Search results for: HUVEC cells

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

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Authors: Sandeep Kaur

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Phycoerythrin (PE) from the mesophilic filamentous cyanobacterium Nostoc piscinale PUPCCC 405.17, a good producer of phycobiliproteins, has been characterized in terms of their unit assembly and stability. The phycoerythrin was extracted by freeze-thawing the cells in water, concentrated by ammonium sulphate fractionation and purified by anion exchange chromatography. The purification process resulted in 2.90 fold increase in phycoerythrin purity reaching to 1.54. Sodium Dodecyl Sulphate- Polyacrylamide Gel Electrophoresis of purified PE demonstrated three protein bands of 14.3, 27.54 and 39.81 kDa. The native PE also showed one band of 125.87 kDa, assumed to be a dimer (αβ)2γ based on results of non-denaturing PAGE. Lyophilized powder PE was more stable compared to phycoerythrin in the solution. The half-life of dry PE is 80 days when stored at 4 °C under dark. The phycoerythrin from this organism has potential applications in food as natural colour and as a fluorescent marker.

Keywords: characterization, Nostoc piscinale, phycoerythrin, purification

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Authors: Carolina Schencke, Cristian Sandoval, Belgica Vasquez, Mariano Del Sol

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Introduction: In a burn injury, the successful repair requires not only the participation of various cells, such as granulocytes and fibroblasts, but also of collagen, which plays a crucial role as a structural and regulatory molecule of scar tissue. Since honey and ascorbic acid have presented a great therapeutic potential to cellular and structural level, experimental studies have proposed its combination in the treatment of wounds. Aim: To evaluate stereological and morphometric parameters of healing wounds, caused by burns, treated with honey Ulmo (Eucryphia cordifolia) unsupplemented, comparing its effect with Ulmo honey supplemented with ascorbic acid. Materials and Methods: Fifteen healthy adult guinea pigs (Cavia porcellus) were used, of both sexes, average weight 450 g from the Centro de Excelencia en Estudios Morfológicos y Quirúrgicos (CEMyQ) at the Universidad de La Frontera, Chile. The animals were divided at random into three groups: positive control (C+), honey only (H) and supplemented honey (SH) and were fed on pellets supplemented with ascorbic acid and water ad libitum, under ambient conditions controlled for temperature, ambient noise and a cycle of 12h light–darkness. The protocol for the experiment was approved by the Scientific Ethics Committee of the Universidad de La Frontera, Chile. The parameters measured were number density per area (NA), volume density (VV), and surface density (SV) of fibroblast; NA and VV of polymorphonuclear cells (PMN) and, evaluation of the content of collagen fibers in the scar dermis. One-way ANOVA was used for statistics analysis and its respective Post hoc tests. Results: The ANOVA analysis for NA, VV and SV of fibroblasts, NA and VV of PMN, and evaluation of collagen content, type I and III, showed that at least one group differs from other (P≤ 0.001). There were differences (P= 0.000) in NA of fibroblast between the groups [C+= 3599.560 mm-2 (SD= 764.461), H= 3355.336 mm-2 (SD= 699.443) and SH= 4253.025 mm-2 (SD= 1041.751)]. The VV and SV of fibroblast increased (P= 0.000) in the SH group [20.400% (SD= 5.897) and 100.876 mm2/mm3 (SD= 29.431), respectively], compared to the C+ [16.324% (SD= 7.719) and 81.676 mm2/mm3 (SD= 28.884), respectively). The mean values of NA and VV of PMN were higher (P= 0.000) in the H [756.875 mm-2 (SD= 516.489) and 2.686% (SD= 2.380), respectively) group. Regarding to the evaluation of the content of collagen fibers, type I and III, the one-way analysis of ANOVA showed a statistically significant difference (P< 0.05). The content of collagen fibers type I was higher in C+ (1988.292 μm2; SD= 1312.379), while the content of collagen fibers type III was higher in SH (1967.163 μm2; SD= 1047.944 μm2) group. Conclusions: The stereological results were correlated with the stage of healing observed for each group. These results suggest that the combination of honey with ascorbic acid potentiate the healing effect, where both participated synergistically.

Keywords: ascorbic acid, morphometry, stereology, Ulmo honey

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Authors: Vijayakameswara Rao Neralla, Jueun Jeon, Jae Hyung Park

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To develop a potential nanocarrier for diagnosis and treatment of rheumatoid arthritis (RA), we prepared a hyaluronic acid (HA)-5β-cholanic acid (CA) conjugate with an acid-labile ketal linker. This conjugate could self-assemble in aqueous conditions to produce pH-responsive HA-CA nanoparticles as potential carriers of the anti-inflammatory drug methotrexate (MTX). MTX was rapidly released from nanoparticles under inflamed synovial tissue in RA. In vitro cytotoxicity data showed that pH-responsive HA-CA nanoparticles were non-toxic to RAW 264.7 cells. In vivo biodistribution results confirmed that, after their systemic administration, pH-responsive HA-CA nanoparticles selectively accumulated in the inflamed joints of collagen-induced arthritis mice. These results indicate that pH-responsive HA-CA nanoparticles represent a promising candidate as a drug carrier for RA therapy.

Keywords: rheumatoid arthritis, hyaluronic acid, nanocarrier, self-assembly, MTX

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Authors: A. Mathieu, B. Aubry, E. Chhim, M. Jobe, M. Arnaud

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Present project consists in a study and a development of piezoelectric devices for supplying power to new generation pacemakers. They are miniaturized leadless implants without battery placed directly in right ventricle. Amongst different acceptable energy sources in cardiac environment, we choose the solution of a device based on conversion of the energy produced by pressure variation inside the heart into electrical energy. The proposed energy harvesters can meet the power requirements of pacemakers, and can be a good solution to solve the problem of regular surgical operation. With further development, proposed device should provide enough energy to allow pacemakers autonomy, and could be good candidate for next pacemaker generation.

Keywords: energy harvester, heart, leadless pacemaker, piezoelectric cells, pressure variation

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Authors: Feng Guo

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Human brain organoids, 3D brain tissue cultures derived from human pluripotent stem cells, hold promising potential in modeling neuroinflammation for a variety of neurological diseases. However, challenges remain in generating standardized human brain organoids that can recapitulate key physiological features of a human brain. Here, this study presents a series of organoids-on-a-chip systems to generate better human brain organoids and model neuroinflammation. By employing 3D printing and microfluidic 3D cell culture technologies, the study’s systems enable the reliable, scalable, and reproducible generation of human brain organoids. Compared with conventional protocols, this study’s method increased neural progenitor proliferation and reduced heterogeneity of human brain organoids. As a proof-of-concept application, the study applied this method to model substance use disorders.

Keywords: human brain organoids, microfluidics, organ-on-a-chip, neuroinflammation

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Authors: Carmela Nardelli, Laura Iaffaldano, Valentina Capobianco, Antonietta Tafuto, Maddalena Ferrigno, Angela Capone, Giuseppe Maria Maruotti, Maddalena Raia, Rosa Di Noto, Luigi Del Vecchio, Pasquale Martinelli, Lucio Pastore, Lucia Sacchetti

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Maternal obesity and nutrient excess in utero increase the risk of future metabolic diseases in the adult life. The mechanisms underlying this process are probably based on genetic, epigenetic alterations and changes in foetal nutrient supply. In mammals, the placenta is the main interface between foetus and mother, it regulates intrauterine development, modulates adaptive responses to sub optimal in uterus conditions and it is also an important source of human amniotic mesenchymal stem cells (hA-MSCs). We previously highlighted a specific microRNA (miRNA) profiling in amnion from obese (Ob) pregnant women, here we compared the miRNA expression profile of hA-MSCs isolated from (Ob) and control (Co) women, aimed to search for any alterations in metabolic pathways that could predispose the new-born to the obese phenotype. Methods: We isolated, at delivery, hA-MSCs from amnion of 16 Ob- and 7 Co-women with pre-pregnancy body mass index (mean/SEM) 40.3/1.8 and 22.4/1.0 kg/m2, respectively. hA-MSCs were phenotyped by flow cytometry. Globally, 384 miRNAs were evaluated by the TaqMan Array Human MicroRNA Panel v 1.0 (Applied Biosystems). By the TargetScan program we selected the target genes of the miRNAs differently expressed in Ob- vs Co-hA-MSCs; further, by KEGG database, we selected the statistical significant biological pathways. Results: The immunophenotype characterization confirmed the mesenchymal origin of the isolated hA-MSCs. A large percentage of the tested miRNAs, about 61.4% (232/378), was expressed in hA-MSCs, whereas 38.6% (146/378) was not. Most of the expressed miRNAs (89.2%, 207/232) did not differ between Ob- and Co-hA-MSCs and were not further investigated. Conversely, 4.8% of miRNAs (11/232) was higher and 6.0% (14/232) was lower in Ob- vs Co-hA-MSCs. Interestingly, 7/232 miRNAs were obesity-specific, being expressed only in hA-MSCs isolated from obese women. Bioinformatics showed that these miRNAs significantly regulated (P<0.001) genes belonging to several metabolic pathways, i.e. MAPK signalling, actin cytoskeleton, focal adhesion, axon guidance, insulin signaling, etc. Conclusions: Our preliminary data highlight an altered miRNA profile in Ob- vs Co-hA-MSCs and suggest that an epigenetic miRNA-based mechanism of gene regulation could affect pathways involved in placental growth and function, thereby potentially increasing the newborn’s risk of metabolic diseases in the adult life.

Keywords: hA-MSCs, obesity, miRNA, biosystem

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Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

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Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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Authors: Shin-Yi Mao, Jiashing Yu

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Decellularized adipose tissue (DAT) has received lots of attention as biological scaffolds recently. DAT that extracted from the extracellular matrix (ECM) of adipose tissues holds great promise as a xenogeneic biomaterial for tissue engineering and regenerative medicine. In our study, 2-D DATsol film was fabricated to enhance cell adhesion, proliferation, and differentiation of ASCs in vitro. DAT was also used to modify alginate for improvement of cell adhesion. Alginate microspheres modified with DAT were prepared by Nisco. These microspheres could provide a highly supportive 3-D environment for ASCs. In our works, ASCs were immobilized in alginate microspheres modified with DAT to promoted cell adhesion and adipogenic differentiation. Accordingly, we hypothesize that tissue regeneration in vivo could be promoted with the aid of modified microspheres in future.

Keywords: adipose stem cells, decellularize adipose tissue, Alginate, microcarries

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Authors: Sarwar Beg, Gajanand Sharma, O. P. Katare, Bhupinder Singh

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The current studies entail development of self-nano emulsifying drug delivery systems (SNEDDS) of rosuvastatin, employing rational QbD-based approach for enhancing its oral bioavailability. SNEDDS were prepared using the blend of lipidic and emulsifying excipients, i.e., Peceol, Tween 80, and Transcutol HP. The prepared formulations evaluated for in vitro drug release, ex vivo permeation, in situ perfusion studies and in vivo pharmacokinetic studies in rats, which demonstrated 3-4 fold improvement in biopharmaceutical performance of the developed formulations. Cytotoxicity studies using MTT assay and histopathological studies in intestinal cells revealed the lack of cytotoxicity and thereby safety and efficacy of the developed formulations.

Keywords: SNEDDS, bioavailability, solubility, Quality by Design (QbD)

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Authors: Maria E. V. Amarante, Carolina L. Cerdeira, Julia V. Cortes, Fiorita G. L. Mundim

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Basal cell carcinoma (BCC) is the most prevalent type of skin cancer in humans. It arises from the basal cells of the epidermis and cutaneous appendages. The role of sunlight exposure as a risk factor for BCC is very well defined due to its power to influence genetic mutations, in addition to having a suppressor effect on the skin immune system. Despite showing low metastasis and mortality rates, the tumor is locally infiltrative, aggressive, and destructive. Considering the high prevalence rate of this carcinoma and the importance of early detection, a retrospective study was carried out in order to correlate the clinical data available on BBC, characterize it epidemiologically, and thus enable effective prevention measures for the population. Data on the period from January 2015 to December 2019 were collected from the medical records of patients registered at one pathology service located in the southeast region of Brazil, known as SVO, which delivers skin biopsy results. The study was aimed at correlating the variables, sex, age, and subtypes found. Data analysis was performed using the chi-square test at a nominal significance level of 5% in order to verify the independence between the variables of interest. Fisher's exact test was applied in cases where the absolute frequency in the cells of the contingency table was less than or equal to five. The statistical analysis was performed using the R® software. Ninety-three basal cell carcinoma were analyzed, and its frequency in the 31-to 45-year-old age group was 5.8 times higher in men than in women, whereas, from 46 to 59 years, the frequency was found 2.4 times higher in women than in men. Between the ages of 46 to 59 years, it should be noted that the sclerodermiform subtype appears more than the solid one, with a difference of 7.26 percentage points. Reversely, the solid form appears more frequently in individuals aged 60 years or more, with a difference of 8.57 percentage points. Among women, the frequency of the solid subtype was 9.93 percentage points higher than the sclerodermiform frequency. In males, the same percentage difference is observed, but sclerodermiform is the most prevalent subtype. It is concluded in this study that, in general, there is a predominance of basal cell carcinoma in females and in individuals aged 60 years and over, which demonstrates the tendency of this tumor. However, when rarely found in younger individuals, the male gender prevailed. The most prevalent subtype was the solid one. It is worth mentioning that the sclerodermiform subtype, which is more aggressive, was seen more frequently in males and in the 46-to 59-year-old range.

Keywords: basal cell carcinoma, epidemiology, sclerodermiform basal cell carcinoma, skin cancer, solar radiation, solid basal cell carcinoma

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Authors: Sarah Crawford, Gerry Lesley

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This research is a pre-clinical assessment of anti-cancer effects of novel non-steroidal diethyl stilbestrol-like estrogen analogs in estrogen-receptor positive/ progesterone-receptor positive human breast cancer microtumors of MCF 7 cell line. Tamoxifen analog formulation (Tam A1) was used as a single agent or in combination with therapeutic concentrations of 4-hydroxytamoxifen, currently used as a long-term treatment for the prevention of breast cancer recurrence in women with estrogen receptor positive/ progesterone receptor positive malignancies. At concentrations ranging from 30-50 microM, Tam A1 induced microtumor disaggregation and cell death. Incremental cytotoxic effects correlated with increasing concentrations of Tam A1. Live tumor microscopy showed that microtumos displayed diffuse borders and substrate-attached cells were rounded-up and poorly adherent. A complete cytotoxic effect was observed using 40-50 microM Tam A1 with time course kinetics similar to 4-hydroxytamoxifen. Combined treatment with TamA1 (30-50 microM) and 4-hydroxytamoxifen (10-15 microM) induced a highly cytotoxic, synergistic combined treatment response that was more rapid and complete than using 4-hydroxytamoxifen as a single agent therapeutic. Microtumors completely dispersed or formed necrotic foci indicating a highly cytotoxic combined treatment response. Moreover, breast cancer microtumors treated with both 4-hydroxytamoxifen and Tam A1 displayed lower levels of long-term post-treatment regrowth, a critical parameter of primary drug resistance, than observed for 4-hydroxytamoxifen when used as a single agent therapeutic. Tumor regrowth at 6 weeks post-treatment with either single agent 4-hydroxy tamoxifen, Tam A1 or a combined treatment was assessed for the development of drug resistance. Breast cancer cells treated with both 4-hydroxytamoxifen and Tam A1 displayed significantly lower levels of post-treatment regrowth, indicative of decreased drug resistance, than observed for either single treatment modality. The preclinical data suggest that combined treatment involving the use of tamoxifen analogs may be a novel clinical approach for long-term maintenance therapy in patients with estrogen-receptor positive/progesterone-receptor positive breast cancer receiving hormonal therapy to prevent disease recurrence. Detailed data on time-course, IC50 and tumor regrowth assays post- treatment as well as a proposed mechanism of action to account for observed synergistic drug effects will be presented.

Keywords: 4-hydroxytamoxifen, tamoxifen analog, drug-resistance, microtumors

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Authors: Ananya Pal, Neelakshi Sarkar, Debraj Saha, Dipanwita Das, Subhashish Kamal Guha, Bibhuti Saha, Runu Chakravarty

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Presently, tenofovir disoproxil fumurate (TDF) is the most effective anti-viral agent for the treatment of hepatitis B virus (HBV) in individuals co-infected with HIV and HBV as TDF has activity to suppress both wild-type and lamivudine (3TC)-resistant HBV. However, suboptimal response to TDF was reported in HIV-HBV co-infected individuals with prior 3TC therapy from different countries recently. The incidence of 3TC-resistant HBV strains is quite high in HIV-HBV co-infected patients experiencing long-term anti-retroviral therapy (ART) in eastern India. In spite of this risk, most of the patients with long-term 3TC treatment are continued with the same anti-viral agent in this country. Only a few have received TDF in addition to 3TC in the ART regimen since TDF has been available in India for the treatment of HIV-infected patients in 2012. In this preliminary study, we investigated the virologic and biochemical parameters among HIV-HBV co-infected patients who are non-responders to 3TC treatment during the continuation of 3TC or TDF add-on to 3TC in their ART regimen. Fifteen HIV-HBV co-infected patients who experienced long-term 3TC (mean duration months 36.87 ± 24.08 months) were identified with high HBV viremia ( > 20,000 IU/ml) or harbouring 3TC-resistant HBV. These patients receiving ART from School of Tropical Medicine Kolkata, the main ART centre in eastern India were followed-up semi-annually for next three visits. Different virologic parameters including quantification of plasma HBV load by real-time PCR, detection of hepatitis B e antigen (HBeAg) by commercial ELISA and anti-viral resistant mutations by sequencing were studied. During three follow-up among study subjects, 86%, 47%, and 43% had 3TC-mono-therapy (mean treatment-duration 41.54±18.84, 49.67±11.67, 54.17±12.37 months respectively) whereas 14%, 53%, and 57% experienced TDF in addition to 3TC (mean treatment duration 4.5±2.12, 16.56±11.06, and 23±4.07 months respectively). Mean CD4 cell-count in patients receiving 3TC was tended to be lower during third follow-up as compared to the first and the second [520.67±380.30 (1st), 454.8±196.90 (2nd), and 397.5±189.24 (3rd) cells/mm3) and similar trend was seen in patients experiencing TDF in addition to 3TC [334.5±330.218 (1st), 476.5±194.25 (2nd), and 461.17±269.89 (3rd) cells/mm3]. Serum HBV load was increased during successive follow-up of patients with 3TC-mono-therapy. Initiation of TDF lowered serum HBV-load among 3TC-non-responders at the time of second visit ( < 2,000 IU/ml), interestingly during third follow-up, mean HBV viremia increased >1 log IU/ml (mean 3.56±2.84 log IU/ml). Persistence of 3TC-resistant double and triple mutations was also observed in both the treatment regimens. Mean serum alanine aminotransferase remained elevated in these patients during this follow-up study. Persistence of high HBV viraemia and 3TC-resistant mutation in HBV during the continuation of 3TC might lead to major public health threat in India. The inclusion of TDF in the ART regimen of 3TC non-responder HIV-HBV co-infected patients showed adverse treatment response in terms of virologic and biochemical parameters. Therefore, serious attention is necessary for proper management of long-term 3TC experienced HIV-HBV co-infected patients with high HBV viraemia or 3TC-resistant HBV mutants in India.

Keywords: HBV, HIV, TDF, 3TC-resistant

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Authors: Imran Ali

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General chemotherapy for cancer treatment has many side and toxic effects. A new approach of targeting nano anti-cancer drug is under development stage and only few drugs are available in the market today. The unique features of these drugs are targeted action on cancer cells only without any side effect. Sometimes, these are called magic drugs. The important molecules used for nano anti-cancer drugs are cisplatin, carboplatin, bleomycin, 5-fluorouracil, doxorubicin, dactinomycin, 6-mercaptopurine, paclitaxel, topotecan, vinblastin and etoposide etc. The most commonly used materials for preparing nano particles carriers are dendrimers, polymeric, liposomal, micelles inorganic, organic etc. The proposed lecture will comprise the-of-art of nano drugs in cancer chemo-therapy including preparation, types of drugs, mechanism, future perspectives etc.

Keywords: cancer, nano-anti-cancer drugs, chemo-therapy, mechanism of action, future perspectives

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Authors: Yener Tekeli, Hayri Baba

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The natural active compounds found in medicinal plants are belong to various chemical structures including polyphenolic compounds, flavonoids, essential oils, and vitamins and some of these compounds have anticancer, antioxidant, and antimicrobial activity. However, these compounds have been little known about mechanisms to confer antibacterial drug resistance. In this study; some macrofungi extracts (Pholiota lucifera, Gnaoderma applanatum and Pleurotus ostreatus) were investigated for their abilities to enhance bacterial permeability by flow cytometry. This experiments exhibited enhancement of these extracts to disrupt the cytoplasmic membrane of living bacterial (Listeria innocua and Escherichia coli) cells. These experiments were designed to detect uptake of PI&SYT by enhancing with a ranged concentration of herb extracts.

Keywords: antimicrobial activity, flow cytometry, macrofungi, multidrug resistant

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Authors: Kunal Das, Priya Sengupta, Abhishek K. Singh

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Pin assignment, scheduling, routing and error detection for arbitrary biochemical protocols in Digital Microfluidic Biochip have been reported in this paper. The research work is concentrating on pin assignment for 2 or 3 droplets routing in the arbitrary biochemical protocol, scheduling and routing in m × n biochip. The volumetric error arises due to droplet split in the biochip. The volumetric error detection is also addressed using biochip AND logic gate which is known as microfluidic AND or mAND gate. The algorithm for pin assignment for m × n biochip required m+n-1 numbers of pins. The basic principle of this algorithm is that no same pin will be allowed to be placed in the same column, same row and diagonal and adjacent cells. The same pin should be placed a distance apart such that interference becomes less. A case study also reported in this paper.

Keywords: digital microfludic biochip, cross-contamination, pin assignment, microfluidic AND gate

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Authors: Chien-Liang Chao, Yun-Sheng Lin

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Secondary metabolites are applied in the human life of the Chinese herbal medicine. Many drugs are originally extracted from natural products with combination of pharmaceutical and chemical studies. Crude extract of the leaves from Zamia furfureace L. has been shown to exhibit anticancer activities. The first chemical investigation of this plant was carried out by our group. In this study, four known compounds were isolated from Zamia furfureace L. with three lignins (Sesamin (1), Wodeshiol (2) and Paulownin (3)), and one dipeptide (Aurantiamide acetate (4)). The structures of these compounds were analyzed through the 1D-NMR(1H-NMR,13C-NMR)、2D-NMR(COSY、HMQC、HMBC、NOESY) spectroscopic analysis, and by comparison of variety of physical data (IR, mass spectrometry, ultraviolet, optical rotation). Among them, Aurantiamide acetate (4) exhibited weak cytotoxic activity against human gastric cancer cells.

Keywords: Zamia furfureace L., AGS, sesamin, Aurantiamide acetate, secondary metabolites

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Authors: Darshna Sharma, Suban K. Sahoo

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The detection of intracellular fluoride in human cancer cell HeLa was achieved by chemosensors derived from vitamin B6 cofactors using fluorescence imaging technique. These sensors were first synthesized by condensation of pyridoxal/pyridoxal phosphate with 2-amino(thio)phenol. The anion recognition ability was explored by experimental (UV-VIS, fluorescence and 1H NMR) and theoretical DFT [(B3LYP/6-31G(d,p)] methods in DMSO and mixed DMSO-H2O system. All the developed sensors showed both naked-eye detectable color change and remarkable fluorescence enhancement in the presence of F- and AcO-. The anion recognition was occurred through the formation of hydrogen bonded complexes between these anions and sensor, followed by the partial deprotonation of sensor. The detection limit of these sensors were down to micro(nano) molar level of F- and AcO-.

Keywords: chemosensors, fluoride, acetate, turn-on, live cells imaging, DFT

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Authors: David Oluigbo, Erik Hemberg, Nathan Shwatal, Wenqi Ding, Yin Yuan, Susanna Mierau

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Introduction: Calcium imaging is an established technique in neuroscience research for detecting activity in neural networks. Bursts of action potentials in neurons lead to transient increases in intracellular calcium visualized with fluorescent indicators. Manual identification of cell bodies and their contours by experts typically takes 10-20 minutes per calcium imaging recording. Our aim, therefore, was to design an automated pipeline to facilitate and optimize calcium imaging data analysis. Our pipeline aims to accelerate cell body and contour identification and production of graphical representations reflecting changes in neuronal calcium-based fluorescence. Methods: We created a Python-based pipeline that uses OpenCV (a computer vision Python package) to accurately (1) detect neuron contours, (2) extract the mean fluorescence within the contour, and (3) identify transient changes in the fluorescence due to neuronal activity. The pipeline consisted of 3 Python scripts that could both be easily accessed through a Python Jupyter notebook. In total, we tested this pipeline on ten separate calcium imaging datasets from murine dissociate cortical cultures. We next compared our automated pipeline outputs with the outputs of manually labeled data for neuronal cell location and corresponding fluorescent times series generated by an expert neuroscientist. Results: Our results show that our automated pipeline efficiently pinpoints neuronal cell body location and neuronal contours and provides a graphical representation of neural network metrics accurately reflecting changes in neuronal calcium-based fluorescence. The pipeline detected the shape, area, and location of most neuronal cell body contours by using binary thresholding and grayscale image conversion to allow computer vision to better distinguish between cells and non-cells. Its results were also comparable to manually analyzed results but with significantly reduced result acquisition times of 2-5 minutes per recording versus 10-20 minutes per recording. Based on these findings, our next step is to precisely measure the specificity and sensitivity of the automated pipeline’s cell body and contour detection to extract more robust neural network metrics and dynamics. Conclusion: Our Python-based pipeline performed automated computer vision-based analysis of calcium image recordings from neuronal cell bodies in neuronal cell cultures. Our new goal is to improve cell body and contour detection to produce more robust, accurate neural network metrics and dynamic graphs.

Keywords: calcium imaging, computer vision, neural activity, neural networks

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Authors: Gustavo Axel Elizalde-Velázquez

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Metformin is the first line of oral therapy to treat type II diabetes and is also employed as a treatment for other indications, such as polycystic ovary syndrome, cancer, and COVID-19. Recent data suggest it is the aspirin of the 21st century due to its antioxidant and anti-aging effects. However, increasingly current articles indicate its long-term consumption generates mitochondrial impairment. Up to date, it is known metformin increases the biogenesis of Alzheimer's amyloid peptides via up-regulating BACE1 transcription, but further information related to brain damage after its consumption is missing. Bearing in mind the above, this work aimed to establish whether or not chronic exposure to metformin may alter swimming behavior and induce neurotoxicity in Danio rerio adults. For this purpose, 250 Danio rerio grown-ups were assigned to six tanks of 50 L of capacity. Four of the six systems contained 50 fish, while the remaining two had 25 fish (≈1 male:1 female ratio). Every system with 50 fish was allocated one of the three metformin treatment concentrations (1, 20, and 40 μg/L), with one system as the control treatment. Systems with 25 fish, on the other hand, were used as positive controls for acetylcholinesterase (10 μg/L of Atrazine) and oxidative stress (3 μg/L of Atrazine). After four months of exposure, a mean of 32 fish (S.D. ± 2) per group of MET treatment survived, which were used for the evaluation of behavior with the Novel Tank test. Moreover, after the behavioral assessment, we aimed to collect the blood and brains of all fish from all treatment groups. For blood collection, fish were anesthetized with an MS-222 solution (150 mg/L), while for brain gathering, fish were euthanized using the hypothermic shock method (2–4 °C). Blood was employed to determine CASP3 activity and the percentage of apoptotic cells with the TUNEL assay, and brains were used to evaluate acetylcholinesterase activity, oxidative damage, and gene expression. After chronic exposure, MET-exposed fish exhibited less swimming activity when compared to control fish. Moreover, compared with the control group, MET significantly inhibited the activity of AChE and induced oxidative damage in the brain of fish. Concerning gene expression, MET significantly upregulated the expression of Nrf1, Nrf2, BAX, p53, BACE1, APP, PSEN1, and downregulated CASP3 and CASP9. Although MET did not overexpress the CASP3 gene, we saw a meaningful rise in the activity of this enzyme in the blood of fish exposed to MET compared to the control group, which we then confirmed by a high number of apoptotic cells in the TUNEL assay. To the best of our understanding, this is the first study that delivers evidence of oxidative impairment, apoptosis, AChE alteration, and overexpression of B- amyloid-related genes in the brain of fish exposed to metformin.

Keywords: AChE inhibition, CASP3 activity, NovelTank test, oxidative damage, TUNEL assay

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Authors: Miguel Contreras, David Long, Will Bachman

Abstract:

Background: Microscopy plays a central role in cell and developmental biology. In particular, fluorescence microscopy can be used to visualize specific cellular components and subsequently quantify their morphology through development of virtual-cell models for study of effects of mechanical forces on cells. However, there are challenges with these imaging experiments, which can make it difficult to quantify cell morphology: inconsistent results, time-consuming and potentially costly protocols, and limitation on number of labels due to spectral overlap. To address these challenges, the objective of this project is to develop an automatic computational machine learning pipeline to predict cellular components morphology for virtual-cell generation based on fluorescence cell membrane confocal z-stacks. Methods: Registered confocal z-stacks of nuclei and cell membrane of endothelial cells, consisting of 20 images each, were obtained from fluorescence confocal microscopy and normalized through software pipeline for each image to have a mean pixel intensity value of 0.5. An open source machine learning algorithm, originally developed to predict fluorescence labels on unlabeled transmitted light microscopy cell images, was trained using this set of normalized z-stacks on a single CPU machine. Through transfer learning, the algorithm used knowledge acquired from its previous training sessions to learn the new task. Once trained, the algorithm was used to predict morphology of nuclei using normalized cell membrane fluorescence images as input. Predictions were compared to the ground truth fluorescence nuclei images. Results: After one week of training, using one cell membrane z-stack (20 images) and corresponding nuclei label, results showed qualitatively good predictions on training set. The algorithm was able to accurately predict nuclei locations as well as shape when fed only fluorescence membrane images. Similar training sessions with improved membrane image quality, including clear lining and shape of the membrane, clearly showing the boundaries of each cell, proportionally improved nuclei predictions, reducing errors relative to ground truth. Discussion: These results show the potential of pre-trained machine learning algorithms to predict cell morphology using relatively small amounts of data and training time, eliminating the need of using multiple labels in immunofluorescence experiments. With further training, the algorithm is expected to predict different labels (e.g., focal-adhesion sites, cytoskeleton), which can be added to the automatic machine learning pipeline for direct input into Principal Component Analysis (PCA) for generation of virtual-cell mechanical models.

Keywords: cell morphology prediction, computational machine learning, fluorescence microscopy, virtual-cell models

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Authors: Mayuri Gandhi, Nayan Agrawal, Harshita Bhatia

Abstract:

In the present work, cerium fluoride (CeF3) was selected as the host material because of its high density, fast response and high radiation resistance, efficient absorption and energy transfer by host (to activator). For the synthesis of CeF3 nanoparticles doped with Sm3+ ion, co-precipitation route was employed. Thus for optimum results, concentration dependent studies of the fluorescence of Sm3+ was carried out. The photoluminescence gave emissions in both visible as well as the NIR region and therefore it can have its application in solar cells, where it can absorb a large spectrum of energy. CeF3:Sm3+ nanoparticles were carefully incorporated in a suitable polymer matrix in order to demonstrate a variety of applications to improve the performance of the polymer materials and use it to develop high grade optoelectronic devices such as LEDs, security labelling, lasers, displays, biological imaging, etc.

Keywords: bioimaging, cerium fluoride, NIR emission, samarium

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Authors: Asowata Osamede, Trudy Sutherland

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The purpose of this paper is to assess the available power from a Renewable Energy Source (off-grid photovoltaic (PV) panel) in the Southern Hemisphere using anisotropic model. Direct solar radiation is the driving force in photovoltaics. In a basic PV panels in the Southern Hemisphere, Power conversion is eminent, and this is achieved by the PV cells converting solar energy into electrical energy. In this research, the results was determined for a 6 month period from September 2022 through February 2023. Preliminary results, which include Normal Probability plot, data analysis - R2 value, effective conversion-time per week and work-time per day, indicate a favorably comparison between the empirical results and the simulation results.

Keywords: power-conversion, mathematical model, PV panels, DC-DC converters, direct solar radiation

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Authors: Karim Matter, Heba Fayed, Ahmed Abd-Elaziz, Moustafa Hussein

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Due to the exponential growth of mobile devices and wireless services, a huge demand for radiofrequency has increased. The presence of several frequencies causes interference between cells, which must be minimized to get the lower Bit Error Rate (BER). For this reason, it is of great interest to use visible light communication (VLC). This paper suggests a VLC system that decreases the BER by applying a new LED distribution with a hexagonal shape using a Frequency Reuse (FR) concept to mitigate the interference between the reused frequencies inside the hexagonal shape. The BER is measured in two scenarios, Line of Sight (LoS) and Non-Line of Sight (Non-LoS), for each technique that we used. The recommended values of BER in the proposed model for Soft Frequency Reuse (SFR) in the case of Los at 4, 8, and 10 dB signal to noise ratio (SNR), are 3.6×10⁻⁶, 6.03×10⁻¹³, and 2.66×10⁻¹⁸, respectively.

Keywords: visible light communication (VLC), field of view (FoV), hexagonal array, frequency reuse

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Authors: Vanessa Funk, Steven Blum, Stephanie Cole, Jorge Maciel, Matthew Lux

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Paper-based synthetic gene circuits offer a new paradigm for programmable, fieldable biodetection. We demonstrate that by freeze-drying gene circuits with in vitro expression machinery, we can use complimentary RNA sequences to trigger colorimetric changes upon rehydration. We have successfully utilized both green fluorescent protein and luciferase-based reporters for easy visualization purposes in solution. Through several efforts, we are aiming to use this new platform technology to address a variety of needs in portable detection by demonstrating several more expression and reporter systems for detection functions on paper. In addition to RNA-based biodetection, we are exploring the use of various mechanisms that cells use to respond to environmental conditions to move towards all-hazards detection. Examples include explosives, heavy metals for water quality, and toxic chemicals.

Keywords: cell-free lysates, detection, gene circuits, in vitro

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Authors: Abderrazak Bannari, Ghadeer Kadhem

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The objectives of this paper are the validation and the evaluation of MBES-CARIS BASE surface data performance for bathymetric mapping of shallow water in the Kingdom of Bahrain. The latter is an archipelago with a total land area of about 765.30 km², approximately 126 km of coastline and 8,000 km² of marine area, located in the Arabian Gulf, east of Saudi Arabia and west of Qatar (26° 00’ N, 50° 33’ E). To achieve our objectives, bathymetric attributed grid files (X, Y, and depth) generated from the coverage of ship-track MBSE data with 300 x 300 m cells, processed with CARIS-HIPS, were downloaded from the General Bathymetric Chart of the Oceans (GEBCO). Then, brought into ArcGIS and converted into a raster format following five steps: Exportation of GEBCO BASE surface data to the ASCII file; conversion of ASCII file to a points shape file; extraction of the area points covering the water boundary of the Kingdom of Bahrain and multiplying the depth values by -1 to get the negative values. Then, the simple Kriging method was used in ArcMap environment to generate a new raster bathymetric grid surface of 30×30 m cells, which was the basis of the subsequent analysis. Finally, for validation purposes, 2200 bathymetric points were extracted from a medium scale nautical map (1:100 000) considering different depths over the Bahrain national water boundary. The nautical map was scanned, georeferenced and overlaid on the MBES-CARIS generated raster bathymetric grid surface (step 5 above), and then homologous depth points were selected. Statistical analysis, expressed as a linear error at the 95% confidence level, showed a strong correlation coefficient (R² = 0.96) and a low RMSE (± 0.57 m) between the nautical map and derived MBSE-CARIS depths if we consider only the shallow areas with depths of less than 10 m (about 800 validation points). When we consider only deeper areas (> 10 m) the correlation coefficient is equal to 0.73 and the RMSE is equal to ± 2.43 m while if we consider the totality of 2200 validation points including all depths, the correlation coefficient is still significant (R² = 0.81) with satisfactory RMSE (± 1.57 m). Certainly, this significant variation can be caused by the MBSE that did not completely cover the bottom in several of the deeper pockmarks because of the rapid change in depth. In addition, steep slopes and the rough seafloor probably affect the acquired MBSE raw data. In addition, the interpolation of missed area values between MBSE acquisition swaths-lines (ship-tracked sounding data) may not reflect the true depths of these missed areas. However, globally the results of the MBES-CARIS data are very appropriate for bathymetric mapping of shallow water areas.

Keywords: bathymetry mapping, multibeam echosounder systems, CARIS-HIPS, shallow water

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Authors: Mohamed El Hachimi, Abdelhakim Ballouk, Ilyas Khelafa, Abdelaziz Mouhou

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Introduction: Type 1 diabetes occurs when beta cells are destroyed by the body's own immune system. Treatment of type 1 diabetes mellitus could be greatly improved by applying a closed-loop control strategy to insulin delivery, also known as an Artificial Pancreas (AP). Method: In this paper, we present a new formulation of the cost function for a Model Predictive Control (MPC) utilizing a technic which accelerates the speed of control of the AP and tackles the nonlinearity of the control problem via asymmetric objective functions. Finding: The finding of this work consists in a new Model Predictive Control algorithm that leads to good performances like decreasing the time of hyperglycaemia and avoiding hypoglycaemia. Conclusion: These performances are validated under in silico trials.

Keywords: artificial pancreas, control algorithm, biomedical control, MPC, objective function, nonlinearity

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Authors: Beata Jackowska-Zduniak

Abstract:

We formulate and analyze a mathematical model describing dynamics of cancer growth under the influence of radiation therapy. The effect of this type of therapy is considered as an additional equation of discussed model. Numerical simulations show that delay, which is added to ordinary differential equations and represent time needed for transformation from one type of cells to the other one, affects the behavior of the system. The validation and verification of proposed model is based on medical data. Analytical results are illustrated by numerical examples of the model dynamics. The model is able to reconstruct dynamics of treatment of cancer and may be used to determine the most effective treatment regimen based on the study of the behavior of individual treatment protocols.

Keywords: mathematical modeling, numerical simulation, ordinary differential equations, radiation therapy

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Authors: Asmat Nawaz, Ali Koray Erdinc, Burak Gultekin, Muhammad Tayyib, Ceylan Zafer, Kaiying Wang, M. Nadeem Akram

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In this study, a sequential deposition process is used for the fabrication of PEDOT: PSS based inverted planar perovskite solar cell. A small amount of additive deionized water (DI-H₂O) was added into PbI₂ + Dimethyl formamide (DMF) precursor solution in order to increase the solubility of PbI₂ in DMF, and finally to manipulate the surface morphology of the perovskite films. A morphology transition from needle like structure to hexagonal plates, and then needle-like again has been observed as the DI-H2O was added continuously (0.0 wt% to 3.0wt%). The latter one leads to full surface coverage of the perovskite, which is essential for high performance solar cell.

Keywords: charge carrier diffusion lengths, Methylamonium lead iodide, precursor composition, perovskite solar cell, sequential deposition

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Authors: Jasmeet Kaur, Anup Singh, Shashikant Iyengar, Arun Kumar, Ira Sahay

Abstract:

Latent autoimmune diabetes in adults (LADA) is an irreversible autoimmune disease that affects insulin production. LADA is characterized by the production of Glutamic acid decarboxylase (GAD) antibodies, which is similar to type 1 diabetes. Individuals with LADA may eventually develop overt diabetes and require insulin. In this condition, the pancreas produces little or no insulin, which is a hormone used by the body to allow glucose to enter cells and produce energy. While type 1 diabetes was traditionally associated with children and teenagers, its prevalence has increased in adults as well. LADA is frequently misdiagnosed as type 2 diabetes, especially in adulthood when type 2 diabetes is more common. LADA develops in adulthood, usually after age 30. Managing LADA involves metabolic control with exogenous insulin and prolonging the life of surviving beta cells, thereby slowing the disease's progression. This case study examines the impact of approximately 3 months of low-carbohydrate dietary intervention in a 42-year-old woman with LADA who was initially misdiagnosed as having type 2 diabetes. Her c-peptide was 0.13 and her HbA1c was 9.3% when this trial began. Low-carbohydrate interventions have been shown to improve blood sugar levels, including fasting, post-meal, and random blood sugar levels, as well as haemoglobin levels, blood pressure, energy levels, sleep quality, and satiety levels. The use of low-carbohydrate dietary intervention significantly reduces both hypo- and hyperglycaemia events. During the 3 months of the study, there were 2 to 3 hyperglycaemic events owing to physical stress and a single hypoglycaemic event. Low-carbohydrate dietary therapies lessen insulin dose inaccuracy, which explains why there were fewer hyperglycaemic and hypoglycaemic events. In three months, the glycated haemoglobin (HbA1c) level was reduced from 9.3% to 6.3%. These improvements occur without the need for caloric restriction or physical activity. Stress management was crucial aspect of the treatment plan as stress-induced neuroendocrine hormones can cause immunological dysregulation. Additionally, supplements that support immune system and reduce inflammation were used as part of the treatment during the trial. Long-term studies are needed to track disease development and corroborate the claim that such dietary treatments can prolong the honeymoon phase in LADA. Various factors can contribute to additional autoimmune attacks, so measuring c-peptide is crucial on a regular basis to determine whether insulin levels need to be adjusted.

Keywords: autoimmune, diabetes, LADA, low_carb, nutrition

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