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  1. Article: Outcome Predictors of Percutaneous Cholecystostomy As Definitive Versus Bridging Treatment for Acute Cholecystitis.

    Lau, Joshua / Sinha, Surajit

    Cureus

    2023  Volume 15, Issue 12, Page(s) e49962

    Abstract: Introduction Percutaneous cholecystostomy (PC) is a treatment option for patients with acute cholecystitis (AC) who are too unwell, or too morbid for laparoscopic cholecystectomy (LC). Some patients have PC as a definitive treatment, whereas others have ... ...

    Abstract Introduction Percutaneous cholecystostomy (PC) is a treatment option for patients with acute cholecystitis (AC) who are too unwell, or too morbid for laparoscopic cholecystectomy (LC). Some patients have PC as a definitive treatment, whereas others have PC as a bridging treatment prior to LC. The aim of this study is to investigate patient characteristics and mortality among those who received PC as definitive treatment versus bridging treatment. Methods Our study retrospectively reviewed all patients treated with PC for AC from February 2019 to November 2022 at the Torbay and South Devon NHS Foundation Trust, Torquay, England. Fifty patients underwent PC for AC, with 48 patients having follow-up data available for analysis. Of these, 26 patients (54%) only received PC (definitive PC), and 22 patients (46%) later underwent LC (bridging LC). Results In this study, 68.8% of the patients were male, with a mean age of 76 ± 9 years. The overall mean Charlson Comorbidity Index (CCI) score was 4.96 ± 1.12, and the mean American Society of Anesthesiologists (ASA) score was 2.83 ± 0.36. The median PC drain duration was 42 days. Six patients (12.5%) had a recurrence of AC with a mean of 57 days onset after PC insertion. Twelve patients (25%) experienced PC complications: 11 (23%) were minor, involving pain or a dislodged tube, and one (2%) was major, resulting in a subhepatic abscess. The median duration from PC insertion to LC surgery was 50.5 days. The bridging LC cohort had a 30-day and one-year mortality of 0%, while the definitive PC cohort had a 30-day mortality of 30.8% (eight patients) and a one-year mortality of 46.1% (12 patients). The bridging LC cohort compared to the definitive PC cohort had a significantly lower CCI (4.39 vs 5.57, p<0.05), and a significantly lower ASA (2.61 vs 3.04, p<0.05). The one-year survival cohort compared to the 30-day mortality cohort had significantly lower ASA (2.71 vs 3.25 p<0.05), and a non-significantly lower CCI (4.66 vs 5.86 p=0.094). The presence of negative predictive factors of respiratory dysfunction and hyperbilirubinemia had higher 30-day and 90-day mortality rates of 31.3% and 37.5%, compared to their absence of 9.4% and 21.4% respectively. Conclusion Our results demonstrate that PC is a safe procedure with a high success rate and low complications. We showed that PC is an effective treatment option for bridging a select cohort of patients to receive a delayed LC. Furthermore, the data suggests ASA and CCI scoring can be used as clinical adjuncts to assess whether bridging patients from PC to LC is appropriate. Finally, ASA, respiratory dysfunction, and hyperbilirubinemia can be used as significant negative predictors of post-PC mortality.
    Language English
    Publishing date 2023-12-05
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2747273-5
    ISSN 2168-8184
    ISSN 2168-8184
    DOI 10.7759/cureus.49962
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  2. Article ; Online: Synthesis of Self Permeable Antisense PMO Using C5-Guanidino-Functionalized Pyrimidines at the 5'-End Enables Sox2 Downregulation in Triple Negative Breast Cancer Cells.

    Das, Arnab / Gupta, Shalini / Shaw, Pallab / Sinha, Surajit

    Molecular pharmaceutics

    2024  Volume 21, Issue 3, Page(s) 1256–1271

    Abstract: Delivery of macromolecular drugs inside cells has been a huge challenge in the field of oligonucleotide therapeutics for the past few decades. Earliest natural inspirations included the arginine rich stretch of cell permeable HIV-TAT peptide, which led ... ...

    Abstract Delivery of macromolecular drugs inside cells has been a huge challenge in the field of oligonucleotide therapeutics for the past few decades. Earliest natural inspirations included the arginine rich stretch of cell permeable HIV-TAT peptide, which led to the design of several molecular transporters with varying numbers of rigid or flexible guanidinium units with different tethering groups. These transporters have been shown to efficiently deliver phosphorodiamidate morpholino oligonucleotides, which have a neutral backbone and cannot form lipoplexes. In this report, PMO based delivery agents having 3 or 4 guanidinium groups at the C5 position of the nucleobases of cytosine and uracil have been explored, which can be assimilated within the desired stretch of the antisense oligonucleotide. Guanidinium units have been connected by varying the flexibility with either a saturated (propyl) or an unsaturated (propargyl) spacer, which showed different serum dependency along with varied cytoplasmic distribution. The effect of cholesterol conjugation in the delivery agent as well as at the 5'-end of full length PMO in cellular delivery has also been studied. Finally, the efficacy of the delivery has been studied by the PMO mediated downregulation of the stemness marker Sox2 in the triple-negative breast cancer cell line MDA-MB 231. These results have validated the use of this class of delivery agents, which permit
    MeSH term(s) Humans ; Triple Negative Breast Neoplasms/genetics ; Triple Negative Breast Neoplasms/therapy ; Down-Regulation ; Pyrimidines ; Guanidine ; Morpholinos/chemistry ; Oligonucleotides
    Chemical Substances Pyrimidines ; Guanidine (JU58VJ6Y3B) ; Morpholinos ; Oligonucleotides
    Language English
    Publishing date 2024-02-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2138405-8
    ISSN 1543-8392 ; 1543-8384
    ISSN (online) 1543-8392
    ISSN 1543-8384
    DOI 10.1021/acs.molpharmaceut.3c00924
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  3. Article ; Online: Synthesis and Biophysical Properties of Triazole-Incorporated PMOs (TzPMOs): A Convergent, Click Ligation Approach.

    Banerjee, Arpan / Das, Arnab / Ghosh, Atanu / Gupta, Abhishek / Sinha, Surajit

    The Journal of organic chemistry

    2024  Volume 89, Issue 5, Page(s) 2895–2903

    Abstract: The synthesis of phosphorodiamidate morpholino oligonucleotides (PMOs) incorporating single or double triazole rings in the backbone has been achieved via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). The synthetic approach implemented is ... ...

    Abstract The synthesis of phosphorodiamidate morpholino oligonucleotides (PMOs) incorporating single or double triazole rings in the backbone has been achieved via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). The synthetic approach implemented is fundamentally convergent, involving the ligation of a 5'-azide PMO fragment to a 3'-alkyne fragment both in solution and on solid support. To access the 3'-alkyne PMO fragment, we synthesized 3'-
    MeSH term(s) Azides ; Morpholinos ; Alkynes ; Circular Dichroism ; Triazoles
    Chemical Substances Azides ; Morpholinos ; Alkynes ; Triazoles
    Language English
    Publishing date 2024-02-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 123490-0
    ISSN 1520-6904 ; 0022-3263
    ISSN (online) 1520-6904
    ISSN 0022-3263
    DOI 10.1021/acs.joc.3c02242
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: C5-pyrimidine-functionalized morpholino oligonucleotides exhibit differential binding affinity, target specificity and lipophilicity.

    Das, Arnab / Ghosh, Atanu / Sinha, Surajit

    Organic & biomolecular chemistry

    2023  Volume 21, Issue 6, Page(s) 1242–1253

    Abstract: C5-substituted uridine and cytidine morpholino chlorophosphoramidate monomers were synthesized and incorporated into a 12-mer Phosphorodiamidate Morpholino Oligonucleotide (PMO) using semi-automated solid phase synthesis. PMOs with most of the tested ... ...

    Abstract C5-substituted uridine and cytidine morpholino chlorophosphoramidate monomers were synthesized and incorporated into a 12-mer Phosphorodiamidate Morpholino Oligonucleotide (PMO) using semi-automated solid phase synthesis. PMOs with most of the tested pyrimidine C5-substitutions have significantly increased thermal stability when bound to the complementary RNA strand relative to the PMO. They exhibit higher binding with RNA than DNA. CD-spectra show B-type helical conformation of duplexes. HPLC analysis indicates their greater lipophilicity compared to regular PMOs. These chemical modifications have significant potential towards the development of better antisense technologies.
    MeSH term(s) Morpholinos/chemistry ; Oligonucleotides, Antisense/chemistry ; Pyrimidines ; DNA/chemistry ; RNA
    Chemical Substances Morpholinos ; Oligonucleotides, Antisense ; Pyrimidines ; DNA (9007-49-2) ; RNA (63231-63-0)
    Language English
    Publishing date 2023-02-08
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2097583-1
    ISSN 1477-0539 ; 1477-0520
    ISSN (online) 1477-0539
    ISSN 1477-0520
    DOI 10.1039/d2ob01759h
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Synthesis of 5'-Thiol Functionalized Morpholino Oligo-Nucleotide and Subsequent Conjugation with IGT to Improve Delivery and Antisense Efficacy

    Ghosh, Ujjwal / Gupta, Shalini / Sinha, Surajit

    Bioconjugate chemistry

    2022  Volume 34, Issue 1, Page(s) 174–180

    Abstract: Thiol functionalized oligonucleotides are useful intermediates for a wide range of applications including DNA nanobiotechnology field through conjugation with various types of probes and cargos. Due to the limitation of synthetic process, ... ...

    Abstract Thiol functionalized oligonucleotides are useful intermediates for a wide range of applications including DNA nanobiotechnology field through conjugation with various types of probes and cargos. Due to the limitation of synthetic process, phosphorodiamidate morpholino oligonucleotides (PMOs) have not been explored like other oligonucleotides through SH conjugation as mentioned above. In this paper, we report the synthesis of 5'-SH functionalized PMO using a solid support synthesis protocol with an optimized cysteine derived linker so that loading and coupling efficiency of morpholino monomers were effective enough to get a 25-mer 5'-SH functionalized PMO against human Nanog. The PMO with SH functionality was subsequently conjugated with our previously reported Internal Oligo-guanidinium Transporter (IGT) in solution phase to obtain the IGT-PMO conjugate. Interestingly, 5'-conjugated PMO (IGT-PMO) showed 2.5 times better antisense efficacy than 3'-conjugated PMO with IGT (PMO-IGT). 5'-Conjugation enables us to use IGT-PMO for further conjugation at the 3'-
    MeSH term(s) Humans ; Morpholinos ; Guanidine ; Oligonucleotides, Antisense/therapeutic use ; Nucleotides ; Sulfhydryl Compounds
    Chemical Substances Morpholinos ; Guanidine (JU58VJ6Y3B) ; Oligonucleotides, Antisense ; Nucleotides ; Sulfhydryl Compounds
    Language English
    Publishing date 2022-12-20
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1024041-x
    ISSN 1520-4812 ; 1043-1802
    ISSN (online) 1520-4812
    ISSN 1043-1802
    DOI 10.1021/acs.bioconjchem.2c00587
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Guanidinium-Functionalized Flexible Azaproline Transporter for Efficient Intracellular Delivery of Proapoptotic Peptide and PDL1 Antisense Morpholino Oligo in Human Carcinoma Cells

    Gupta, Abhishek / Gupta, Shalini / Das, Ujjal / Sinha, Surajit

    Bioconjugate chemistry

    2022  Volume 33, Issue 5, Page(s) 907–917

    Abstract: Cell-penetrating peptides (CPPs) are structurally diverse sophisticated tools endowed with high arginine content, amphipathicity, and well-adopted suitable secondary structures. Despite its capability of breaching the lipid barriers, CPP has major ... ...

    Abstract Cell-penetrating peptides (CPPs) are structurally diverse sophisticated tools endowed with high arginine content, amphipathicity, and well-adopted suitable secondary structures. Despite its capability of breaching the lipid barriers, CPP has major limitations such as
    MeSH term(s) Animals ; B7-H1 Antigen ; Carcinoma ; Cell-Penetrating Peptides/chemistry ; Cricetinae ; Cricetulus ; Guanidine ; Humans ; Morpholinos
    Chemical Substances B7-H1 Antigen ; Cell-Penetrating Peptides ; Morpholinos ; Guanidine (JU58VJ6Y3B)
    Language English
    Publishing date 2022-04-29
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1024041-x
    ISSN 1520-4812 ; 1043-1802
    ISSN (online) 1520-4812
    ISSN 1043-1802
    DOI 10.1021/acs.bioconjchem.2c00129
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: IGT mediated Nanog siRNA delivery in prostate cancer cells improves chemosensitization of Epirubicin in vitro.

    Gupta, Shalini / Das, Ujjal / Sinha, Surajit

    Bioorganic & medicinal chemistry letters

    2022  Volume 76, Page(s) 129017

    Abstract: Despite the enormous potential of siRNAs to transcriptionally downregulate disease causing proteins in many genetic diseases, efficient delivery and endosomal escape are the two bottlenecks that have resulted in only a handful of FDA approved drugs. In ... ...

    Abstract Despite the enormous potential of siRNAs to transcriptionally downregulate disease causing proteins in many genetic diseases, efficient delivery and endosomal escape are the two bottlenecks that have resulted in only a handful of FDA approved drugs. In this report, we have successfully delivered siRNA against Nanog with the help of pentafluorobenzyl modified Internal Oligo-guanidinium transporter (IGT) that has previously shown promising results in peptide and antisense morpholino delivery. Nanog downregulation in prostate cancer cell line DU145 in serum containing media led to suppression of associated proteins such as KLF4, FAK and cMyc and also enhanced the chemosensitivity of Epirubicin, an anthracycline based drug, in DU145 cells by associated MDR-1 downregulation in vitro. These results show that IGT is a promising candidate for siRNA delivery and its conjugation with stable siRNAs could enhance the chemotherapeutic efficiency of siRNAs alone and in combination with small molecule-based drugs.
    MeSH term(s) Humans ; Male ; Cell Line, Tumor ; Epirubicin/pharmacology ; Guanidine/metabolism ; Morpholinos ; Nanog Homeobox Protein/genetics ; Peptides/administration & dosage ; Prostatic Neoplasms/drug therapy ; Prostatic Neoplasms/genetics ; RNA, Small Interfering/administration & dosage ; Drug Resistance, Neoplasm/genetics ; Organic Cation Transport Proteins
    Chemical Substances Epirubicin (3Z8479ZZ5X) ; Guanidine (JU58VJ6Y3B) ; Morpholinos ; Nanog Homeobox Protein ; NANOG protein, human ; Peptides ; RNA, Small Interfering ; Organic Cation Transport Proteins
    Language English
    Publishing date 2022-10-06
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1063195-1
    ISSN 1464-3405 ; 0960-894X
    ISSN (online) 1464-3405
    ISSN 0960-894X
    DOI 10.1016/j.bmcl.2022.129017
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  8. Article ; Online: Synthesis and Biophysical Studies of High-Affinity Morpholino Oligomers Containing G-Clamp Analogs.

    Das, Arnab / Ghosh, Atanu / Kundu, Jayanta / Egli, Martin / Manoharan, Muthiah / Sinha, Surajit

    The Journal of organic chemistry

    2023  Volume 88, Issue 21, Page(s) 15168–15175

    Abstract: Successful syntheses of chlorophosphoramidate morpholino monomers containing tricyclic cytosine analogs phenoxazine, G-clamp, and ... ...

    Abstract Successful syntheses of chlorophosphoramidate morpholino monomers containing tricyclic cytosine analogs phenoxazine, G-clamp, and G
    MeSH term(s) Morpholinos/chemistry ; Oligonucleotides/chemistry ; DNA/chemistry ; RNA/chemistry ; Nucleic Acid Conformation
    Chemical Substances Morpholinos ; 9-(2-aminoethoxy)phenoxazine ; Oligonucleotides ; DNA (9007-49-2) ; RNA (63231-63-0)
    Language English
    Publishing date 2023-10-16
    Publishing country United States
    Document type Journal Article
    ZDB-ID 123490-0
    ISSN 1520-6904 ; 0022-3263
    ISSN (online) 1520-6904
    ISSN 0022-3263
    DOI 10.1021/acs.joc.3c01658
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Morpholino oligonucleotide-mediated exon skipping for DMD treatment: Past insights, present challenges and future perspectives.

    Gupta, Shalini / Sharma, Swrajit Nath / Kundu, Jayanta / Pattanayak, Sankha / Sinha, Surajit

    Journal of biosciences

    2023  Volume 48

    Abstract: Duchenne muscular dystrophy (DMD) is an X-linked genetic disease primarily affecting boys causing loss of the dystrophin protein, ultimately leading to muscle wastage and death by cardiac or respiratory failure. The genetic mutation involved can be ... ...

    Abstract Duchenne muscular dystrophy (DMD) is an X-linked genetic disease primarily affecting boys causing loss of the dystrophin protein, ultimately leading to muscle wastage and death by cardiac or respiratory failure. The genetic mutation involved can be overcome with antisense oligonucleotides which bind to a pre-mRNA and results in reading frame restoration by exon skipping. Phosphorodiamidate morpholino oligonucleotides (PMOs) are a class of antisense agents with a neutral backbone derived from RNA which can induce effective exon skipping. In this review, the evolution of PMOs in exon skipping therapy for the last two decades has been detailed with the gradual structural and functional advancements. Even though the success rate of PMObased therapy has been high with four FDA approved drugs, several key challenges are yet to overcome, one being the dystrophin restoration in cardiac muscle. The current scenario in further improvement of PMOs has been discussed along with the future perspectives that have the potential to revolutionize the therapeutic benefits in DMD.
    MeSH term(s) Male ; Humans ; Morpholinos/genetics ; Morpholinos/therapeutic use ; Dystrophin/genetics ; Dystrophin/metabolism ; Muscular Dystrophy, Duchenne/genetics ; Muscular Dystrophy, Duchenne/therapy ; Oligonucleotides, Antisense/genetics ; Oligonucleotides, Antisense/therapeutic use ; Exons/genetics
    Chemical Substances Morpholinos ; Dystrophin ; Oligonucleotides, Antisense
    Language English
    Publishing date 2023-10-16
    Publishing country India
    Document type Review ; Journal Article
    ZDB-ID 756157-x
    ISSN 0973-7138 ; 0250-5991
    ISSN (online) 0973-7138
    ISSN 0250-5991
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  10. Article ; Online: Synthesis of Chlorophosphoramidate Monomer Morpholinos and PMOs.

    Kundu, Jayanta / Ghosh, Ujjwal / Ghosh, Atanu / Pattanayak, Sankha / Das, Arnab / Sinha, Surajit

    Current protocols

    2023  Volume 3, Issue 2, Page(s) e686

    Abstract: Phosphorodiamidate morpholino oligonucleotides (PMOs) are a successful class of antisense reagents that efficiently modulate gene expression. Because PMOs do not follow standard phosphoramidite chemistry, optimized synthetic protocols for these compounds ...

    Abstract Phosphorodiamidate morpholino oligonucleotides (PMOs) are a successful class of antisense reagents that efficiently modulate gene expression. Because PMOs do not follow standard phosphoramidite chemistry, optimized synthetic protocols for these compounds are relatively scarce in the literature. This paper presents detailed protocols for synthesizing full-length PMOs using chlorophosphoramidate chemistry by manual solid-phase synthesis. We first describe the synthesis of Fmoc-protected morpholino hydroxyl monomers, and the corresponding chlorophosphoramidate monomers, from commercially available protected ribonucleosides. The new Fmoc chemistry necessitates the use of a milder base, such as N-ethylmorpholine (NEM), and coupling reagent, such as 5-(ethylthio)-1H-tetrazole (ETT), which are also tolerated for acid-sensitive trityl chemistry. These chlorophosphoramidate monomers are then employed for PMO synthesis in a manual solid-phase procedure using four sequential steps. The synthetic cycle for each nucleotide incorporation consists of (a) deblocking of the 3'-N protecting group using an acidic deblocking cocktail for trityl and base deblocking for Fmoc, (b) neutralization, (c) coupling in the presence of ETT and NEM, and (d) capping of the unreacted morpholine ring-amine. The method uses safe, stable, and inexpensive reagents, and the process is expected to be scalable. After full-length PMO synthesis and ammonia-mediated cleavage from the solid support and deprotection, a range of PMOs with different lengths can be obtained conveniently and efficiently with reproducible good yields. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of the novel Fmoc-protected morpholino monomers Basic Protocol 2: Synthesis of the phosphorylating reagent (N,N-dimethylphosphoramic dichloride) required for chlorophosphoramidate monomer synthesis Basic Protocol 3: Synthesis of chlorophosphoramidate monomers of Fmoc-protected morpholino monomers Basic Protocol 4: Solution-phase standardization of dimer and trimer PMO synthesis using Fmoc chemistry Basic Protocol 5: Solid-phase synthesis, purification, and characterization of full-length (25-mer) no-tail PMO using both trityl and Fmoc chemistry.
    MeSH term(s) Morpholinos/chemistry ; Oligonucleotides, Antisense/chemistry
    Chemical Substances Morpholinos ; Oligonucleotides, Antisense
    Language English
    Publishing date 2023-02-19
    Publishing country United States
    Document type Journal Article
    ISSN 2691-1299
    ISSN (online) 2691-1299
    DOI 10.1002/cpz1.686
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