Synthesis of New Class of Alkyl Azarene Pyridinium Zwitterions via Iodine Mediated sp³ C–H Bond Activation

An efficient and conceptually different approach toward C–H bond activation by using iodine mediated sp³ C–H functionalization for the synthesis of alkyl azaarene pyridinium zwitterions is described. This work has the interesting distinction of being the first synthesis of a new class of alkyl azaarene pyridinium zwitterion via transition-metal-free sp³ C–H bond activation of an alkyl azaarene

Base-Controlled Reactions through an Aldol Intermediate Formed between 2‑Oxoaldehydes and Malonate Half Esters

A practical, atom-economical, base-directed, and highly efficient method for the generation of different selective products through a common aldol intermediate of 2-oxoaldehydes and malonate half esters is successfully developed. The addition of a strong basic environment (potassium <i>tert</i>-butoxide) catalyzed the synthesis of stable decarboxylative aldol products (α-hydroxy ketones), while the Doebner modification procedure resulted in decarboxylative elimination to (<i>E</i>)-α,β-unsaturated esters in good yields. The application of this method in one pot and one pot/two steps with azoles helped to develop regioselective α- and β-azolated products in appreciable yields

Regio- and Stereoselective Intermolecular Oxysulfonylation of Alkynes with 1,3-Diketones to Access (<i>Z</i>)‑β-Sulfonated Enethers

The first multicomponent regio- and stereoselective difunctionalization of alkynes via concomitant C–O and C–S bond formation using 1,3-diketones and sodium sulfinate has been developed for the synthesis of various sulfonated enethers. The viability of this strategy is unveiled by gram-scale, various synthetic modifications and late-stage functionalization. This transformation does not require any prefunctionalization, metal catalysts, and oxidants. The present operationally simple, efficient, and sustainable approach provides various functionalized olefins in a one-pot protocol with high Z-selectivity

2‑Oxo-Driven N₂ Elimination Induced Decarbonylative Cyclization Reaction in Benzotriazoles to 6‑Aminophenanthridines

An efficient functional group induced strategy for the synthesis of 6-aminophenanthridines (6AP) has been developed as a result of an in situ generated novel system “CO–CH­(N₁N₂)”. This reaction presents a new mode of N₂ extrusion in benzotriazoles that later result in decarbonylative cyclization to 6AP. This method offers an easier protocol for the synthesis of 6AP from readily available inexpensive substrates

Mesenchymal Stem Cells from Human Extra Ocular Muscle Harbor Neuroectodermal Differentiation Potential

<div><p>Mesenchymal stem cells (MSC) have been proposed as suitable candidates for cell therapy for neurological disorderssince they exhibit good neuronal differentiation capacity. However, for better therapeutic outcomes, it is necessary to isolate MSC from a suitable tissue sourcethat posses high neuronal differentiation. In this context, we isolated MSC from extra ocular muscle (EOM) tissue and tested the <i>in vitro</i> neuronal differentiation potential. In the current study, EOM tissue derived MSC were characterized and compared with bone marrow derived MSC. We found that EOM derived MSC proliferated as a monolayer and showed similarities in morphology, growth properties and cell surface marker expression with bone marrow derived MSC and expressed high levels of NES, OCT4, NANOG and SOX2 in its undifferentiated state. They also expressed embryonic cell surface marker SSEA4 and their intracellular mitochondrial distribution pattern was similar to that of multipotent stem cells. Although EOM derived MSC differentiated readily into adipocytes, osteocytes and chondrocytes, they differentiated more efficiently into neuroectodermal cells. The differentiation into neuroectodermal cellswas confirmed by the expression of neuronal markers NGFR and MAP2B. Thus, EOM derived MSC might be good candidates for stem cell based therapies for treating neurodegenerative diseases.</p></div

Isolation and expansion of EOM-MSC.

<p>(A) EOM-MSC were isolated from freshly resected EOM tissue from the human eye. The tissue was mechanically dissociated and plated in growth media. Phase-contrast microscopic images of (B) EOM-MSC and (C) BM-MSCshowing spindle shaped adherent cells, passage 2. (D) FESEM analysis of <i>in vitro</i> expanded EOM-MSC (passage 3). (E, e’) CD90 expression in <i>in vitro</i> expanded EOM-MSC determined by immunocytochemical staining. Representative photomicrographs are shown. EOM-MSC were from passage 2–4.</p

Gene expression analysis in EOM-MSC.

<p>(A) Expression levels of transcription factors OCT4, NANOG and SOX2 and neuronal lineage related gene NES in EOM-MSC was analysed by real-time PCR and compared with BM-MSC. mRNA expression levels of (B) apoptosis related genes BAD, cIAP1, cIAP2 and (C) other factors HIF1α, FGF2, IL6, TNFα as well as calcium channel related genes ORAI1, STIM1, TRPC1 were determined in EOM-MSC and BM-MSC by real-time PCR. The expression levels of the genes were normalized to GAPDH expression levels in the respective samples. Values are mean±SD, n = 3–5* p<0.05.</p

Neuronal differentiation of EOM-MSC.

<p>(A) Real-time PCR analysis of neuronal specific genes NES, NEUROD1, PAX6 and TUBB3 in EOM-MSC cultured in growth media and neuronal differentiation media for 14 days. The expression levels of the genes were normalized to GAPDH expression levels in the respective samples. Values are Mean±SEM, n = 5, *p<0.05. Immunoflurescence analysis of neuronal differentiated EOM-MSC showing downregulation of CD146 in cells cultured in (B) neuronal media compared to cells grown in (C) normal growth media. (D) CD90 expression was unaffected in EOM-MSC cultured in neuronal media. Neuronal differentiated cells exhibited upregulation of neuronal lineage markers (E) GFAP, (F) NGFR and (G) MAP2B. CD90, CD146 and GFAP staining were performed 9–10 days after the addition of neuronal induction media whereas NGFR and MAP2B staining were performed after 21 days of neuronal induction. Representative microscopic images are shown.</p

EOM derived cells express markers for MSC.

<p>(A) Representative histograms of flow cytometric analysis of EOM-MSC showing cell surface expression for CD13, CD29, CD34, CD44, CD45, CD49A, CD49B, CD49C, CD49D, CD49E, CD49F, CD73, CD90, CD104, CD105, CD140a, CD146, CD200, CD271, HLA I, HLA II, GFAP and MAP2B. Grey histogram represents the isotype control and black line represents the stained sample. EOM-MSC were at passage 2–5. (B) Dot plot showing the flow cytometric analysis of SSEA4 (stage specific embryonic antigen 4) expression in EOM-MSC and (C) fluorescent microscopic picture of EOM-MSC stained with antibody against SSEA4. The arrows indicate spindle shaped cells stained positive for SSEA4. Representative microscopic images are shown, EOM-MSC were from passage 3.</p

Neuronal differentiation of clonal derived EOM-MSC.

<p>(A) Microscopic image of single cell after 24hr of plating to isolate single cell derived colonies. (B) Single cell derived colony 7 days after plating. (C, D) Neuronal differentiation of single cell derived clones of EOM-MSC was determined by staining with NGFR 14 days after neuronal induction Representative microscopic images are shown.</p