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1.
J Maxillofac Oral Surg ; 23(1): 16-22, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38312965

ABSTRACT

Background: Head and neck cancer is the most common cancer around the globe, following lung cancer and breast cancer. Treatment at advanced stages of head and neck cancer is usually followed intense surgical procedures, which leads to mutilation among patients. Mutilation imparts a sense of disgrace and causes a feeling of shame and stigma in the patient. The feeling of shame and stigma persists over time and affects the overall long-term survival of patients by deteriorating their quality of life. Objectives: Since shame and stigma is an important psychological domain of head and neck cancer, the present article aims toward evaluating the studies published so far for the assessment of shame and stigma in head and neck cancer and highlighting the lacunae in the existing research designs. The present study also aims to design a checklist that could be followed while developing, translating, or validating a psychometric instrument that aims to measure shame and stigma in head and neck cancer. Methods: In the present metanalysis, all articles published in the past years on shame and stigma in head and neck cancer was compiled using a predefined data extraction matrix. The available literature was compiled for major objectives of the study, the sample size used, major findings, and critical lacunae that need to be addressed. Results: Shame and stigma is a very important domain of psychological well-being in head and neck cancer patients, which yet not appropriately addressed and further need to be researched. Conclusion: Future studies could be based on the lacunae highlighted in the existing literature, and the prescribed methodology checklist could be taken into consideration while conducting further studies involving developing, translating, or validating a psychometric instrument related to shame and stigma in the head and neck cancer.

2.
Indian J Otolaryngol Head Neck Surg ; 75(2): 1266-1270, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37274986

ABSTRACT

Background: Self-reported measures are the questionnaire-based instrument that are routinely used in the clinical scenario to assess psychological health. Technically, the self-reported measure should be administrated by the patients themselves but due to the complexity of tools and illiteracy among patients, clinicians often tend to interview the patients. Objective: Present article aims to compare the accuracy of a self-reported measure in the assessment of the psychological health of a patient when the instrument is self-administrated by the patient and when administrated by the clinician or researcher. Methods: We have recruited 43 patients of oral cancer in the study who have a tumor in the buccal mucosa region. The Hindi version of the shame and stigma scale was used to analyse the shame and stigma in patients. The questionnaire was first provided to the patient for the self-administration and after that clinician administrated the questionnaire to the patient by keeping the clinician blinded to the patient self-administrated responses. Results: There was no significant difference in the global mean score and mean score of various subdomains of shame and stigma scale in the self-administered and clinician-administered mode of interview. However, the clinician-administered mode could provide more accurate measures as it helps the patient towards a better understanding of questions. Conclusion: It is recommended that the newly developed or translated self-reported measure should be tested for both patient administrated and clinician administrated compatibility. Questionnaires could be administrated by the clinician in the case when the patient is illiterate or in the case when the patient does not understand the language of the instrument.

3.
Int J Mol Sci ; 24(2)2023 Jan 09.
Article in English | MEDLINE | ID: mdl-36674796

ABSTRACT

Staphylococcus aureus protein A (SpA) is an IgG Fc-binding virulence factor that is widely used in antibody purification and as a scaffold to develop affinity molecules. A cyclized SpA Z domain could offer exopeptidase resistance, reduced chromatographic ligand leaching after single-site endopeptidase cleavage, and enhanced IgG binding properties by preorganization, potentially reducing conformational entropy loss upon binding. In this work, a Z domain trimer (Z3) was cyclized using protein intein splicing. Interactions of cyclic and linear Z3 with human IgG1 were characterized by differential scanning fluorimetry (DSF), surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC). DSF showed a 5 ℃ increase in IgG1 melting temperature when bound by each Z3 variant. SPR showed the dissociation constants of linear and cyclized Z3 with IgG1 to be 2.9 nM and 3.3 nM, respectively. ITC gave association enthalpies for linear and cyclic Z3 with IgG1 of -33.0 kcal/mol and -32.7 kcal/mol, and -T∆S of association 21.2 kcal/mol and 21.6 kcal/mol, respectively. The compact cyclic Z3 protein contains 2 functional binding sites and exhibits carboxypeptidase Y-resistance. The results suggest cyclization as a potential approach toward more stable SpA-based affinity ligands, and this analysis may advance our understanding of protein engineering for ligand and drug development.


Subject(s)
Inteins , Staphylococcus aureus , Humans , Inteins/genetics , Ligands , Thermodynamics , Immunoglobulin G , Calorimetry/methods , Protein Binding
4.
Biotechnol Bioeng ; 120(2): 482-490, 2023 02.
Article in English | MEDLINE | ID: mdl-36225160

ABSTRACT

In the manufacture of therapeutic monoclonal antibodies, the clarified cell culture fluid (CCF) is typically loaded onto an initial protein A affinity capture column. Imperfect mass transfer and loading to maximum capacity can risk antibody breakthrough and loss of valuable product, but conservative underloading wastes expensive protein A resin. In addition, the effects of column fouling and ligand degradation require the frequent optimization of immunoglobulin G (IgG) loading to avoid wastage. Continuous real-time monitoring of IgG flowthrough is of great interest, therefore. We previously developed a fluorescence-based monitoring technology that allows batch mix-and-read mAb detection in the CCF. Here, we report the use of reporters immobilized on cyanogenbromide-activated Sepharose 4B resin for continuous detection of IgG in column breakthrough. The column effluent is continuously contacted with immobilized fluorescein-labeled Fc-binding ligands in a small monitoring column to produce an immediately-detectable change in fluorescence intensity. The technology allows rapid and reliable monitoring of IgG in a flowing stream of clarified CCF emerging from a protein A column, without prior sample preparation. We observed a significant change in fluorescence intensity at 0.5 g/L human IgG, sufficient to detect a 5% breakthrough of a 10 g/L load, within 18 s at a flow rate of 0.5 ml/min. The current small-scale technology is suitable for use in process development, but the chemistry should be readily adaptable to larger scale applications using fiber-optic sensors, and continuous IgG monitoring could be applicable in a variety of upstream and downstream process settings.


Subject(s)
Antibodies, Monoclonal , Immunoglobulin G , Humans , Chromatography, Affinity , Staphylococcal Protein A , Ligands , Coloring Agents
5.
Anal Biochem ; 660: 114929, 2023 01 01.
Article in English | MEDLINE | ID: mdl-36270332

ABSTRACT

Detection and quantification of antibodies, especially immunoglobulin G (IgG), is a cornerstone of ELISAs, many diagnostics, and the development of antibody-based drugs. Current state-of-the-art immunoassay techniques for antibody detection require species-specific secondary antibodies and carefully-controlled bioconjugations. Poor conjugation efficiency degrades assay performance and increases the risk of clinical false positives due to non-specific binding. We developed a generic, highly-sensitive platform for IgG quantification by fusing the IgG-Fc binding Z domain of Staphylococcal Protein A with the ultrabright bioluminescence reporter Nanoluc-luciferase (Nluc). We demonstrated the application of this fusion protein in a sandwich IgG detection immunoassay using surface-bound antigens to capture target IgG and protein A-Nanoluc fusion as the detector. We optimized the platform's sensitivity by incorporating multiple repeats of the Z domain into the fusion protein constructs. Using rabbit and mouse anti-SARS-CoV-2 Nucleoprotein IgGs as model analytes, we performed ELISAs in two different formats, either with SARS-CoV-2 Nucleoprotein as the capture antigen or with polyclonal chicken IgY as the capture antibody. Using standard laboratory equipment, the platform enabled the quantitation of antibody analytes at concentrations as low as 10 pg/mL (67 fM).


Subject(s)
COVID-19 , Immunoglobulin G , Mice , Rabbits , Animals , Staphylococcal Protein A , SARS-CoV-2 , Antibodies, Viral , Immunoassay/methods , Nucleoproteins , Sensitivity and Specificity
6.
Front Public Health ; 10: 843134, 2022.
Article in English | MEDLINE | ID: mdl-35769774

ABSTRACT

Aim: Common Yoga Protocol (CYP) is a standardized yoga protocol authored by experts from all over the world under the aegis of the Ministry of AYUSH, Ayurveda, Yoga and Naturopathy, Unani, Siddha, Sowa Rigpa and Homeopathy (AYUSH). The potential of CYP can be determined as a cost-effective lifestyle modification to prevent the risk of developing cardiovascular diseases (CVD). Methods: In this prospective trial, we compared the effect of CYP at baseline and after 1 month. A total of 374 yoga-naïve participants performed CYP under the supervision of experienced trainers. Physiological [body mass index (BMI), blood pressure, percent oxygen saturation], biochemical (fasting blood glucose and lipid profile), and neurocognitive parameters were measured before and after the intervention. Results: At day 30 of yoga practice, serum levels of low-density lipoprotein (LDL), total cholesterol (TC), and high-density lipoprotein (HDL) were found significantly improved as compared to the baseline levels observed at the time of enrollment. Similarly, the lipid profile was also obtained from experienced trainers and found to be significantly different from those of yoga-naïve volunteers. When the intervention was compared between the healthy yoga-naïve participants with yoga-naïve participants suffering from medical issues, it was found that cholesterol profile improved significantly in the healthy-naive group as compared to the diseased group (hypertension, diabetes, underwent surgery, and CVD). Conclusion: These results highlight the need for further research to better understand the effects of yoga on the primary prevention of CVD.


Subject(s)
Cardiovascular Diseases , Yoga , Cardiovascular Diseases/prevention & control , Cholesterol , Humans , Life Style , Prospective Studies
7.
J Maxillofac Oral Surg ; 21(2): 326-331, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35712431

ABSTRACT

Background: Incidence of oral cancer have been increased aggressively in recent years, and many advanced surgical techniques have developed to facilitate the treatment of disease while preserving the patient's quality of life (QoL). Subjects and Methods: The present study aims toward the evaluation of patient's satisfaction toward the current treatment procedure for oral cancer by assessing the QoL preoperatively and postoperatively. In the study period of four years, we had operated 150 patients with oral cancer (Stage III and IVa) by surgical excision and flap reconstruction, and QoL was assessed using the WHO-QoL BREF questionnaire. Results: We found significant improvement after surgery in various aspects of QoL including the physical health, social health, psychological health and environmental health. Conclusions: The present study is evidence of patient satisfaction toward the current treatment protocols  of oral cancer. However, advanced surgical techniques that can enhance the rate of functional rehabilitation can have paramount importance in improving the patient's QoL.

8.
Indian J Otolaryngol Head Neck Surg ; 74(Suppl 2): 2912-2916, 2022 Oct.
Article in English | MEDLINE | ID: mdl-33907679

ABSTRACT

Social stigma is the spoilation of the social image of an individual, which leads to the social disapproval of the individual by the community. Patients of many diseases like HIV, deafness, and reproductive disorders often face social disproval. Head & neck cancer survivors perceived stigma due to the mutilation that occurred after surgical treatment procedures. The novel coronavirus is a recently emerged zoonotic viral agent that affects the respiratory system of humans. Following its outbreak from the Wuhan city of China, the COVID-19 spreaded fiercely around the globe, forming a pandemic. Since COVID-19 is a contagious disease with no available treatment, social distancing is considered as the best strategy to prevent the geometric spread of infection. With the social distancing model, the head & neck cancer survivors along with the various other stakeholders perceived stigma being a high-risk group for COVID-19 infection. As the pool of caregivers is diminished due to pandemic, head & neck cancer survivors face increased social isolation and perceived stigma in asking for help from relatives. At the time of the pandemic, social support is critical to fighting against the disease. Social distancing should be maintained along with communication with the patients through calls, text, and online social platforms. It is not wise to stigmatize disease as, in that case, patients who are infected with the disease will try to hide it and avoid seeking medical care. With the promotion of social distancing, it is crucial to convey awareness regarding not to stigmatize the disease.

9.
Indian J Otolaryngol Head Neck Surg ; 74(Suppl 3): 5990-5993, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36742652

ABSTRACT

Cancer in the head and neck region is among the most common cancer around the world, the incidence of which keep increasing in past years. Treatment of disease is usually done by the surgical excision which often leads to some degree of facial disfigurements which cause mutilation in patients. Mutilation imparts the feeling of stigma in patients, and patients usually tend to hide facial disfigurements using additional clothing. As a prevention strategy, awareness regarding the disease conveys to the mass population via media commercials. Media commercials which highlight the adverse outcomes of cancer are found to target the stigmatized perspective of disease. On the brighter side, more stigmatized is the patient image in the commercials, more motivation it will create in masses to avoid risk factors like tobacco, smoking and alcohol. But on the darker side, stigmatized commercials create a social environment in which people tend to maintain social distance to cancer patients, and patients have to bear social disapproval by society for their whole life. It reduces the self-esteem and quality of life of patients which affects their overall survival. In the present article, we review the status of stigma in head and neck cancer patients, tools that are available for assessment of stigma, and effects of the stigmatized media commercials on the patient's self-esteem. The present article represents the accurate picture of the problem and highlights the policies which could be employed to balance the paradox of stigmatized media commercials and a healthy social environment for cancer patients.

10.
MAbs ; 13(1): 1980178, 2021.
Article in English | MEDLINE | ID: mdl-34662534

ABSTRACT

Antibodies and Fc fusion proteins are a rapidly growing class of pharmaceuticals. Cell culture and purification process development and operation require frequent measurement of product concentrations, commonly by complex enzyme-linked immunosorbent assay and high-performance liquid chromatography methods. Here we report a fast (<30 s), and simple antibody Fc assay based on mix-and-read reporting by fluorescence emission. A soluble fluorescein-labeled Fc-affinity reporter produced by standard peptide synthesis is mixed with an Fc-containing sample to produce an immediate shift in both fluorescence polarization and intensity, compatible with on- and at-line measurements and microbioreactor monitoring. We observed significant shifts in fluorescence intensity in Chinese hamster ovary cell culture fluid spiked with IgG and detected an adalimumab biosimilar down to 100 ng/mL (10-4 g/L), despite the interferents in the complex sample matrix. Neither the fluorescence polarization nor the fluorescence intensity assay is significantly affected by the addition of clarified lysate of 2 million CHO-k1 cells/mL, suggesting applicability even to cultures of low viability. Biochemical and molecular docking approaches suggest that the fluorescence intensity enhancement is caused by changes in the fluorophore's local microenvironment upon binding to IgG Fc, especially by interactions with Fc His433.Abbreviations: CCF: Cell Culture Fluid; CHO: Chinese Hamster Ovary cells; ELISA: Enzyme Linked Immunosorbent Assay; Fc: Fragment Crystallizable of antibody; HPLC: High-Performance Liquid Chromatography; HPßCD: hydroxypropyl-ß-cyclodextrin; IgG: ImmunoglobulinG; mAb: Monoclonal Antibody; PBS: Phosphate-Buffered Saline; PDB: Protein Data Bank; SpA: Staphylococcal protein A; SpG: Staphylococcal protein G.


Subject(s)
Immunoglobulin Fc Fragments , Staphylococcal Protein A , Animals , CHO Cells , Cricetinae , Cricetulus , Immunoglobulin Fc Fragments/chemistry , Molecular Docking Simulation
11.
J Cancer Res Ther ; 16(3): 517-520, 2020.
Article in English | MEDLINE | ID: mdl-32719260

ABSTRACT

AIM OF THE STUDY: Both matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) is involved in degradation of extracellular matrix and found to stimulate invasion and metastasis in cancer patients. However, studies on the stage-specific expression of MMPs at different stages of larynx carcinoma are still lacking. In the present study, we compare the expression level of MMP-2 and MMP-9 at different stages of laryngeal carcinoma. MATERIAL AND METHODS: Tumor tissues samples were taken from larynx cancer patients by deep biopsy during direct laryngoscopy. Gene expression for MMP-2 and MMP-9 was analyzed using RT-PCR. RESULTS: Significantly high expression of MMP-2 was observed compared to the MMP-9 at stage IV compared to the less advanced stages of the disease. CONCLUSION: Present study concluded that the MMP-2 expressed with a greater magnitude as compared to the MMP-9 in advance stages of laryngeal carcinoma.


Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/pathology , Gene Expression Regulation, Neoplastic , Laryngeal Neoplasms/pathology , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 9/genetics , Adult , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/surgery , Humans , Laryngeal Neoplasms/genetics , Laryngeal Neoplasms/surgery , Middle Aged , Neoplasm Staging , RNA, Messenger/genetics , Young Adult
12.
J Indian Assoc Pediatr Surg ; 25(3): 155-162, 2020.
Article in English | MEDLINE | ID: mdl-32581443

ABSTRACT

AIMS: We are reporting single-institution's experience regarding the role of conservative management in 38 cases of minor and major anastomotic leaks [AL] occurring after primary surgery of esophageal atresia [EA] with tracheo-esophageal fistula [TEF] during last 17 years between 2000 and 2017. In this retrospective review, we are sharing our experience and protocol of management of AL with more emphasis to evaluate: (a) role of conservative treatment in major AL (b) role of extra-pleural approach in enhancing the success rate in conservative treatment in major AL (c) to define the criteria for major & minor leaks and (d) to evaluate the role of ventilation in primary EA surgery to control AL. METHODS: All these cases were operated through extra-pleural approach and out of total 203 cases, 38[18.7%] developed anastomotic leaks. In 29 of the 38 cases [14.3%], leak was minor and in 9 cases [4.4%] the leak was a major one. All these cases of leaks were managed conservatively. RESULTS: All cases of major and minor leaks showed spontaneous healing except one case of minor leak that died before healing due to major cardiac anomaly. For minor leaks, average healing time was 9.5 days while for major leaks it was 17.4 days. Overall mortality was 14.8% and there was no mortality directly attributable to major or minor leak. During follow up, the incidence of stricture was 40% in cases having anastomotic leaks, while in cases without a leak, the incidence of stricture was 23.3%. These all cases of stricture responded to regular dilatations. CONCLUSION: We believe in cases of major AL, where primary repair is done by EP approach, a conservative treatment should be the treatment of choice. With this conservative approach of management of major AL, we not only save the native esophagus, the best conduit, but there is also less morbidity and mortality.

13.
Ann Neurosci ; 26(1): 21-24, 2019 Jan.
Article in English | MEDLINE | ID: mdl-31975768

ABSTRACT

BACKGROUND: Attempts for Guinness world record have continued worldwide but these attempts were rarely aimed to promote public health. Diabetes is one of the rapidly growing lifestyle disorders in India which requires awareness reinforcements among the local population. In recent studies, Yoga has proved to be useful in lifestyle modification and Diabetes management. However, most individuals from rural and urban localities in the country are unaware of this fact. PURPOSE: The purpose was to organizing a nationwide attempt under the Niyantrit Madhumeh Bharat (NMB) programme to break the world record to be the largest Diabetes lesson, to spread awareness among general population. METHODS: Present article represents the perspective of the Chandigarh chapter of NMB programme and its experience in Guinness world record attempt. Diabetes awareness lesson was organized in the city as per the standards defined by the Guinness Book and outcomes of the entire campaign were assessed at the end of the campaign. RESULT: Total 498 individuals participated in the campaign. Among them, 268 participants were questioned at the end of the campaign about the role of Yoga in Diabetes. 247 participants (92%) were agreed that Diabetes can be modified by Yoga and 9 participants (3%) disagreed. The remaining 12 participants (5%) did not give any response. CONCLUSION: We noticed that most of the participants became aware of the role of Yoga in Diabetes.

14.
Ann Neurosci ; 26(2): 75-81, 2019 Apr.
Article in English | MEDLINE | ID: mdl-31975777

ABSTRACT

BACKGROUND: The state of disarray from unhygienic conditions and excessive litter throughout urban highways, alleyways, and byways across rural and urban localities of India is abysmal. Such unsanitary conditions impinge upon the future health and welfare of its citizens, tourists and economic development. PURPOSE: The NRL volunteered PGIMER's campus hygiene initiative" is a pioneering effort spearheaded in compliance with Indian Prime Minister's call that citizens of India work together to establish a cleaner and healthier environment. METHODS: A group of 15 highly motivated students in the Neuroscience Division of the PGIMER, worked together vigorously 2 hours a week to affect a cleaner urban environment in the city. RESULT: The results were national Kayakalp and Skoch award to PGIMER as the cleanest hospital in the country, the vendors or patients no longer litter around the campus, the pot holes have been converted into greener patches, signs board adorn the campus. CONCLUSION: To inspire citizens through faculty- student led sanitation programs.

15.
J Hum Reprod Sci ; 11(3): 291-296, 2018.
Article in English | MEDLINE | ID: mdl-30568361

ABSTRACT

CONTEXT: Physiologically defined homeostatic temperature range for human testis falls within 32°C-37°C and any deviation outside that range can result in impaired steroidogenesis. However, whether temperature deviation within the homeostatic range can affect the steroidogenesis, is still a question of the investigation. AIM: In the present study, we investigated the production of progesterone, testosterone, and vascular endothelial growth factor (VEGF) within the homeostatic temperature range of testis, i.e., 32°C-37°C. SETTING AND DESIGN: We used mouse Leydig tumor cells-1 (MLTC-1) cell line as a model in the present study. MATERIALS AND METHODS: Progesterone and testosterone production by MLTC-1 cells was measured by radioimmunoassay and VEGF production was measured using ELISA. STATISTICAL ANALYSIS USED: Data were analyzed using GraphPad Prism software version 7.04. Student's t-test and ANOVA were used to calculate the P value. P < 0.05 was considered statistically significant. RESULTS AND CONCLUSIONS: Results of our study indicate that there is no significant difference in production of progesterone, testosterone, and VEGF within the homeostatic range of the testicular temperature. Thus, we concluded that testicular cells are able to regulate the steroidogenesis and VEGF production under the homeostatic range of the testicular temperature.

16.
J Complement Integr Med ; 16(2)2018 Nov 07.
Article in English | MEDLINE | ID: mdl-30403654

ABSTRACT

The present study is the case of a breast cancer patient who visited the local unit of the Brahma Kumaris (BK). Conversations with the patient revealed that the patient was being treated in the Post Graduate Institute of Medical Education and Research, Chandigarh, India. The treatment of the patient included the surgery, radiotherapy, and chemotherapy. The patient narrated her ability to manage the psychological stress associated with the cancer treatment. She ascribed it to the positive attitude developed by her practice of the Rajyoga meditation from last 15 years. Rajyoga involved the daily meditation and regular listening to the specified commentary recommended by BK. Patient's health remains stable during the whole treatment process as analyzed by the mammography, histopathology and PET scan. The patient was also able to maintain the total body weight, biochemical and hematological profile within normal physiological range. The present case study is an attempt to provide Rajyoga as supportive means to the maintenance of patient's biochemical and hematological profile, and overall general health during breast cancer treatment.


Subject(s)
Breast Neoplasms/psychology , Breast Neoplasms/therapy , Meditation , Attitude , Breast Neoplasms/blood , Breast Neoplasms/diagnostic imaging , Female , Humans , India , Stress, Psychological
17.
Int J Yoga ; 11(3): 177-185, 2018.
Article in English | MEDLINE | ID: mdl-30233110

ABSTRACT

Despite tremendous advancements in medicine, the number of oral cancer cases continues to increase, and the need for integrating alternate medicine or adopting an integrative approach has become a compelling cost-effective requirement for the management and treatment of diseases. Conventional treatment of oral cancer involves surgery followed by radiotherapy with or without chemotherapy which causes several complications including poor quality of life and high chances of recurrence of cancer. Oral cancer is often linked with obesity which is major risk factors in other cancers. Apart from obesity, oral cancer is thought to have an inverse relation with neurodegenerative disorders presumably because cell death decreases in the former case and increases in the latter. Ancient mind-body techniques such as yoga have not been adequately tested as a tool to synergize the cellular equilibrium pertaining to the treatment of oral cancer. Nerve growth factor (NGF), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) are among the early experimental cellular biomarkers that may be used to probe the modulation of oral cancer, obesity, and neurodegenerative disorders. Yoga has been reported to influence these molecules in healthy individuals but whether their expression can be altered in patients of oral cancer by yoga intervention is the subject of this research being discussed in this review article. Therefore, the present article not only reviews the current status of research studies in oral cancer, obesity, and neurodegenerative disorders but also how these are linked to each other and why the investigations of the putative NGF pathway, involving TNF-α and IL-6, could provide useful clues to understand the molecular effects brought about by yoga intervention in such patients.

18.
J Magn Reson ; 283: 14-21, 2017 10.
Article in English | MEDLINE | ID: mdl-28843057

ABSTRACT

A new solid-state nuclear magnetic resonance (NMR) thermometry sample is proposed. The 207Pb NMR chemical shift of a lead halide perovskite, methylammonium lead chloride (MAPbCl3) is very sensitive to temperature, 0.905±0.010ppmK-1. The response to temperature is linear over a wide temperature range, from its tetragonal to cubic phase transition at 178K to >410K, making it an ideal standard for temperature calibrations in this range. Because the 207Pb NMR lineshape for MAPbCl3 appears symmetric, the sample is ideal for calibration of variable temperature NMR data acquired for spinning or non-spinning samples. A frequency-ratio method is proposed for referencing 207Pb chemical shifts, based on the 1H and 13C frequencies of the methylammonium cation, which are used asan internal standard. Finally, this new NMR thermometer has been used to measure the degree of frictional heating asa function of spinning frequency for a series of MAS rotors ranging in outer diameter from 1.3 to 7.0mm. As expected, the largest diameter rotors are more susceptible to frictional heating, but lower diameter rotors are subjected to higher frictional heating temperatures as they are typically spun at much higher spinning frequencies.

19.
J Cell Biol ; 215(5): 687-703, 2016 Dec 05.
Article in English | MEDLINE | ID: mdl-27903606

ABSTRACT

Although the growth factor progranulin was discovered more than two decades ago, the functional receptor remains elusive. Here, we discovered that EphA2, a member of the large family of Ephrin receptor tyrosine kinases, is a functional signaling receptor for progranulin. Recombinant progranulin bound with high affinity to EphA2 in both solid phase and solution. Interaction of progranulin with EphA2 caused prolonged activation of the receptor, downstream stimulation of mitogen-activated protein kinase and Akt, and promotion of capillary morphogenesis. Furthermore, we found an autoregulatory mechanism of progranulin whereby a feed-forward loop occurred in an EphA2-dependent manner that was independent of the endocytic receptor sortilin. The discovery of a functional signaling receptor for progranulin offers a new avenue for understanding the underlying mode of action of progranulin in cancer progression, tumor angiogenesis, and perhaps neurodegenerative diseases.


Subject(s)
Intercellular Signaling Peptides and Proteins/metabolism , Receptor, EphA2/metabolism , Adaptor Proteins, Vesicular Transport , Capillaries/metabolism , Cell Membrane/metabolism , Enzyme Activation , Genes, Reporter , Green Fluorescent Proteins/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Intercellular Signaling Peptides and Proteins/genetics , Mitogen-Activated Protein Kinases/metabolism , Morphogenesis , Progranulins , Promoter Regions, Genetic/genetics , Protein Binding , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , Solubility
20.
J Biol Chem ; 291(37): 19245-56, 2016 09 09.
Article in English | MEDLINE | ID: mdl-27435676

ABSTRACT

Endorepellin, the C-terminal domain of perlecan, is an angiostatic molecule that acts as a potent inducer of autophagy via its interaction with VEGFR2. In this study, we examined the effect of endorepellin on endothelial cells using atomic force microscopy. Soluble endorepellin caused morphological and biophysical changes such as an increase in cell surface roughness and cell height. Surprisingly, these changes were not accompanied by alterations in the endothelial cell elastic modulus. We discovered that endorepellin-induced autophagic flux led to co-localization of mammalian target of rapamycin with LC3-positive autophagosomes. Endorepellin functioned upstream of AMP-activated kinase α, as compound C, an inhibitor of AMP-activated kinase α, abrogated endorepellin-mediated activation and co-localization of Beclin 1 and LC3, thereby reducing autophagic progression. Functionally, we discovered that both endorepellin and Torin 1, a canonical autophagic inducer, blunted ex vivo angiogenesis. We conclude that autophagy is a novel mechanism by which endorepellin promotes angiostasis independent of nutrient deprivation.


Subject(s)
Autophagy , Heparan Sulfate Proteoglycans/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Peptide Fragments/metabolism , Vascular Endothelial Growth Factor Receptor-2/metabolism , Adenylate Kinase/metabolism , Beclin-1/metabolism , Elastic Modulus , Human Umbilical Vein Endothelial Cells/ultrastructure , Humans , Lactosylceramides/metabolism
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