Cross-Cultural Adaptation and Psychometric Validation of the Dutch Version of the Core Outcome Measures Index for the Neck in Patients Undergoing Surgery for Degenerative Disease of the Cervical Spine

Pravesh S. Gadjradj,Timothy C. Chin-See-Chong,Daphne Donk,Paul Depauw,Maurits W. van Tulder,Biswadjiet S. Harhangi 대한척추신경외과학회 2021 Neurospine Vol.18 No.4

Objective: To perform the psychometric validation of the Dutch version of the Core Outcome Measures Index (COMI) for the neck. Methods: A total of 178 patients, who had an indication for surgery due to degenerative cervical spinal disease, were enrolled in the study. They filled in a baseline booklet containing the Dutch version of the COMI-neck, Likert-scales for neck and arm/shoulder pain, the Neck Disability Index (NDI), the EuroQol-5 dimensions (EQ-5D) and the 12-item Short Form health survey (SF-12). Aside from analyzing construct validity using the Spearman correlation test, test-retest reliability, and responsiveness at 3 months were assessed using the intraclass correlation coefficient (ICC) and the receiver-operating characteristic (ROC) curve, respectively. Results: The COMI-neck showed good acceptability with missing data ranging from 0% to 4.5% and some floor/ceiling effects for 3 of the domains at baseline. The COMI-summary score showed good to very good correlation with the EQ5D (ρ=-0.43), the physical component summary of the SF-12 (ρ=-0.47) and the NDI (ρ=0.73). Individual domains showed correlations of -0.28 to 0.85 with the reference questionnaires. Test-retest reliability analysis showed an ICC of 0.91 with a minimal detectable change of 1.7. Responsiveness analysis of the COMI-neck showed an area under 0.79 under the ROC-curve. The standardized response mean for a good outcome was 1.24 and for a poor outcome 0.37. Conclusion: The current study shows that the Dutch version of the COMI-neck is a valid, reliable and responsive Patient-Reported Outcome Measure, among patients undergoing surgery for degenerative cervical spinal disorders.

Surgeons Learning Curve of Transforaminal Endoscopic Discectomy for Sciatica

Pravesh S. Gadjradj,Arnold Vreeling,Paul R. Depauw,Pieter J. Schutte,Biswadjiet S. Harhangi,PTED-Study Group 대한척추신경외과학회 2022 Neurospine Vol.19 No.3

Objective: Full-endoscopic spine surgery is gaining interest as a less-invasive alternative to treat sciatica caused by a lumbar disc herniation. Concerns, however, exist with the learning curve as percutaneous transforaminal endoscopic discectomy (PTED) appears to be more difficult to be performed compared to other techniques. In this study, the clinical outcomes during and after the learning curve are presented of 3 surgeons naïve to PTED. Methods: In the first phase of a randomized controlled, noninferiority trial comparing PTED with microdiscectomy, 3 surgeons were trained in the PTED-procedure by a senior surgeon. After performing up to 20 cases under supervision, they started performing PTED on their own. Results of the early cases were compared to the later cases (>20). Furthermore, complications and reoperations were compared. Finally, differences in clinical outcomes between surgeons were compared. Results: At 12 months of follow-up, 87% of the patients had follow-up data available. In general, there were no significant differences in patient-reported outcomes between the early and later PTED cases. Furthermore, outcomes of the early PTED cases were comparable to the outcomes of microdiscectomy, while the later PTED cases had small, but more favorable outcomes compared to microdiscectomy. Two learning curve surgeons had substantially higher rates of reoperations within 1 year, compared to the senior surgeon or the microdiscectomy group. Duration of surgery was also longer for all learning curve surgeons. Finally, when comparing clinical outcomes of patients undergoing PTED versus microdiscectomy, there appears to be some statistically significant differences in outcomes compared between the senior and 3 learning curve surgeons. Conclusion: PTED appears to be safe to be adopted by surgeons naïve to the procedure when they are initially supervised by an experienced senior surgeon. Duration of surgery and risk of repeated surgery are increased during the learning curve, but patient-reported outcomes of the early PTED cases are similar to the outcomes of later PTED cases, and similar to the outcomes of microdiscectomy cases. This study underlines the need for an experienced mentor for surgeons to safely adopt PTED.

Assessing the Learning Process of Transforaminal Endoscopic Discectomy for Sciatica

Pravesh Shankar Gadjradj,Pieter Schutte,Arnold Vreeling,Paul Depauw,Biswadjiet S. Harhangi 대한척추신경외과학회 2022 Neurospine Vol.19 No.3

Objective: Percutaneous transforaminal endoscopic discectomy (PTED) is gaining popularity by both surgeons and patients as a less invasive treatment option for sciatica. Concerns, however, exist for its learning curve. No previous study has assessed the learning process of PTED. Hereby we present the learning process of 3 surgeons learning PTED. Methods: This analysis was conducted alongside a multicenter randomized controlled trial. After attending a cadaveric workshop, 3 spine-dedicated surgeons started performing PTED, initially under the supervision of a senior surgeon. After each 5 cases, and up to case 20, the learning process was evaluated using the validated questionnaires (objective structured assessment of technical skills [OSATS], global operative assessment of laparoscopic skills [GOALS]) and a 10-step checklist specifically developed for PTED. Results: In total, 3 learning curve surgeons performed a total of 161 cases. Based on selfassessment, surgeons improved mostly in the domains “time and motion,” “respect for tissue,” and “knowledge and handling of instruments.” Learning curve surgeons were more able to detect differences in performances on the OSATS than the senior surgeon. Based on the GOALS, the biggest improvements could be seen in “depth-perception” and “autonomy.” Based on the 10-item specific checklist, all surgeons performed all 10 steps by case 10, while only 1 surgeon performed all steps adequately by case 15. Conclusion: Based on these study results, PTED appears to be successfully adopted stepwise by 3 spine-dedicated surgeons. From 15 cases on, most steps are performed adequately. However, more cases might be necessary to achieve good clinical results. Validated tools are needed to determine the cutoff when a surgeon should be able to perform PTED independently.

Cloning, Purification, and Characterization of Recombinant Human Extracellular Superoxide Dismutase in SF9 Insect Cells

Shrestha, Pravesh,Yun, Ji-Hye,Kim, Woo Taek,Kim, Tae-Yoon,Lee, Weontae Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.3

A balance between production and degradation of reactive oxygen species (ROS) is critical for maintaining cellular homeostasis. Increased levels of ROS during oxidative stress are associated with disease conditions. Antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD), in the extracellular matrix (ECM) neutralize the toxicity of superoxide. Recent studies have emphasized the importance of EC-SOD in protecting the brain, lungs, and other tissues from oxidative stress. Therefore, EC-SOD would be an excellent therapeutic drug for treatment of diseases caused by oxidative stress. We cloned both the full length (residues 1-240) and truncated (residues 19-240) forms of human EC-SOD (hEC-SOD) into the donor plasmid pFastBacHTb. After transposition, the bacmid was transfected into the Sf9-baculovirus expression system and the expressed hEC-SOD purified using FLAG-tag. Western blot analysis revealed that hEC-SOD is present both as a monomer (33 kDa) and a dimer (66 kDa), as detected by the FLAG antibody. A water-soluble tetrazolium (WST-1) assay showed that both full length and truncated hEC-SOD proteins were enzymatically active. We showed that a potent superoxide dismutase inhibitor, diethyldithiocarbamate (DDC), inhibits hEC-SOD activity.

Expression, Purification and NMR studies of SH3YL1 SH3 domain

Shrestha, Pravesh,Yun, Ji-Hye,Lee, Weon-Tae Korean Magnetic Resonance Society 2010 Journal of the Korean Magnetic Resonance Society Vol.14 No.2

SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of $\beta$-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.

NMR uncovers direct interaction between human NEDD4-1 and p34^SEI−1

Shrestha, Pravesh,Yun, Ji-Hye,Ko, Yoon-Joo,Yeon, Kyu Jeong,Kim, Dooseop,Lee, Heejong,Jin, Dong-Hoon,Nam, Ki-Yup,Yoo, Hye Dong,Lee, Weontae Academic Press 2017 Biochemical and biophysical research communication Vol. No.

<P><B>Abstract</B></P> <P>PTEN, an important tumor suppressor and a key regulator of the PI3K/AKT signaling pathway, is often deleted/mutated in different types of cancer. The E3 ubiquitin ligase NEDD4-1 catalyzes the polyubiquitination of PTEN, thereby acting as a negative regulator of PTEN. Stability of NEDD4-1, in turn, is tightly controlled by a 34 kDa oncoprotein, p34<SUP>SEI−1</SUP> and it regulates PTEN degradation and activates PI3K/AKT pathway, resulting in cancer metastasis. p34<SUP>SEI−1</SUP> affects not only the expression of <I>NEDD4-1</I> during transcription and translation but also the subcellular localization of PTEN. This emphasizes the need to understand, at molecular level, the interaction between NEDD4-1 and p34<SUP>SEI−1</SUP>. A recent study showed that NEDD4-1 interacts with p34<SUP>SEI−1</SUP> via its WWI domain. However, a detailed interaction for molecular level is yet unknown. We report that the WW1 domain of NEDD4-1 recognizes the SERTA domain containing the proline rich region (PRR motif) in p34<SUP>SEI−1</SUP>. TALOS analysis based on NMR data confirms three conserved β-sheets in NEDD4-1 WW1 and the central β-sheet of NEDD4-1 WW1 plays a role for protein stability by the backbone dynamics experiments. NMR titration data revealed the binding site for p34<SUP>SEI−1</SUP> with NEDD4-1. Our data will provide insights into the molecular mechanism of NEDD4-1 and p34<SUP>SEI−1</SUP> interaction, which will be directly used for drug design which inhibits the molecular interaction involved in different cancer signaling.</P> <P><B>Highlights</B></P> <P> <UL> <LI> NEDD4-1 recognizes SERTA domain containing proline rich region of p34<SUP>SEI−1</SUP>. </LI> <LI> TALOS analysis based on NMR data confirms three conserved β-sheets in NEDD4-1 WW1. </LI> <LI> The central β-sheet of NEDD4-1 WW1 provides protein stability as indicated by the backbone dynamics data. </LI> <LI> Residues of NEDD4-1 responsible for direct interaction with p34<SUP>SEI−1</SUP> are identified by NMR titration experiments. </LI> <LI> Our data will be of essence in designing inhibitors for NEDD4-1/p34<SUP>SEI−1</SUP> interaction. </LI> </UL> </P>

C-terminal tail of NADPH oxidase organizer 1 (Noxo1) mediates interaction with NADPH oxidase activator (Noxa1) in the NOX1 complex

Shrestha, Pravesh,Yun, Ji-hye,Ko, Yoon-Joo,Kim, Myeongkyu,Bae, Yun Soo,Lee, Weontae Academic Press 2017 Biochemical and biophysical research communication Vol. No.

<P><B>Abstract</B></P> <P>NOX1 (NADPH oxidase) similar to phagocyte NADPH oxidase, is expressed mainly in the colon epithelium and it is responsible for host defense against microbial infections by generating ROS (reactive oxygen species). NOX1 is activated by two regulatory cytosolic proteins that form a hetero-dimer, Noxo1 (NOX organizer 1) and Noxa1 (NOX activator 1). The interaction between Noxa1 and Noxo1 is critical for activating NOX1. However no structural studies for interaction between Noxa1 and Noxo1 has not been reported till date. Here, we studied the inter-molecular interaction between the SH3 domain of Noxa1 and Noxo1 using pull-down assay and NMR spectroscopy. <SUP>15</SUP>N/<SUP>13</SUP>C-labeled SH3 domain of Noxa1 has been purified for hetero-nuclear NMR experiments (HNCACB, CBCACONH, HNCA, HNCO, and HSQC). TALOS analysis using backbone assignment data of the Noxa1 SH3 domain showed that the structure primarily consists of β-sheets. Data from pull-down assay between the Noxo1 and Noxa1 showed that the SH3 domains (Noxa1) is responsible for interaction with Noxo1 C-terminal tail harboring proline rich region (PRR). The concentration-dependent titration of the Noxo1 C-terminal tail to Noxa1 shows that Noxo1 particularly in the RT loop: Q407*, H408, S409, A412*, G414*, E416, D417, L418, and F420; n-Src loop: C430, E431*, V432*, A435, W436, and L437; and terminal region: I447; F448*, F452* and V454 interact with Noxa1. Our results will provide a detailed understanding for interaction between Noxa1 and Noxo1 at the molecular level, providing insights into their cytoplasmic activity-mediated functioning as well as regulatory role of C-terminal tail of Noxo1 in the NOX1 complex.</P> <P><B>Highlights</B></P> <P> <UL> <LI> C-terminal of Noxo1 harboring PRR motif is important for Noxa1 interaction. </LI> <LI> Noxa1 SH3 binds C-terminal region of Noxo1with K<SUB>d</SUB> of 8.2 ± 1.41 μM. </LI> <LI> Data from NMR experiments determined secondary structures of Noxa1 SH3. </LI> <LI> Noxa1 interacts with PRR region of Noxo1, however, binding affinity is weaker than C-terminal tail of Noxo1. </LI> <LI> Our results provide insights into regulatory role of C-terminal tail of Noxo1 in the NOX1 complex. </LI> </UL> </P>

Patterns of Impaired Neurocognitive Performance on the Global Neuropsychological Assessment, and Their Brain Structural Correlates in Recent-onset and Chronic Schizophrenia

Vineeth Mohan(Vineeth Mohan ),Pravesh Parekh(Pravesh Parekh ),Ammu Lukose(Ammu Lukose ),Sydney Moirangthem(Sydney Moirangthem ),Jitender Saini(Jitender Saini ),David J. Schretlen(David J. Schretlen ) 대한정신약물학회 2023 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.21 No.2

Objective: Schizophrenia is associated with impairment in multiple cognitive domains. There is a paucity of research on the effect of prolonged illness duration (≥ 15 years) on cognitive performance along multiple domains. In this pilot study, we used the Global Neuropsychological Assessment (GNA), a brief cognitive battery, to explore the patterns of cognitive impairment in recent-onset (≤ 2 years) compared to chronic schizophrenia (≥ 15 years), and correlate cognitive performance with brain morphometry in patients and healthy adults. Methods: We assessed cognitive performance in patients with recent-onset (n = 17, illness duration ≤ 2 years) and chronic schizophrenia (n = 14, duration ≥ 15 years), and healthy adults (n = 16) using the GNA and examined correlations between cognitive scores and gray matter volumes computed from T1-weighted magnetic resonance imaging images. Results: We observed cognitive deficits affecting multiple domains in the schizophrenia samples. Selectively greater impairment of perceptual comparison speed was found in adults with chronic schizophrenia (p = 0.009, η2 partial = 0.25). In the full sample (n = 47), perceptual comparison speed correlated significantly with gray matter volumes in the anterior and medial temporal lobes (TFCE, FWE p < 0.01). Conclusion: Along with generalized deficit across multiple cognitive domains, selectively greater impairment of perceptual comparison speed appears to characterize chronic schizophrenia. This pattern might indicate an accelerated or premature cognitive aging. Anterior-medial temporal gray matter volumes especially of the left hemisphere might underlie the impairment noted in this domain in schizophrenia.

Cloning, Purification, and Characterization of Recombinant Human Extracellular Superoxide Dismutase in SF9 Insect Cells

WeontaeLee,Pravesh Shrestha,Ji-Hye Yun,Woo Taek Kim,Tae-Yoon Kim 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.3

A balance between production and degradation of reactive oxygen species (ROS) is critical for maintaining cellular homeostasis. Increased levels of ROS during oxidative stress are associated with disease conditions. Antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD), in the extracellular matrix (ECM) neutralize the toxicity of superoxide. Recent studies have emphasized the importance of EC-SOD in protecting the brain, lungs, and other tissues from oxidative stress. Therefore, EC-SOD would be an excellent therapeutic drug for treatment of diseases caused by oxidative stress. We cloned both the full length (residues 1-240) and truncated (residues 19-240) forms of human EC-SOD (hEC-SOD) into the donor plasmid pFastBacHTb. After transposition, the bacmid was transfected into the Sf9-baculovirus expression system and the expressed hEC-SOD purified using FLAG-tag. Western blot analysis revealed that hEC-SOD is present both as a monomer (33 kDa) and a dimer (66 kDa), as detected by the FLAG antibody. A water-soluble tetrazolium (WST-1) assay showed that both full length and truncated hEC-SOD proteins were enzymatically active. We showed that a potent superoxide dismutase inhibitor, diethyldithio-carbamate (DDC), inhibits hEC-SOD activity.

Expression, Purification and NMR studies of SH3YL1 SH3 domain

이원태,Pravesh Shrestha,윤지혜 한국자기공명학회 2010 Journal of the Korean Magnetic Resonance Society Vol.14 No.2

SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of β-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.