseases are one of the most common health problems in Nepal. The objectives of this study are to determine the prevalence of skin diseases and impact on quality of life in rural areas in Nepal. Materials and methods: A house-to-house survey was conducted, to obtain socio-demographic data and identify individuals with skin diseases, followed by health camps, where the villagers were examined. A pilot study was conducted in one village, which was then extended to 10 villages in 4 districts. To assess the impact on quality of life, the villagers were interviewed with Skin Disease Disability Index. This is a questionnaire developed and validated by the authors for use in Nepal. Results: In the pilot study, the overall prevalence of skin diseases was 20.1% (645/3207). In the additional 10 villages with 7348 (3651/3787 m/f) inhabitants, 1862 (721/1141 m/f, mean age 31.4 years) had one or more skin diseases. The overall prevalence of skin diseases was 25%. The most common skin disease categories were eczemas (13.7%, percentage among all inhabitants) pigment disorders (6.8%), fungal infections (4.9%), nevi (3.7%) and urticaria (2.9%). These five most common skin disease categories comprise 71% of all skin diseases seen in the study. The mean skin disease disability index score was 13.7, indicating very large impact on the quality of life. Conclusions: This population-based study shows that skin diseases are very common in the rural areas of Nepal and have significant impact on quality of life. Targeted intervention at the primary health care level should help to reduce the health burden due to skin diseases.

Keywords: prevalence and pattern of skin diseases, impact on quality of life, rural Nepal, interventions

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Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

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Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

Procedia PDF Downloads 191

Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

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Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

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Authors: D. R. Apoorva

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Ichthyosis follicularis, alopecia, and photophobia (IFAP) syndrome is a rare oculocutaneous disorder of genetic origin. This disorder results from mutations in the membrane-bound transcription factor protease site, two genes that impair cholesterol homeostasis, and the ability to cope with endoplasmic reticulum stress. We report a rare case of IFAP syndrome with an atypical presentation, and it was interesting to note that the child had patchy non-scarring alopecia over the scalp along with unilateral madarosis. To our best knowledge, this unique presentation has not been described earlier. The child presented with photophobia and unilateral ptosis. The child also had short stature and intellectual disability. Skin histopathology was nonspecific and consisted of dilated hair follicles with keratin plugs extending above the skin surface. This rare oculocutaneous disorder requires proper documentation so that identification of its variants may be possible in the future. Early recognition of atypical presentations can help in preventing cardiovascular complications, which remain the major cause of death.

Keywords: alopecia, photophobia, ichthyosis follicularis, IFAP syndrome

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Authors: S. P. Gomes, D. C. Goncalves, E. Ribeiro, M. C. L. Seelaender

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Skin is susceptible to photo damage induced by exposure to sunlight, or ultraviolet (UV) radiation, which induces breakdown of extracellular matrix, DNA degradation, skin cell lesion and apoptosis, and development of cancer. Phytonutrients demonstrate protective effects against UV damage. The purpose of this study was evaluating the effect of an antioxidant juice (AJ) contaning Brazilian natural products upon skin damage. The juice was produced by Metabolics®. Male Wistar rats were divided in 4 groups: Animals receiving the antioxidant juice (AJ): orange, carrot, honey, tomato extract, avocado, ginger and camu-camu (Brazilian fruit, a major source of vitamin C) ad libitum for 21 days; or water (C), subdivided in groups exposed or not to UV radiation for 2 non consecutive days, during five hours each day, after 15 days of juice supplementation. On the 22nd day, rats were killed by decapitation and epithelium samples from the dorsal skin removed, fixed in bouin and embedded in paraffin. The sections were stained with hematoxylin and eosin or mallory and picrosirius red. Isolated DNA was submitted to electrophoresis (1.8% agarose gel, 0.5% ethidium bromide). UV radiation significantly induced sunburn of superficial epithelial cells of C, AJ treatment reduced this effect. Collagen changes were observed in UV groups, yet AJ treatment prevented collagen degradation. UV radiation induced significant DNA degradation, in C, which was prevented by AJ treatment. The antioxidant juice consumed chronically protected against acute skin damage.

Keywords: nutraceuticals, antioxidants, photoprotection, uv radiation

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Authors: Malay K. Das, Bhupen Kalita

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The poor oral bioavailability of resveratrol (RSV) due to presystemic metabolism can be avoided via dermal route of administration. The hydrophilic-lipophilic nature of resveratrol-phospholipid complex (RSVPs) favors the delivery of resveratrol via the skin. The RSVPs embedded polymeric patch with moderate adhesiveness was developed for dermal application for sustained anti-inflammatory effect. The prepared patches were evaluated for various physicochemical properties, surface morphology by SEM, TEM, and compatibility of patch components by FT-IR and DSC studies. The dermal flux of the optimized patch formulation was found to be at 4.28 ± 0.48 mg/cm2/24 h. The analysis of skin extract after permeation study revealed the presence of resveratrol, which confirmed the localization of RSVPs in the skin. The stability of RSVPs in the polymeric patch and the physiologic environment was confirmed by FE-SEM studies on the patches after drug release and skin permeation studies. The RSVPs particles released from the polymer matrix maintaining the structural integrity and permeate the keratinized horney layer of skin. The optimized patch formulation showed sustained anti-inflammatory effect (84.10% inhibition of inflammation at 24 h) in carrageenan-induced rat paw edema model compared to marketed diclofenac sodium gel (39.58% inhibition of inflammation at 24 h). The CLSM study confirmed the localization of RSVPs for a longer period, thus enabling drug targeting to the dermis for sustained anti-inflammatory effect. Histological studies with phase contrast trinocular microscope suggested no alteration of skin integrity and no evidence of the presence of inflammatory cells after exposure to the permeants. The patch was found to be safe for skin application as evaluated by Draize method for skin irritation scoring in a rabbit model. These results suggest the therapeutic efficacy of the developed patch in both acute and chronic inflammatory diseases.

Keywords: resveratrol-phospholipid complex, skin delivery, sustained anti-inflammatory effect, inflammatory diseases, dermal patch

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Authors: Sheikha Turki Alketbi

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Objectives: 1. Documenting the spontaneous resolution of AML following the initiation of anti-TB therapy. 2. Presenting an uncommon type of relapse in Acute Myeloid Leukemia. 3. Highlighting the role of immune markers in the diagnosis of Leukemia cutis. 4. Exploring and highlighting the possibility of skin relapse as the exclusive manifestation, even when skin involvement is known secondary manifestation in AML. Background: Spontaneous remission of Acute Myeloid Leukemia (AML) is a rare phenomenon that has only been reported in some case reports, usually following severe infections. Some studies have described the occurrence of tuberculosis (TB) infection with AML, usually after starting chemotherapy. Spontaneous resolution of AML after starting anti TB therapy (ATT), without starting chemotherapy has never been described in the literature. Moreover, Leukemia cutis is another rare skin manifestation of Acute Myeloid Leukemia as a result of infiltration of the skin or subcutaneous tissue by leukemic cells, in which can present during, precedes, after or independently of systemic leukemia. Methods: Here, we present a case of a 13-year-old male who presented with fever, weight loss, lethargy, epistaxis, bruising and dry cough and was later diagnosed with AML. Before initiating leukemia treatment, the patient was tested for TB and was found to have active TB infection. His leukemia treatment was postponed to clear the TB infection and he was commenced on ATT. Two months later, repeat blood film and bone marrow biopsy showed resolution of his AML. The patient remained in remission for 1 month, after which he presented with symmetrical blue purple well-defined round indurated plaques on the chest and thighs. Our differentials were leukemia cutis and Kaposi sarcoma. Results: Skin Biopsy with immune markers done, showed a picture of Acute Myeloid Leukemia. Immunohistochemistry (IHC) showed neoplastic cells diffusely and strongly positive for LCA, CD2, CD31, MPO, CD117, Lysozymes and TDT, and moderately positive for CD34, CD99, CD43 and CD6 And patchy for CD68. Ki67 showed 60% proliferation index. They were negative for the remaining markers. This suggested acute myeloid leukemia (AML). Conclusion: In summary, we present a rare case of TB with AML that resolved after treatment of TB with ATT but relapsed later as leukemia cutis. While skin involvement might occur as a secondary manifestation of AML, Skin relapse could be the only one.

Keywords: Leukemia cutis, Leukemia relapse, Acute Myeloid Leukemia, spontaneous resolution of AML

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Authors: Abujnah Dukali

Abstract:

Natural honey was assessed in cell culture system for its anticancer activity. Human leukemic cell line HL 60 was treated with honey and cultured for 5 days and cytotoxicity was calculated by MTT assay. Honey showed cytotoxicity with CC50 value of 174.20 µg/ml. Radical modulation activities was assessed by lipid peroxidation assay using egg lecithin. Honey showed antioxidant activity with EC50 value of 159.73 µg/ml. In addition, treatment with HL60 cells also resulted in nuclear DNA fragmentation, as seen in agarose gel electrophoresis. This is a hallmark of cells undergoing apoptosis. Confirmation of apoptosis was performed by staining the cells with Annexin V and FACS analysis. Apoptosis is an active, genetically regulated disassembly of the cell form within. Disassembly creates changes in the phospholipid content of the cytoplasmic membrane outer leaflet. Phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. The human anticoagulant, annexin V, is a Ca2+-dependent phospholipid protein with a high affinity for PS. Annexin V labeled with fluorescein can identify apoptotic cells in the population It is a confirmatory test for apoptosis. Annexin V-positive cells were defined as apoptotic cells. Since honey shows both antioxidant activity and cytotoxicity at almost the same concentration, it can prevent the free radical induced cancer as prophylactic agent and kill the cancer cells by apoptotic process as a chemotherapeutic agent. Everyday intake of honey can prevent the cancer induction.

Keywords: anticancer, cells, DNA, honey

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Authors: Ayse Ozge Demir, Nihat Mert

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This study has been carried out in order to determine the polymorphic traits of various biochemical parameters in goat breeds which are native to Turkey. For this purpose, Angora and Hair goats breeds were chosen as live materials. Two different herds for each breed were selected from Ankara and Antalya, respectively. Blood samples were taken from a total of 120 goats aged between 2 and 4 which was made up of 60 Angora goats and 60 Hair goats. All which derived equally from 4 lots of herds. Analyses were performed for the polymorphic determination of the Haemoglobin (Hb), Transferrine (Tf), Ceruloplasmin (Cp) and Glutathione (GSH). Hb types were determined by starch gel electrophoresis and Tf types were detected by SDS-PAGE electrophoresis. Furthermore, Cp and GSH analyses were performed by spectrophotometrically. Following the analysis, Hb types were found as 3 genotypes (AA, AB, BB) controlled by 2 allel genes. Tf types were found as 6 genotypes (AA, AB, AC, BB, BC, CC) controlled by 3 allele genes. Findings for Hb was in line with the Hardy-Weinberg Equilibrium (HWE) in Angora goats while the Hair goat was not found to be in line. Moreover, Tf was found in line with the HWE for 2 separate goat breeds. The levels of Cp and GSH of two breeds were significantly different from other (P<0.0001). The findings are recorded as a source of reference for prospective polymorphism studies.

Keywords: electrophoresis, genetic resources, goats, spectrophotometer

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Authors: Beata Kaczmarek-Szczepańska, Lidia Zasada

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Marine collagens such as fish skin, bone, cartilage, and scales, including both marine vertebrates and invertebrates sources, are more bioavailable compared to bovine or porcine collagen and have a higher absorption capability and more rapid bloodstream circulation due to their low molecular weight and small particle size. Fish skin may be used as a source of bioactive compounds. The advantage is that fish skin is a by-product of the food industry. The subject of the study is a lyophilizate consisting of a mixture of compounds from the group of glycosaminoglycans and collagen obtained as a result of processing fish skins. Bioactive compounds may find biomedical use in the production of dressing materials for wounds or in tissue engineering for the production of scaffolds for cells. Collagen and glycosaminoglycans were isolated from Salmo salar skin. The final mixture was obtained by the freeze-drying method. In the obtained lyophilizate, the content of amino acids was studied as well as the presence of polysaccharides. The studies showed the presence of glycine, proline, and hydroxyproline, which are the main amino acids of collagen. The HPLC analysis showed the presence of glucose which is a product of polysaccharides hydrolyzation and then reduction of glucuronic acid. It may be assumed that the lyophilizate contains both collagen and polysaccharide, which is probably hyaluronic acid. Acknowledgment: This work was carried out as a result of research project no. TANGO-V-A/0020/2021 financed by the National Centre for Research and Development.

Keywords: collagen, glycosaminoglycans, bioactive compounds, fish skin

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Authors: Mohamed Z. Sayed-Ahmed, Ahmed M. Ahdy, Emad E. Younis, Sabry A. El-Khodary

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Little research information is available on the prevalence of diseases of donkeys in Egypt. Across sectional study was undertaken between March 2009 and February 2010 to verify the prevalence of skin affection of donkeys. A total of 1134 donkeys in northern Egypt were investigated. A questionnaire was constructed to verify the number of infected contact animals as well as the associated factors. Physical examination was carried out, and the distribution of skin lesions was recorded. Skin scraping and biopsy were obtained to perform bacteriological, mycological, and histopathological examinations. Thirty-five (3.09%) out of 1134 noticed donkeys had skin affections including mange (18/35), dermatophytosis (6/35), bacterial dermatitis (6/35) urticaria (2/35) and allergic dermatitis (3/35). The present results indicate that mange and dermatophytosis are the prevalent skin diseases in donkeys. Contact with other animal species of contaminated environment may contribute to the occurrence of the diseases.

Keywords: donkeys, Egypt, prevalence, skin affection

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Authors: Kirti, Samanta Sekhar Khora

Abstract:

Inflation serves different purposes in different organisms and adds beauty to their behavioral attributes. Pufferfishes are also known as blowfish, swellfish, and globefish due to their remarkable ability to puff themselves up like a balloon when threatened. This ability to inflate can be correlated with anatomical features that are unique to pufferfishes. Pufferfish skin provides a rigid framework to support the body contents and a flexible covering to allow whatever changes are necessary for remarkable inflation mechanism. Skin, the outer covering of animals is made up of collagen fibers arranged in more or less ordered arrays. The ventral skin of pufferfish stretches more than dorsal skin during inflation. So, this study is of much of the interest in comparing the structure and mechanical properties of these two skin regions. The collagen fibers were found to be arranged in different ordered arrays for ventral and dorsal skin and concentration of fibers were also found to be different for these two skin parts. Scanning electron microscopy studies of the ventral skin showed a unidirectional arrangement of the collagen fibers, which provide more stretching capacity. Dorsal skin, on the other hand, has an orthogonal arrangement of fibers. This provides more stiffness to the ventral skin at the time of inflation. In this study, the possible role of collagen fibers was determined which significantly contributed to the remarkable inflation mechanism of pufferfishes.

Keywords: collagen, histology, inflation, pufferfish, scanning electron microscopy, Small-Angle X-Ray Scattering (SAXS), transmission electron microscopy

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Authors: Byeal-I Han, Michael Lee

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Ageing is associated with an increased risk of diseases such as cancer, and neurodegenerative disorders. Increased autophagy delays ageing and extends longevity. In this study, we investigated the role of autophagy in longevity using human foreskin fibroblast HS68 cells, in which a senescence-like growth arrest can be induced. In particular, cellular senescence is manifested by the irreversible cell cycle arrest, and may contribute to the ageing of organisms. The senescence state was measured with staining for senescence-associated β-galactosidase (SA-β-gal) activity that represents a sensitive and reliable marker to quantify senescent cells. We detected a significantly increased percentage (%) of SA-β-gal positive cells in HS68 cultures at passage 40 (63%) when compared with younger ones at passage 15 (0.5%). As expected, HS68 cells at passage 40 exhibited much lower proliferation rate than cells at passage 15. The basal levels of LC3 were measured by immunoblotting showing a comparison of LC3-I and LC3-II levels at 3 age-points in serially passaged HS68 cells. LC3-II/LC3-I ratio at different passage levels relative to β-actin levels of each band confirmed that cells at passage 34 showed lower conversion of non-autophagic LC3-I to autophagic LC3-II than the cells at passage 16. Furthermore, Cyto-ID autophagy assay also revealed that late passage cells showed lower autophagy than the early passage cells. Together, our findings suggest that senescence induction might be associated with decreased autophagy.

Keywords: ageing, autophagy, senescence, HS68

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Authors: Hirowati Ali, Aisyah Ellyanti, Dewi Rusnita, Septelia Inawati Wanandi

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Stem cell has been known for its potency to be differentiated in many cells. Recently stem cell has been used for many treatment of degenerative medicine. It is still controversy whether stem cell can be used for therapy or these cells can activate cancer stem cell. SCUBE2 is a novel secreted and membrane-anchored protein which has been reported to its role in better prognosis and inhibition of cancer cell proliferation. Our study aims to observe whether stem cell can up-regulate SCUBE2 gene in MCF7 breast cancer cell line. We used in vitro study using MCF-7 cell treated with stem cell derived from placenta Wharton's jelly which has been known for its stemness and widely used. Our results showed that MCF-7 cell line grows up rapidly in 6-well culture dish. Stem cell was cultured in 6-well dish. After 50%-60% MCF-7 confluence, we co-cultured these cells with stem cells for 24 hours and 48 hours. We hypothesize SCUBE2 gene which is previously known for its higher expression in better prognosis of breast cancer, is up-regulated after stem cells addition in MCF7 culture dishes.

Keywords: breast cancer cells, inhibition of cancer cells, mesenchymal stem cells, SCUBE2

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Authors: Shaira L. Kee, Michael Aaron G. Sy, Myles Joshua T. Tan, Hezerul Abdul Karim, Nouar AlDahoul

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Skin diseases are considered the fourth most common disease, with melanoma and non-melanoma skin cancer as the most common type of cancer in Caucasians. The alarming increase in Skin Cancer cases shows an urgent need for further research to improve diagnostic methods, as early diagnosis can significantly improve the 5-year survival rate. Machine Learning algorithms for image pattern analysis in diagnosing skin lesions can dramatically increase the accuracy rate of detection and decrease possible human errors. Several studies have shown the diagnostic performance of computer algorithms outperformed dermatologists. However, existing methods still need improvements to reduce diagnostic errors and generate efficient and accurate results. Our paper proposes an ensemble method to classify dermoscopic images into benign and malignant skin lesions. The experiments were conducted using the International Skin Imaging Collaboration (ISIC) image samples. The dataset contains 3,297 dermoscopic images with benign and malignant categories. The results show improvement in performance with an accuracy of 88% and an F1 score of 87%, outperforming other existing models such as support vector machine (SVM), Residual network (ResNet50), EfficientNetB0, EfficientNetB4, and VGG16.

Keywords: deep learning - VGG16 - efficientNet - CNN – ensemble – dermoscopic images - melanoma

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Authors: P. Renjitha, P. Hari Prakash

Abstract:

Auditory neuropathy spectrum disorder (ANSD) is an auditory disorder with normal cochlear outer hair cell function and disrupted auditory nerve function. It results in unique clinical characteristic with absent auditory brainstem response (ABR), absent acoustic reflex and the presence of otoacoustic emissions (OAE) and cochlear microphonics. The lesion site could be at cochlear inner hair cells, the synapse between the inner hair cells and type I auditory nerve fibers, and/or the auditory nerve itself. But the literatures on synchrony at higher auditory system are sporadic and are less understood. It might be interesting to see if there is a recovery of neural synchrony at higher auditory centers. Also, does the level at which the auditory system recovers with adequate synchrony to the extent of observable evoke response potentials (ERPs) can predict speech perception? In the current study, eight ANSD participants and healthy controls underwent detailed audiological assessment including ABR, auditory middle latency response (AMLR), and auditory late latency response (ALLR). AMLR was recorded for clicks and ALLR was evoked using 500Hz and 2 kHz tone bursts. Analysis revealed that the participant could be categorized into three groups. Group I (2/8) where ALLR was present only for 2kHz tone burst. Group II (4/8), where AMLR was absent and ALLR was seen for both the stimuli. Group III (2/8) consisted individuals with identifiable AMLR and ALLR for all the stimuli. The highest speech identification sore observed in ANSD group was 30% and hence considered having poor speech perception. Overall test result indicates that the site of neural synchrony recovery could be varying across individuals with ANSD. Some individuals show recovery of neural synchrony at the thalamocortical level while others show the same only at the cortical level. Within ALLR itself there could be variation across stimuli again could be related to neural synchrony. Nevertheless, none of these patterns could possible explain the speech perception ability of the individuals. Hence, it could be concluded that neural synchrony as measured by evoked potentials could not be a good clinical predictor speech perception.

Keywords: auditory late latency response, auditory middle latency response, auditory neuropathy spectrum disorder, correlation with speech identification score

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