Analysis of Oyster Plant (Tradescantia Spathacea) Extracts via Maceration, Soxhlet Extraction and Thin Layer Chromatograpy

Daniel Russo, Cristina Balistreri, Kelly O’Reilly, Luis Cendan, Carlos Vazquez, Pilar Maul and Maria Pina

School of Science, Technology and Engineering Management, St. Thomas University, Miami Gardens, FL 33054.

The Oyster Plant (Tradescantia Spathacea) is a fleshy or succulent perennial garden herb. This plant is an ornamental plant and found in many tropical countries. Medicinally, the plant is used for colds, sore throat, whooping cough, nasal bleeding, and also as an anti-inflammatory. The plant was grown in the organic garden at St. Thomas University and collected there. All of the plant parts were separated and cleaned up first, and divided into its different parts: leaves, stems, roots, and flowers. The parts were dried two days at 40°C and grinded for wet and dry extraction. The methods included maceration and Soxhlet extraction. Maceration is a type of extraction where the plant materials were placed in different mixtures of solvents, stirred so plant material can be extracted, and left to sit for number of days at room temperature. In the Soxhlet extraction the solvent is heated to reflux, the vapor travels up a distillation arm, and floods into the chamber housing the thimble of solid. The condenser ensures that any solvent vapor cools, and drips back down into the chamber housing the solid material. Soxhlet extraction is done taking the dry material that can be repeatedly washed by a solvent. All the extracts were rot vapored and analyzed using Thin Layer Chromatography with polar and nonpolar solvents. The spots were developed and visualized with iodine and UV light. It has been found that the roots and leaves contain polar and non polar organic compounds The present work reports the best solvents for extraction, and the better separation was found to be a mixture of 75% ethanol and 25% hexane and 50% ethanol and 50% hexane. Preliminary analysis of extracts using column chromatography and test for inhibition of cancer cell growth were initiated

Culturing Human Neural Stem Cells and Quantifying Lentiviruses to Study Autism.

Milagros Mulero¹, Leana Ramos¹, Vadym Trokhymchuk¹, Deliabell Hernandez¹, Alexis Tapanes-Castillo¹, Derek Dyxkhoorn²

¹School of Science, Technology, and Engineering Management, St. Thomas University, Miami Gardens, FL 33054; ²Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, FL 33136

 Autism spectrum disorder (ASD) is partly caused by problems in neuronal communication. Differences in the RNA expression levels of the synaptic protein Discs large homolog-associated protein 1 (DLGAP1) have recently been associated with ASD. The long term goal of this project is to analyze how human neurons react to reduced DLGAP1 antisense RNA levels. To accomplish this, we cultured neural progenitor cells (NPCs) from autistic and control patients. These neural progenitor cells, which were derived from induced pluripotent stem cells that originated from patient skin biopsies, were differentiated towards a glutamatergic fate. In separate experiments, done at the University of Miami, six lentiviruses designed to degrade DLGAP1 antisense RNA transcripts were produced. By generating a standard curve, we quantified the number of virus particles present in each sample. In the future, autistic and control neural progenitor cells will be infected with lentiviruses to analyze how reducing DLGAP1 antisense RNA levels affects autistic cells.

Flavonoid compounds are normally present on nature being a common constituent of medicinal plants. These natural products have several health benefits, including activity against tropical parasites, such as Trypanosoma cruzi and Leishmania spp. Five flavonoid active against Trypanosoma cruzi and five active against Leishmania species were submitted to molecular docking with the enzymes cruzain and trypanothione reductase from Trypanosoma cruzi and N-myristoyltransferase, dihydroorotate dehydrogenase and trypanothiona reductase from Leishmania spp. Considering the binding energies and interactions between both, compounds and enzymes, the flavonoids chrysin dimethylether  and tamarixetin against T. cruzi, and luteolin and ladanein against Leishmania spp presented the best results. This study hopes to contribute to existing research on these natural products against these tropical parasites.

Pharmacological receptors are important because they produce an interaction with drugs leading to a pharmacological response. The β₁ and β₂ adrenergic receptors are found in cardiac muscle and bronchial tissue, as well as cardiomyocytes. The family Menispermaceae comprises over 70 genera with a wide global distribution, many having different pharmacological activities. Several secondary metabolites, particularly alkaloids, have been isolated from this family. The aim of this study was to evaluate a number of alkaloids and their affinity to β₁ and β₂ adrenergic receptors. Therefore, applied-based virtual screening of ligands associated with screening based on the virtual structure of a small data set of 786 secondary metabolites of the Menispermaceae family from an in-house database was undertaken. The multitarget selectivity of the alkaloid dauricine (482) was observed for inhibiting β₁ adrenergic receptors and activating β₂ adrenergic receptors. These compounds can be used as starting points for studies of proposed structures for cardioprotective and/or antiasthmatic activity.

Biologically active molecules present in the venom of several species of scorpion, have shown potential against various diseases, including cancer. It has been reported that several toxins can block ion channels present in the membrane of several cancer cell lines, through action potential, altering their cell function, cell cycle arresting, and inducing apoptosis death pathway. The use of the sequence of Cll-6 gene present in the genome of the Mexican scorpion Centruroides limpidus limpidus encoding a beta-toxin-locker of Na + channels by action potentials is proposed. This molecule has not been evaluated to determine if it has potential as an anti-cancer agent. In this paper the in silico design of recombinant molecule pDream2.1/MCS/CII-6, including the CII-6 gene into the polylinker site and identification by molecular biology techniques. Our results confirm a fragment of 316 base pairs, by digestion with BamH1 and Kpn1 enzymes, PCR, and the sequenced of amplicon, the alignment with CII-6 sequence reported in the GenBank database, showed 99.6% similarity and identity. The recombinant plasmid, could be used to assess potential as anti-having cancer agent on several cancer cell lines.

Canine parvovirus type 2 (CPV-2) is the main etiological agent of viral enteritis in dogs. Actually in literature, CPV-2 has been reported with clinical signs that vary from the classical disease. In this study, we evaluated the clinical signs presented in 50 dogs infected naturally with CPV-2. All the infected dogs were analyzed by PCR and sequenced. Our data indicate that the CPV-2c is the most frequently genovariant in Mexico the 50 dogs belong to the CPV-2c, through the amino acid change 426 Asn-Glu and concerning clinical signs, the presence of either vomiting or enlarged lymph node was observed in all virally infected patients.

Chagas disease, or American trypanosomiasis, is caused by Trypanosoma cruzi, a hemoflagellate Trypanosomatidae family parasite, widely distributed in the American tropics and subtropics. Cruzipain (Cz) is a crucial parasitic proteases belong to the papain-like CA C1 family and have close structural mammalian homologues and considered to be a good vaccine candidate for Chagas disease. Cz full gene sequence was obtained from GenBank Acc. No. AY099317.1 and redesigned in silico, sequences were sent to be synthesized and cloned into Sc expression plasmids (pYES-2, Invitrogen) in Genscript Company (USA). pYES-CZ recombinant plasmid was transformed into DH5α E.coli strain for its selection and production. Plasmidic DNA was extracted and transfected to INVSc1 Saccharomyces cerevisiae strain, then cultivated and selected through ura3-52 deficient medium. Finally, transfected Sc were evaluated as a potential vaccine against Chagas disease in 8 weeks female BALB/c mice. Results showed that modified cruzipain gene construct was successfully stable during its transfection in bacteria then in Sc, moreover amplification results showed a unified linear DNA with the desired size. Data showed that transfected recombinant Sc elevates lymphocyte responses and gives remarkable results. Lastly Sc and Cz are promising vaccine candidates against T. cruzi experimental infection, further investigation would be required for establishing a strong therapy for the population at risk of the infection.

Immunohistochemistry revealed by diaminobenzidine (IHC-DAB) or fluorescence (IHC-F) are two of the most common techniques used in histopathology. In order to evaluate the similarity between two methodologies that share the same antigen-antibody reaction principle, using Image-Pro Plus software we performed total colorimetry quantification and comparison of immunohistochemical reaction; revealed with diaminobenzidine (IHC-DAB) and FITC fluorescence (IHC-F). As biological model, GPR43 protein was assessed by Immunohistochemestry in inguinal, mesenteric and gonadal (WAT) cd1 mice adipose tissue. In predefined areas of digital images, we evaluated the reaction obtained following the same processing conditions. Results: The GPR43 protein in both IHC-DAB and IHC-F was evidenced in adipocytes membrane and infiltrated immune cells. The average of positive reaction in μm² (total colorimetry x μm²) on a predefined area of 25x10⁴ μm², showed no significant differences in the distinct tissues between the performed methods. Conclusions: In the total colorimetric analysis between the two methodologies IHC-DAB and IHC-F, the resulting quantitative values were similar for all adipose tissue compartments studied.

The mean molecular connectivity indices (MMCI) proposed and used in previous studies are used here in conjunction with the well-known molecular connectivity indices (MCI) to remodel six properties of organic solvents. The MMCI and MCI descriptors of the multilinear relationships for the six properties, obtained with the multilinear least - squares (MLS) procedure, were used to perform the artificial neural network (ANN) computations. The aim is to detect adavantages and underline the limits of the ANN approach that, even if it is able to refine and improve the model, it is somewhat ‘fuzzy’ concerning the stability of the modeling. The MLS procedure is able to replicate the obtaiend results as long as one wishes, a characteristic not shared by the ANN methodology, which, if on one side increases the quality of a description on the other increases also its overfitting. The present study reveals also how ANN methods prefer MCI relatively to MMCI descriptors. Four different types of ANN computations show that (i) MMCI descriptors are preferred with properties with poor number of points. MLS (ii) is to be preferred over ANN statistical results, with some exceptions, when the number of ANN weights is similar to the number of correlation coefficients of MLS. Furthermore, in (iii) some cases MLS modeling quality is quite similar to the modeling quality of ANN computations.

This researcher was invited to give the talk at IWMEDIC 2016, IV International Workshop. Please, follow the link to see the video presentation uploaded (or to be uploaded soon) at the workshop official site: https://sciforum.net/conference/mol2net-02/iwmedic-04