85 research outputs found
Rapid Non-genomic Actions of Cortisol in Fish
No full textCortisol, the primary glucocorticoid (GC) in teleosts signals through either the genomic pathway, by activating the intracellular glucocorticoid receptor (GR) and/or the mineralocorticoid receptor (MR), or through non-genomic pathways. However, the mechanism of action of non-genomic cortisol signalling is far from clear, and there is a complete lack of consensus as to the physiological significance of this rapid action. The major goal of this thesis was to determine the mode of action of cortisol in bringing about rapid changes in intracellular Ca2+ levels [iCa2+] as a non-genomic response, and to assess the physiological consequences of this rapid effect at the cellular and organismal level. This was tested using rainbow trout (Oncorhynchus mykiss) hepatocytes (non-excitable) and zebrafish (Danio rerio) muscle (excitable) explant as in vitro and ex vivo models, respectively. The results indicate that a direct interaction of cortisol with the calcium release-activated (CRAC) channel may be responsible for the rapid increase in [iCa2+] due to cortisol. This increase in [iCa2+] with cortisol played a key role in the translocation of intracellular GR to the plasma membrane. This was evident because inhibition of the CRAC channel abolished the colocalization of GR to the caveolin-1 on the plasma membrane of hepatocytes. Cortisol also stimulated a rapid increase in [iCa2+] in zebrafish muscle explants, underscoring a conserved non-genomic role for cortisol in rapidly increasing [iCa2+] in excitable and non-excitable cells. A zebrafish tail-fin amputation model was used to investigate the physiological role of non-genomic and genomic cortisol signalling in regulating epimorphic regeneration. Cortisol rapidly increased [iCa2+] at the amputated site, and this corresponded with an increased ORAI1 (protein subunit of CRAC) expression at the site. Also, an increase in cell proliferation of ORAI1 expressing cells and blastema formation was evident at 24 h in response to cortisol stimulation, but absent in the GR knockout and MR knockout larvae, suggesting that this CRAC channel protein may be both non-genomically and genomically regulated by cortisol. Taken together, this thesis represents a significant contribution to the mechanism of action of rapid non-genomic effects of cortisol, which is essential to our understanding of the role of cortisol in stress adaptation in vertebrates
Quorum Sensing Signals Produced by Heterotrophic Bacteria in Black Band Disease (BBD) of Corals and Their Potential Role in BBD Pathogenesis
Get PDFBlack band disease (BBD) of corals is a temperature dependent, highly virulent, polymicrobial disease affecting reef-building corals globally. The microbial consortium of BBD is primarily comprised of functional physiological groups that include photosynthetic cyanobacteria, sulfate reducers, sulfide oxidizers and a vast repertoire of heterotrophic bacteria. Quorum sensing (QS), the cell-density dependent communication phenomenon in bacteria, is known to induce expression of genes for a variety of virulence factors in diseases worldwide. Microbes capable of QS release signals such as acyl homoserine lactones (AHLs) and autoinducer-2 (AI-2), which coordinate microbial interaction. The focus of the present study was to investigate the presence and potential role of QS in BBD pathogenicity, utilizing culture dependent and independent methodologies. Isolates across coral health states including BBD, were screened for production of QS signals, and AHL and AI-2 production capabilities were analyzed via LC-MS/MS. The effect of temperature on AHLs was also examined. Additionally, antimicrobial production capabilities of isolates were tested. BBD metagenomes were utilized to screen for sequences related to QS, antimicrobial synthesis, and antimicrobial resistance genes. BBD isolates represented a significantly higher proportion of isolates capable of producing QS signals in comparison to healthy coral isolates. Several AHLs produced by coral derived bacterial cultures were identified, and three AHLs, specifically 3OHC4, 3OHC5 and 3OHC6, showed a significant increase in production at an elevated temperature of 30 °C, which correlates with increased BBD incidence on reefs with increasing water temperature. Most of the BBD cultured isolates were identified as vibrios. Several sequences related to QS, antimicrobial synthesis and resistance genes were detected in the BBD metagenomes. Based on the findings of this study, a model for potential microbial interactions amongst BBD heterotrophs, centered around QS, is proposed. Taken together, the findings from this study provide a clearer understanding of the potential role of QS in BBD, and serve as the basis for further studies aimed at elucidating the pathogenesis of an intricate coral disease
Fragment tailoring strategy to design novel chemical entities as potential binders of novel corona virus main protease
No full textThe
recent pandemic of novel corona virus (nCoV) infections (COVID19) has put the
world on serious alert. The main protease of nCov (nCov-MP) cleaves the long
polyprotein chains to release functional proteins required for replication of
the virus and thus is a potential drug target to design new chemical entities
in order to inhibit the viral replication in human cells. The current study
employs state of art computational methods to design novel molecules by linking
molecular fragments which specifically bind to different constituent sub-pockets
of the nCov-MP binding site. A huge library of 191678 fragments was screened
against the binding cavity of nCov-MP and high affinity fragments binding to
adjacent sub-pockets were tailored to generate new molecules. These newly
formed molecules were further subjected to molecular docking, ADMET property
filters and MM-GBSA binding free energy calculations to select 17 best
molecules (named as MP-In1 to Mp-In17), which showed comparable binding
affinities and interactions with the key binding site residues as the reference
ligand. The complexes of these 17 molecules and the reference molecule with
nCov-MP, were subjected to molecular dynamics simulations, which assessed the
stabilities of their binding with nCov-MP. Fifteen molecules were found to form
stable complexes with nCov-MP. These novel chemical entities designed
specifically according to the pharmacophoric requirements of nCov-MP binding
pockets showed good synthetic feasibility and returned no exact match when
searched against chemical databases. Considering their interactions, binding
efficiencies and novel chemotypes, they can be further evaluated as potential
starting points for nCov drug discovery.
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Samples.docx
No full text
An observational cross-sectional analytical study was carried out in the Department of Prosthodontics, Crown and Bridge in Sharad Pawar Dental College & Hospital, Wardha, Maharashtra, India to evaluate and correlate shades of Indian Human Skin with a custom made silicone shade guide for Indian population to be used in rehabilitation of maxillofacial defects.
Room temperature vulcanising medical grade silicone, Technovent (Figure No.:1) was selected as the base material for the shade guide and facial prosthesis owing to its superiority over conventional acrylic resin and polyvinyl chloride.
The colours of human skin are merely due to presence of three coloring pigments present namely, carotene, Haemoglobin and Melanin. The Spectrophotometric studies has enabled us to divide the shade of human skin into three types with the Hues corresponding to those of the above three pigments Carotene (C), Haemoglobin (H) and Melanin (M). For the purpose of fabrication of a shade tab the following Core Hues were related and subdivided according to their decreasing amount of Values.
Once the shade tabs were made the skin on the colour of the patients with defect was correlated with the tab. The patients were instructed about the study and were consented to particicpate in the same.
Fabrication of shade tabs -
A total of 21 shade tabs each with measurements of 10 mm x 3 mm were fabricated using the stainless steel mould (Figure No.2) which was specially fabricated for this purpose. The silicon elastomer was mixed in a conventional way and a Digital Balance was used to weigh the pigments. Each drop of Pigment which was dispensed weighed around 0.03 gm. Corresponding Intrinsic Silicon Pigments (Figure No.:3) were used to obtain the corresponding shade tabs.
Further tabs were created by mixing the Core Pigments. Carotene- Melanin ( CM ), Melanin-Haemoglobin ( MH ), Haemoglobin-Carotene ( HC ) and Haemoglobin-Melanin-Carotene ( MHC ) which is the maximum combination possible (Figure no. 4). Further shade tabs were created by decreasing the amounts of the pigments by 50% consistently from the previous shade and they were arranged in the descending order and were numbered from 1 to 3 respectively. The pigmented silicon was packed and poured in the mould spaces which was closed with a lid and tightened with screws and cured for period of 30 mins. The pigmented silicon was retrieved from the moulds and finished with the help of a fine grit stone.
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Figure No.:1 – Technovent Silicone Figure No.:2 – Intrinsic stains
elastomer
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Figure .: 3 – Stainless Steel Mould
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Figure No.:4 – Seven Core Pigments
Sample size and selection-
A cross-sectional study was conducted on 100 random Indians aged between 19-25 years of age for skin color matching using the skin tab inclusive of male and female participants in the ratio of 1:1. A Convenience sampling was done to include the volunteering patients in the study. Accordingly the patients were informed about the evaluation and consent was obtained. Other criteria on which the patient was dependent was that the patient should be devoid of any type of skin disease, post surgical scars, any deformity of the facial skin , history of burn injury of face or history of bleaching recently. The patients which were selected for the study were also advised to maintain abstinence of the use of skin products 2 hours prior to the evaluation. Patients with known systemic disorders which can cause Hypopigmentation or Hyperpigmentation or patients which complain of recent skin tanning or patients with a history of radiation therapy were excluded. The patients who were not willing to participate in the study were excluded.
Evaluation of color matching-
The fabricated skin shade tab was selected according to the skin shade of the patient and was kept in close approximation to the skin of the patient. Care was taken to evaluate the shade of the skin of the patient in natural daylight conditions. Evaluation was done independently by two other experienced Dental Professionals. The evaluators for the study underwent thorough check up for colour deficiency. The selected evaluators underwent the “Ishihra colour test (Pseudo Isochromatic Plate (PIP) Colour Vision Test)” and were reported with physiologic vision. Scores for the individual group were noted by all the evaluators on the customised sheet. The depiction of colour for patients was done three times to avoid errors. The colour which was matched at the very first sight was identified at that very instant. Time was not Prolonged for colour matching as it might fatigue the eyes.
The colour matching was assessed visually on the basis of a score sheet with the use of a scoring system. The interpretation of the scoring record was as follows:
1. Total mismatch and unacceptable.
2. Almost same and acceptable.
3. Adequate colour match and satisfactory.
4. Accurate colour match.
STATISTICAL ANALYSIS
Descriptive statistics were done (Table No. 1). The data is represented in number and percentages.
A total of 100 subjects fulfilling the inclusion criteria were included in the study. There were no cases (0.0%) with total mismatch and unacceptable. Seventy (70.0%) cases had accurate colour match, 27 (27.0%) had adequate colour match and was satisfactory and only 3 (3.0%) had almost same and were acceptable. Software used was Microsoft Excel for Mac OS High Sierra 10.13.
Table 1: Matching grades of custom-made silicone shade guide to Indian human skin shades
Grades
N
%
Total mismatch and unacceptable
0
0.0
Almost same and acceptable
3
3.0
Adequate colour match and satisfactory
27
27.0
Accurate colour match
70
70.0
Total
100
100.0
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Table 1: Matching grades of custom-made silicone shade guide to Indian human skin shades
RESULTS
The statistics revealed that the color matching for 70% of the analysed cases showed accurate match. The statistical analysis has made the revelation that the shade guide can be used to effect for shade matching with intrinsic stains for fabrication of Maxillofacial Prosthesis. About 3% of the patients showed almost same and acceptable shade match. About 27% of the patients showed adequate colour match and satisfaction. No patient showed total mismatch and unacceptable skin shade selection. </p
Environmental Conditions and Test Procedures for Airborne Equipment - A Comprehensive guide to Aircraft Systems Designers
No full textEnvironmental conditions and test procedures for airborne13; equipment is a standard document from -Radio Technical Commission for13; Aeronauticsquot;, describing the various test procedures of avionic systems13; of civilian aircraft. A detailed study and the classification of these13; test procedures is a must for selection of avionic instruments. In an13; aircraft there are equipments on and inside the aircraft at different environmental conditions. As per this the equipments are to be selected13; for different categories suitable to the requirements and tested for13; quot;determining compliance with equipment performance standardquot;.13; This document describes Radio Technical Commission for Aeronautics13; 160-B and 160-C (RTCA\160-B and RTCA\160-C) in a comprehensive manner13; for environmental conditions and test procedures in general and for13; Light Aircrafts in particular. There are 23 tests to be conducted and13; categories are to be selected for each equipment depending upon the13; position in which the the equipment is located in or on the aircraft.13; From the position the environmental conditions can be determined and a13; category selection can be made. This type of selection will help in13; reducing the Certification procedures
Impact of Medicare Accountable Care Organizations (ACOs) and Physician Social Networks on Urologic Cancer Surgery Outcomes
No full textThesis (Ph.D.)--University of Rochester. School of Medicine & Dentistry. Dept. of Public Health Sciences, 2022.In 2012, CMS introduced the voluntary Medicare Shared Savings Program (MSSP) in the Accountable Care Organizations (ACO) model to control cost and improve quality and outcomes for Medicare beneficiaries. Despite increase in urologic cancer cases and surgical care accounting for 30% of total U.S. health care costs, the role of Medicare ACOs and physician social networks has been unexplored in urologic cancer care. Previous literature has only focused on participation in the Medicare Shared Savings Program ACOs and quality of care outcomes. Evaluating ACO’s focus on contracts, rules and regulations coupled with the impact of physician interactions (through shared patients) can help better understand post discharge outcomes and care transition for urologic cancer surgery patients. Using quasi-experimental design and network analysis methods, the dissertation first assessed the impact of Medicare ACO participation on post-acute care (PAC) use and spending, and post-surgical outcomes for urologic cancer surgeries, a) Prostate Cancer b) Bladder Cancer and c) Kidney Cancer. Second, this dissertation investigated the impact of MSSP participation and physician patient-sharing networks on post-surgical readmissions and complications in Medicare patients undergoing prostatectomy, cystectomy, and nephrectomy. Third, this dissertation evaluated associations between Medicare ACOs and physician patient-sharing networks on post-acute care (PAC) use and spending in Medicare beneficiaries undergoing urologic cancer surgeries. Transaction Cost Economics, Social Network Theory and AHRQ framework for organizational response to incentives served as the basis for theoretical and conceptual framework. For bladder cancer surgery (cystectomy), Medicare ACO participation was associated with significantly reduced overall post-acute care use, but not with changes in readmission or complication rates. For prostate cancer (prostatectomy) and kidney cancer surgery (nephrectomy), we found no significant association between hospital participation in Medicare ACOs and PAC use or post-surgical outcomes. In Medicare ACOs, higher disassortativity and higher internal patient sharing were significantly associated with lower readmissions while higher physician degree centrality was associated with significantly higher complications following prostatectomy, cystectomy, and nephrectomy. Although MSSP participation was independently associated with significantly lower 90-day readmissions and 90-day complications following cystectomy and 30-day readmissions following prostatectomy and nephrectomy, higher disassortativity reinforced these associations. Similar results were obtained for disassortativity and PAC use for all three procedures. This is the first study to demonstrate that both physician patient-sharing relationships and formal participation in the MSSP together could help identify strategies for ACOs to improve post-discharge care coordination and reduce fragmented care. It is hoped that results of this study will influence ACO participation in MSSP and role of physician networks in improving post-surgical outcomes and post-acute care use and spending in urologic cancer. Furthermore, the study results might have implications by encouraging more specialists to participate, thereby improving value for urologic cancer care
Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori.
No full textGalactosidases, ubiquitous in nature, are complex carbohydrate-active enzymes and find extensive applications in food, pharma, and biotechnology industries. The present study deals with the production of galactosidases from fungi by solid-state fermentation. Fifteen fungi were screened and Aspergillus awamori
(MTCC 548), exhibited the highest α and β-galactosidase activities of 75.11±0.29 U/g and 155.34±
1.26 U/g, respectively. 30 g of wheat bran substituted with 6% defatted soy flour, at 28°C, pH 5.0 for
120 h, was established as the optimum production conditions by one-factor approach. The enzyme was purified to homogeneity with an apparent mass of 118 ± 2 kDa by ammonium sulfate precipitation (50–80%),
ion exchange and hydrophobic interaction chromatography. Specific activities for α and β-galactosidase
were 22 and 74 U/mg, respectively. Optimum temperature and pH ranges for enzyme activities were
55–60 °C, 5.0–5.5, respectively. The thermal inactivation mid-point was 65 °C. The purified enzyme not
only exhibited α and β-galactosidase activities, but also exhibited β-xylosidase and β-glucosidase activities,
indicating the enzyme has broad substrate specificity. Sequence analysis by in-gel digestion and tandem
mass spectrometry (MS/MS) revealed that the enzyme was a probable β-galactosidase A, belonging to glycoside hydrolase 35 family, and is being reported for the first time
A new thermostable rhizopuspepsin: Purification and biochemical characterisation
No full textThe new thermostable fungal aspartic protease was produced through solid-state fermentation from Rhizopus azygosporus
(MTCC 10195). The protease was purified by a three-tandem steps of ammonium sulfate precipitation (25−65% saturation), ion exchange (DEAE sepharose CL 6B) chromatography, and gel filtration (Sephacryl S 200) chromatography. The optimum pH and temperature for protease activity were 4 and 52 ± 1.8
◦ C, respectively. The enzyme was unusually thermostable, as it retained 50 % of its activity beyond 85 ◦ C after 20
min of incubation. The apparent molecular weight was 33.2 ± 2.3 kDa with a final specific activity of 86.6 U/mg.
The enzyme was rich in β sheets (63 %) and was inhibited by pepstatin A, indicating that it is an aspartic protease. MS/MS analysis revealed the enzyme is an endopeptidase cleaving the C-terminus of Phe, Leu, Lys, Val, His, Glu, Met, and Tyr. Amino-terminal sequencing, coupled with in-gel trypsin digestion and MS analysis revealed that the enzyme was being reported for the first time with “experimental evidence at protein-level”
Comparison of Carotid Ultrasound Parameters in Patients with Rheumatoid Arthritis and Control Subjects: A Cross-sectional Study
No full textIntroduction: Rheumatoid Arthritis (RA) is considered one of the risk factors for Cardiovascular Disease (CVD). Carotid ultrasound is used to assess the extent of subclinical atherosclerosis in patients with RA. Very few studies have evaluated all carotid ultrasound findings, such as Carotid Intima-media Thickness (CIMT), carotid diameters like Luminal Diameter (LD) and Interadventitial Diameter (IAD), and plaque characteristics in patients with RA.
Aim: To assess and compare the various carotid parameters in RA patients with controls and also to evaluate their association with age and duration of onset of the disease.
Materials and Methods: A single-centre cross-sectional study was conducted at the Department of Radiology, GCS Medical College and Hospital, Ahmedabad, Gujarat, India, involving 40 patients with RA (Group-RA) and 40 controls (Group-C). Patients were further divided into three age subgroups. Carotid ultrasound parameters like CIMT, LD, IAD, and plaque were evaluated in both groups. Continuous variables were analysed using the t-test, and categorical data were analysed using the Chi-squared test. A p-value of < 0.05 was considered statistically significant.
Results: Age- and sex-matched controls were included with 28 females and 12 males in both the study groups. The mean LD and IAD were significantly higher in the RA group than in controls (Mean LD- 5.88±0.97 mm vs 5.26±1.08 mm with p=0.009; Mean IAD- 6.85±0.89 mm vs 6.30±0.87 mm with p=0.006). The mean CIMT was higher in the RA group but not statistically significant (0.57±0.13 mm vs 0.54±0.12 mm with p=0.256). Further subgroup analysis showed that all carotid parameters were statistically significant in the 31-60 years age group as compared to 18-30 years and 61-80 years subgroups. The effect of the duration of the disease (>5 years) on the carotid parameters was also significantly higher in the 30-60 age group (p<0.001).
Conclusion: The present study showed that various carotid ultrasound parameters can be used as a screening tool in the follow-up of RA patients to detect early changes in atherosclerosis, with LD and IAD having superior predictive capability. The study further showed that carotid ultrasound parameters have a better predictive value in the 31-60 years age group
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