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1.
J Oral Biol Craniofac Res ; 14(2): 143-151, 2024.
Article in English | MEDLINE | ID: mdl-38333088

ABSTRACT

Objective: To evaluate and compare dentinal tubule penetration and push-out bond strength of BIO-C ION+, AH Plus and NanoSeal-S using Confocal Laser Scanning Microscopy (CLSM) and Universal Testing Machine (UTM). Materials & method: Sixty human mandibular premolars were prepared using ProTaper Gold till F3. Samples were then divided into 3 groups: Group I (n = 20) BIO-C ION + sealer, Group II (n = 20) AH Plus and Group III (n = 20) NanoSeal-S sealer. Groups were then sub-divided into two sub groups: In Subgroup A (n = 10) samples were obturated using single-cone with 0.1 % Rhodamine B dye and in Subgroup B (n = 10) samples were obturated using single cone. The samples were then transversely sectioned into coronal, middle and apical segments, samples in subgroup A & B were then submitted to CLSM analysis and UTM respectively. Results: The Bond Strength data showed following means (MPa): Group I Subgroup B: (BIO-C ION+) coronal (1.64), middle (1.25), apical (0.93); Group II Subgroup B: (AH Plus) coronal (2.20), middle (1.85) apical (1.38) and Group III Subgroup B: (NanoSeal-S) coronal (1.26), middle (0.94), apical (0.58). The dentinal tubule penetration data showed following means: (µm) Group I Subgroup A (BIO-C ION+) coronal (1184.69), middle (997.03), apical (637.26); Group II Subgroup-A AH Plus (864.14) and NanoSeal-S (495.64). Statistical analysis (two-way ANOVA, Tukey's Post Hoc Test) showed significant difference among sealers (p < 0.001) and root canal thirds (p < 0.001). Conclusion: The results of the study concluded that BIO-C ION + sealer showed maximum dentinal tubule penetration and AH Plus demonstrated maximum push-out bond strength.

2.
Clin Gastroenterol Hepatol ; 21(13): 3238-3257, 2023 12.
Article in English | MEDLINE | ID: mdl-37080261

ABSTRACT

BACKGROUND & AIMS: Adenoma detection rate (ADR) is inversely correlated with the risk of interval colon cancer and is a key target for quality improvement in endoscopy units. We conducted a systematic review and meta-analysis to identify and evaluate the effectiveness of interventions that can be implemented at the endoscopy unit level to improve ADRs. METHODS: Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, a systematic search was conducted in MEDLINE, Embase, and Cochrane Central Register of Controlled Trials databases between January 1990 and December 2022 to identify relevant studies. Both randomized controlled trials and observational studies were eligible. Data for the primary outcome of ADR were analyzed and reported on the log-odds scale with 95% CIs using a random-effects meta-analysis model using the empiric Bayes estimator. RESULTS: From 10,778 initial citations, 34 studies were included in the meta-analysis comprising 371,041 procedures and 1501 endoscopists. The provision of report cards (odds ratio [OR], 1.28; 95% CI, 1.13-1.45; P < .001) and the presence of an additional observer to identify polyps (OR, 1.25; 95% CI, 1.09-1.43; P = .002) were associated with significant increases in ADRs whereas multimodal interventions were borderline significant (OR, 1.18; 95% CI, 1.00-1.40; P = .05) and withdrawal time monitoring was not associated significantly with an increase in ADRs (OR, 1.35; 95% CI, 0.93-1.96; P = .11). CONCLUSIONS: The provision of report cards and the presence of an additional observer to identify polyps are associated with improved ADRs and should be considered for implementation in endoscopy facilities.


Subject(s)
Adenoma , Colonic Neoplasms , Polyps , Humans , Colonoscopy/methods , Bayes Theorem , Adenoma/diagnosis , Quality Improvement
3.
J Lasers Med Sci ; 12: e22, 2021.
Article in English | MEDLINE | ID: mdl-34733745

ABSTRACT

Introduction: The conventional chemomechanical procedures are ineffective in complete disinfection of the pulp space due to the complexities of the root canal architecture. The present study aims to compare the efficacy of erbium: YAG laser-activated irrigation, sonic irrigation, and manual dynamic irrigation in the removal of the smear layer through a scanning electron microscope study. Methods: Fifty extracted single rooted mandibular premolars with single canal were used and instrumented until F3 ProTaper rotary file reached the working length. Upon the completion of the canal preparation, each specimen was irrigated with 3 mL of 4% NaOCl for 3 minutes, 3 mL saline for 1 minute and 3 mL of 17% EDTA for 3 minutes. The teeth were assigned to three experimental groups (n=15 each): manual dynamic irrigation, sonic irrigation (EndoActivator), and Er:YAG laser using an X pulse tip. Root canals were sectioned longitudinally and the smear layer at the apical, middle and coronal third was examined under a scanning electron microscope. Smear layer scores were analyzed by Kruskal-Wallis and Mann-Whitney U tests at P = 0.05. Results: The Er:YAG laser group showed significantly lower smear layer scores in the apical third as compared to all other groups. EndoActivator resulted in better cleaning efficacy at the apical area compared to manual dynamic agitation. Conclusion: This study showed results in favor of Er:YAG with an X-pulse tip followed by EndoActivator activation.

4.
J Conserv Dent ; 24(6): 594-598, 2021.
Article in English | MEDLINE | ID: mdl-35558680

ABSTRACT

Background: A myriad of materials and procedures have been recommended for restoring the root-filled teeth with significant loss of tooth structure and the most common method being the use of "posts." Material and Methods: A mesio-occluso-distal preparation was done on eighty intact maxillary first premolars, followed by access preparation and cleaning and shaping with rotary files. All samples were obturated and divided into four groups (n = 20). In Group 1, 3-mm gutta-percha was removed and restored using composite resin; Group 2 - Group 1 + crowns; Group 3 - prefabricated fiber post + crowns; and Group 4 - prefabricated metal post + crowns. All the groups were subdivided into eight groups (n = 10), fractured with 90° vertical and 45° oblique loading by a universal testing machine. Statistical Analysis: Fracture resistance was analyzed and compared by means of two-factorial two-way analysis of variance and Scheffe's post hoc test. Results: Under 90° loading, the fracture load of teeth restored with the Nayyar's composite resin core and crown was greatest. Under 45° loading, the fracture load of teeth restored with prefabricated metallic posts was significantly (P < 0.05) less than that in other groups. Under 45° load, group with fiber post exhibited significantly (P < 0.05) more number of favorable fractures than other groups. Conclusion: Under the conditions of vertical and oblique loadings, fiber post and composite resin core with a full-coverage crown is the most effective restorative modality for protecting the remaining tooth structure in pulpless teeth.

5.
J Pediatr Hematol Oncol ; 42(5): 350-358, 2020 07.
Article in English | MEDLINE | ID: mdl-32576782

ABSTRACT

Acute lymphoblastic leukemia (ALL) is the most common cancer in children and is also seen in adults. Currently, no plasma-based test for the detection of ALL is available. We have cultured the home of a patient with ALL and isolated a mycovirus containing Aspergillus flavus. This culture was subjected to electron microscopy, purification, and mass spectrometry. Using enzyme-linked immunosorbent assay technique, plasma of patients with ALL and long-term survivors of this disease were tested for antibodies, utilizing supernatant of the culture of this organism. The results were compared with 3 groups of controls, including healthy individuals, patients with sickle cell disease, and solid tumors. Using electron microscopy, the isolated A. flavus contained mycovirus particles. In chemical analysis, this organism did not produce any aflatoxin. Using an enzyme-linked immunosorbent assay technique, the supernatant of the culture of the mycovirus containing A. flavus could differentiate ALL patients from each group of controls (P<0.001). These studies provide a new technique for the detection of ALL and may add information for future research regarding leukemogenesis.


Subject(s)
Aspergillosis/complications , Aspergillus flavus/virology , Fungal Viruses/physiology , Plasma/microbiology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Adolescent , Adult , Aspergillosis/microbiology , Aspergillosis/virology , Case-Control Studies , Child , Child, Preschool , Female , Follow-Up Studies , Humans , Male , Precursor Cell Lymphoblastic Leukemia-Lymphoma/blood , Precursor Cell Lymphoblastic Leukemia-Lymphoma/etiology , Prognosis , Young Adult
6.
Int J Clin Pediatr Dent ; 12(3): 205-210, 2019.
Article in English | MEDLINE | ID: mdl-31708616

ABSTRACT

AIM AND OBJECTIVES: This in vitro study evaluated the effect of beveling of enamel on microleakage and shear bond strength of total-etch adhesive system: prime and bond NT and self-etch: adhesive system: XENO V in primary and permanent teeth. MATERIALS AND METHODS: A total of 120 extracted human molars (60 primary and 60 permanent) were selected for the study. For microleakage examination, a sample size of 40 was chosen. Two rectangular slots of equal dimensions were prepared on the buccal surface of each tooth and a bevel was given on either of the slots. Each slot was restored using a composite resin with prior application of the selected bonding agent following which all the samples were soaked in 1% methylene blue dye for 48 hours. Then each tooth was sectioned horizontally and evaluated. The remaining 80 samples were subjected to the shear bond strength test. Class II cavities of standard dimension were prepared and bevel was given on each sample following which selected bonding agent was applied and restored with a composite resin. The specimens were placed in a fixture and the shear bond strength was determined using the universal testing machine. RESULTS: With respect to microleakage, the least was exhibited by beveled preparations in permanent teeth using the self-etch adhesive system and the highest shear bond strength was exhibited by beveled preparations using the total-etch adhesive system in permanent teeth. CONCLUSION: Beveling of enamel improved the marginal integrity and shear bond strength of self-etch and total-etch adhesive systems in both primary and permanent teeth. HOW TO CITE THIS ARTICLE: Patanjali S, Arora A, et al. An In Vitro Study of Effect of Beveling of Enamel on Microleakage and Shear Bond Strength of Adhesive Systems in Primary and Permanent Teeth. Int J Clin Pediatr Dent 2019;12(3):205-210.

7.
BMJ Paediatr Open ; 3(1): e000432, 2019.
Article in English | MEDLINE | ID: mdl-31206078

ABSTRACT

OBJECTIVE: We investigated the content and quality of communication of interservice interprofessional handover between obstetric nurses and neonatal physicians for high-risk deliveries. DESIGN: Observational study. SETTING: Labour and delivery unit at a tertiary care hospital. METHOD: We audio-recorded handovers between obstetric and neonatal teams (n=50) and conducted clinician interviews (n=29). A handover content framework was developed and used to qualitatively code missing core and ancillary content and their potential for adverse events. RESULTS: 26 (52%) handovers missed one or more clinical content elements; a third of the handovers missed at least one core clinical content element. Increase in the number of missed clinical content elements increased the odds of potential adverse events by 2.39 (95% CI1.18 to 5.37). Both residents and nurses perceived handovers to be of low quality and inconsistent and attributed it to the lack of a structured handover process. CONCLUSION: Streamlining handover processes by instituting standardisation approaches for both information organisation and communication can improve the quality of neonatal handovers.

8.
Clin Pediatr (Phila) ; 57(11): 1300-1303, 2018 10.
Article in English | MEDLINE | ID: mdl-29756475

ABSTRACT

BACKGROUND: Documentation and billing/coding are essential to medical practice. Physicians spend significant time documenting to meet coding and medicolegal requirements, potentially reducing time for patient care and learning. We sought to assess time spent charting in pediatric practice and provider understanding and comfort level regarding billing/coding. METHODS: An anonymous web-based survey was emailed to members of American Academy of Pediatrics Section of Pediatric Trainees practicing in the United States. RESULTS: A total of 601 trainees responded to the survey. Thirty-seven percent of trainees spent more than half of patient encounter time documenting in outpatient settings while 62% ( P < .01) in inpatient settings. There was a positive correlation between trainees' apprehension about documentation and reporting increased stress due to documentation ( r = 0.32, P < 0.001). Sixty-two percent respondents had no prior training of billing/coding, and >70% feel necessity of including billing/coding in the medical curriculum ( P < 0.0001). CONCLUSIONS: Our study highlights increasing burden of documentation in practice. Majority of pediatric trainees feel the need to including billing/coding skills as a part of medical curriculum.


Subject(s)
Attitude of Health Personnel , Clinical Coding/statistics & numerical data , Documentation/statistics & numerical data , Internship and Residency , Pediatricians/statistics & numerical data , Practice Patterns, Physicians'/statistics & numerical data , Adult , Female , Humans , Male , Pediatricians/psychology , Stress, Psychological/psychology , Time , United States
9.
Noise Health ; 20(94): 83-89, 2018.
Article in English | MEDLINE | ID: mdl-29785973

ABSTRACT

CONTEXT: The willingness of a person to accept noise while listening to speech can be measured using the acceptable noise level (ANL) test. Individuals with poor ANL are unlikely to become successful hearing aid users. Hence, it is important to enhance the individual's ability to accept noise levels. The current study was an attempt to investigate whether systematic desensitization training can improve the ANL in individuals having high ANL. AIMS: To investigate the effect of systematic desensitization training on ANLs in individuals with normal hearing sensitivity. SETTINGS AND DESIGN: Observational study design. MATERIALS AND METHODS: Thirty-eight normally hearing adults within the age range of 18-25 years participated in the study. Initially, baseline ANL was measured for all participants. Based on the baseline ANL, participants were categorized into three groups; low ANL, mid ANL, and high ANL. The participants with high ANL were trained using systematic desensitization procedure whereas, individuals with low and mid ANL did not undergo any training and served as the comparison groups. After the training period, ANL was measured again for all the participants. STATISTICAL ANALYSIS USED: Repeated measures of analysis of variance with follow up paired "t" test. RESULTS: Analysis revealed a significant main effect of systematic desensitization training on ANL. There was a significant improvement in ANL in participants with high ANL. However, there was no significant difference in ANL between baseline and follow-up session in individuals with low and mid ANL. CONCLUSIONS: Systematic desensitization training can facilitate ANL, thereby enhancing the individual's ability to accept the noise levels. This enhanced ANL can facilitate better hearing aid fitting and acceptance.


Subject(s)
Desensitization, Psychologic , Loudness Perception , Noise , Speech Perception , Acoustic Stimulation , Adolescent , Adult , Hearing , Hearing Aids , Humans , Patient Acceptance of Health Care , Psychoacoustics , Young Adult
10.
Int J Clin Pediatr Dent ; 11(6): 536-538, 2018.
Article in English | MEDLINE | ID: mdl-31303744

ABSTRACT

Usually first mandibular molars have one mesial and distal root but in some cases there are anatomical variations wherein the number of roots and root canals vary. Presence of an additional lingual root distally in mandibular molars is called radix entomolaris (RE). Appropriate diagnosis is must before starting with root canal procedure in these teeth to ensure successful treatment outcome. The report describes the endodontic management of mandibular molar with RE. HOW TO CITE THIS ARTICLE: Arora A, Arya A, Chauhan L, Thapak G. Radix Entomolaris: Case Report with Clinical Implication. Int J Clin Pediatr Dent, 2018;11(6):536-538.

11.
Noise Health ; 19(90): 222-226, 2017.
Article in English | MEDLINE | ID: mdl-28937016

ABSTRACT

BACKGROUND: Intact hearing is essential for medical students and physicians for communicating with patients and appreciating internal sounds with a stethoscope. With the increased use of (PMSs), they are exposed to high sound levels and are at a risk of developing hearing loss. The effect of long term personal music system (PMS) usage on auditory sensitivity has been well established. Our study has reported the immediate and short term effect of PMS usage on hearing especially among medical professionals. OBJECTIVE: To assess the effect of short term PMS usage on distortion product otoacoustic emissions (DPOAE) among medical professionals. MATERIALS AND METHOD: 34 medical students within the age range of 17-22 years who were regular users of PMS participated in the study. All participants had hearing thresholds <15 dBHL at audiometric octave frequencies. Baseline DPOAEs were measured in all participants after 18 h of non-usage of PMS. One week later DPOAEs were again measured after two hours of continuous listening to PMS. DPOAEs were measured within the frequency range of 2 to 12 kHz with a resolution of 12 points per octave. Output sound pressure level of the PMS of each participant was measured in HA-1 coupler and it was converted to free field SPL using the transformations of RECD and REUG. RESULTS: Paired sample t test was used to investigate the main effect of short term music listening on DPOAE amplitudes. Analysis revealed no significant main effect of music listening on DPOAE amplitudes at the octave frequencies between 2 to 4 KHz (t67 = -1.02, P = 0.31) and 4 to 8 KHz (t67 = 0.24, P = 0.81). However, there was a small but statistically significant reduction in DPOAE amplitude (t67 = 2.10, P = 0.04) in the frequency range of 9 to 12 kHz following short term usage of PMS. The mean output sound pressure level of the PMS was 98.29. CONCLUSION: Short term exposure to music affects the DPOAE amplitude at high frequencies and this serves as an early indicator for noise induced hearing loss (NIHL). Analysis of output sound pressure level suggests that the PMSs of the participants have the capability to induce hearing loss if the individual listened to it at the maximum volume setting. Hence, the medical professionals need to be cautious while using PMS.


Subject(s)
Hearing Loss, Noise-Induced/etiology , Music/psychology , Noise/adverse effects , Otoacoustic Emissions, Spontaneous/physiology , Students, Medical/psychology , Adolescent , Audiometry, Pure-Tone , Auditory Threshold/physiology , Female , Humans , Male , Risk Factors , Time Factors , Young Adult
12.
Pediatr Cardiol ; 37(6): 1137-40, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27160097

ABSTRACT

This study aimed to understand the knowledge, attitudes and confidence level related to critical congenital heart disease (CCHD) screening among pediatric residents. Pediatric residents were assessed via an anonymous survey related to CCHD guidelines and procedures as set out by the New York State Department of Health. The survey was emailed to pediatric residents at a large academic institution. A teaching intervention was performed after the initial survey, which was followed by an identical after-intervention survey. Forty-two residents responded to the pre-intervention survey (n = 42), and forty post-intervention (n = 40). The mean composite knowledge score was 76 % pre-intervention and 92 % post-intervention, p < .001. Pre-intervention only 12 % of the respondents could answer all the questions correctly which increased to 60 % post-intervention. Confidence among residents regarding guidelines increased from 38 to 95 % post-intervention, p < .001. There was a positive correlation between the residents who felt confident of the guidelines and who answered correctly, r = .514, p < .001, n = 82. There was no significant difference between knowledge, attitudes or confidence level scores by year of training or the gender of the residents. Our study demonstrated a significant gap of knowledge among residents related to CCHD screening mandated by New York State health law. There was also a corresponding lack of confidence demonstrated by the residents in the guidelines. These data suggest that residents would benefit from further education on the proper implementation of a CCHD screening program. Further multicenter studies are warranted to assess similar gaps in other residency training programs in New York and wherever these screenings guidelines have been adopted.


Subject(s)
Heart Diseases , Child , Health Knowledge, Attitudes, Practice , Humans , Internship and Residency , New York , Surveys and Questionnaires
13.
J Biosci ; 40(2): 313-24, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25963259

ABSTRACT

Epithelial mesenchymal transition (EMT) of lens epithelial cells (LECs) may contribute to the development of posterior capsular opacification (PCO), which leads to visual impairment. Andrographolide has been shown to have therapeutic potential against various cancers. However, its effect on human LECs is still unknown. The purpose of this study is to evaluate the effect of andrographolide on EMT induced by growth factors in the fetal human lens epithelial cell line (FHL 124). Initially the LECs were treated with growth factors (TGF-beta 2 and bFGF) to induce EMT. Subsequently these EMT-induced cells were treated with andrographolide at 100 and 500 nM concentrations for 24 h. Our results showed that FHL 124 cells treated with growth factors had a significant decrease in protein and m-RNA levels of epithelial markers pax6 and E-Cadherin. After administering andrographolide, these levels significantly increased. It was noticed that EMT markers alpha-SMA, fibronectin and collagen IV significantly decreased after treatment with andrographolide when compared to the other group. Treatment with andrographolide significantly inhibited phosphorylation of ERK and JNK. Cell cycle analysis showed that andrographolide did not arrest cells at G0/G1 or G2/M at tested concentrations. Our findings suggest that andrographolide helps sustain epithelial characteristics by modulating EMT markers and inhibiting the mitogen-activated protein kinase (MAPK) signalling pathway in LECs. Hence it can prove to be useful in curbing EMT-mediated PCO.


Subject(s)
Cataract/prevention & control , Diterpenes/pharmacology , Epithelial-Mesenchymal Transition/drug effects , Lens, Crystalline/metabolism , MAP Kinase Signaling System/drug effects , Actins/metabolism , Cell Line , Collagen Type IV/metabolism , Epithelial Cells/metabolism , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Fibronectins/metabolism , Flavonoids/pharmacology , Humans , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , Phosphorylation/drug effects
14.
J Conserv Dent ; 17(1): 35-9, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24554858

ABSTRACT

AIM: The aim of this study was to compare the canal transportation, canal centering ability, and time taken for preparation of curved root canals after instrumentation with ProFile GT Series X (GTX) files, Revo-S files, twisted files, and Mtwo files by using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: Forty mesiobuccal canals of mandibular molars with an angle of curvature ranging from 20 to 40 were divided according to the instrument used in canal preparation into four groups of 10 samples each: GTX (group I), Revo-S (group II), twisted file (group III), and Mtwo (group IV). The teeth were instrumented according to manufacturer's guidelines, with all groups being prepared to size 30, 0.06 taper master apical file. Canals were scanned using an i-CAT CBCT scanner (Imaging Science International, Hatfield, PA, USA) before and after preparation to evaluate the transportation and centering ratio at 3 mm, 5 mm, 7 mm, 9 mm, and 11 mm from the apex. The data collected were evaluated using one-way analysis of variance (ANOVA) with Tukey's honestly significant difference (HSD) test. RESULTS: Twisted file system showed significantly least canal transportation and highest canal centering ability values as compared to GTX, Revo-S, and Mtwo file systems. Overall, GTX, Revo-S, and Mtwo showed comparable results with respect to canal transportation and centering ability. CONCLUSION: The innovative method of manufacturing the TF system resulted in superior shaping ability in curved canals, with the instruments remaining more centered and producing less canal transportation than GTX, Revo-S, and Mtwo file systems.

15.
Invest Ophthalmol Vis Sci ; 54(9): 6224-33, 2013 Sep 17.
Article in English | MEDLINE | ID: mdl-23970468

ABSTRACT

PURPOSE: To explore different molecular factors impairing the activities of superoxide dismutase (SOD) isoforms in senile cataractous lenses. METHODS: Enzyme activity of SOD isoforms, levels of their corresponding cofactors copper (Cu), manganese (Mn), zinc (Zn), and expression of mRNA transcripts and proteins were determined in the lenses of human subjects with and without cataract. DNA from lens epithelium (LE) and peripheral blood was isolated. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) followed by sequencing was carried out to screen somatic mutations. The impact of intronic insertion/deletion (INDEL) variations on the splicing process and on the resultant transcript was evaluated. Genotyping of IVS4+42delG polymorphism of SOD1 gene was done by PCR-restriction fragment length polymorphism (RFLP). RESULTS: A significant decrease in Cu/Zn- and Mn-SOD activity (P < 0.001) and in Cu/Zn-SOD transcript (P < 0.001) and its protein (P < 0.05) were found in cataractous lenses. No significant change in the level of copper (P = 0.36) and an increase in the level of manganese (P = 0.01) and zinc (P = 0.02) were observed in cataractous lenses. A significant positive correlation between the level of Cu/Zn-SOD activity and the levels of Cu (P = 0.003) and Zn (P = 0.005) was found in the cataractous lenses. DNA sequencing revealed three intronic INDEL variations in exon4 of SOD1 gene. Splice-junction analysis showed the potential of IVS4+42delG in creating a new cryptic acceptor site. If it is involved in alternate splicing, it could result in generation of SOD1 mRNA transcripts lacking exon4 region. Transcript analysis revealed the presence of complete SOD1 mRNA transcripts. Genotyping revealed the presence of IVS4+42delG polymorphism in all subjects. CONCLUSIONS: The decrease in the activity of SOD1 isoform in cataractous lenses was associated with the decreased level of mRNA transcripts and their protein expression and was not associated with either modulation in the level of enzyme cofactors or with INDEL variations.


Subject(s)
Cataract/enzymology , Coenzymes/metabolism , Superoxide Dismutase/metabolism , Aged , Blotting, Western , Cataract/genetics , Copper/metabolism , DNA Mutational Analysis , Epithelium, Corneal/enzymology , Female , Gene Expression Regulation, Enzymologic , Genotype , Humans , INDEL Mutation , Male , Manganese/metabolism , Middle Aged , Polymorphism, Single Nucleotide , Protein Isoforms/metabolism , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Superoxide Dismutase-1 , Zinc/metabolism
16.
Indian J Med Res ; 137(1): 117-24, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23481060

ABSTRACT

BACKGROUND & OBJECTIVES: Cytoskeletal proteins are deregulated during oxidative stress and cataract formation. However, estrogen which protects against cataract formation and harmful effects of oxidative stress has not been tested on the cytoskeleton of lens epithelial cells (LECs). The current study was undertaken to assess if the protection rendered to LECs by estrogen was mediated by preserving the cytoskeletal proteins. METHODS: Oxidative stress was induced by 50 µM of H 2 O 2 in cultured goat LECs (gLECs) and effect of 1 µM 17ß-estradiol (E 2 ) was tested. After treatment, morphological analysis of cells was carried out using haematoxylin-eosin staining and cell density was also quantified. Cell viability was determined using Hoechst (Ho), YO-Pro (YP) and propidium iodide (PI). F-actin and vimentin were localized using phalloidin and anti-vimentin antibody, respectively, and viewed under fluorescence microscopy. Vimentin was further analysed at protein level by Western blotting. RESULTS: H 2 O 2 led to increased condensation of nucleus, cell death and apoptosis but these were prevented with pre- and co-treatment of E 2 with increase in cell viability (P<0.001). E 2 also prevented H 2 O 2 mediated depolymerization of cytoskeleton but was not able to reverse the changes when given after induction of oxidative stress. INTERPRETATION & CONCLUSIONS: Our findings showed that E 2 helped in preventing deteriorating effect of H 2 O 2 , inhibited cell death, apoptosis and depolymerisation of cytoskeletal proteins in LECs. However, the exact mechanism by which estrogen renders this protection to cytoskeleton of lens epithelial cells remains to be determined.


Subject(s)
Cataract/pathology , Epithelial Cells/drug effects , Lens, Crystalline/drug effects , Oxidative Stress , Animals , Apoptosis/drug effects , Cataract/etiology , Cataract/metabolism , Cell Survival/drug effects , Cells, Cultured , Cytoskeleton/drug effects , Cytoskeleton/pathology , Epithelial Cells/cytology , Estradiol/administration & dosage , Estrogens/administration & dosage , Goats , Humans , Hydrogen Peroxide/toxicity , Lens, Crystalline/cytology , Membrane Potential, Mitochondrial/drug effects , Reactive Oxygen Species/metabolism
17.
J Cataract Refract Surg ; 39(4): 617-23, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23522583

ABSTRACT

PURPOSE: To compare changes in the incision's histomorphology and denaturation of collagen I in rabbit eyes having microcoaxial phacoemulsification through 2.2 mm and 1.8 mm incision-compatible systems. DESIGN: Randomized experimental trial. SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. METHODS: Thirty rabbit eyes were randomized into Group 1 (microcoaxial phacoemulsification through 2.2 mm incisions using Infiniti system [torsional ultrasound]) and Group 2 (microcoaxial phacoemulsification through 1.8 mm incisions using Stellaris system [longitudinal ultrasound]). Each group was then divided into 3 subgroups of 5 eyes each based on 1 of the 3 intervention options: phacoemulsification only, intraocular lens (IOL) insertion only, and phacoemulsification with IOL insertion. Left eyes were randomized for microcoaxial phacoemulsification, and right eyes were treated as controls. RESULTS: After phacoemulsification, eyes in Group 1 showed loss of epithelium at the roof of the incisions and Descemet membrane detachment at the floor of the incisions. These findings did not change after IOL insertion. After phacoemulsification, eyes in Group 2 showed loss of epithelium, but Descemet membrane remained attached. There was a longitudinal split in the incision's stroma in the direction of internal entry. The stromal damage increased after IOL implantation. Immunofluorescence studies showed no obvious irregularities in the arrangement of collagen I in either group. A dot blot analysis showed significant denaturation of collagen I in Group 2. CONCLUSION: The histomorphology of the 2.2 mm system incision showed localized Descemet membrane detachment and endothelial cell loss. The 1.8 mm system incision showed exaggerated stromal damage after IOL insertion.


Subject(s)
Cornea/pathology , Cornea/surgery , Lens Implantation, Intraocular , Microsurgery/methods , Phacoemulsification/methods , Animals , Collagen Type I/metabolism , Cornea/metabolism , Fluorescent Antibody Technique, Indirect , Microscopy, Fluorescence , Rabbits , Wound Healing
18.
J Biosci ; 37(6): 979-87, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23151788

ABSTRACT

Specimens of the anterior lens capsule with an attached monolayer of lens epithelial cells (LECs) were obtained from patients (n=52) undergoing cataract surgery. Specimens were divided into three groups based on the type of cataract: nuclear cataract, cortical cataract and posterior subcapsular cataract (PSC). Clear lenses (n=11) obtained from donor eyes were used as controls. Expression was studied by immunofluorescence, real-time PCR and Western blot. Statistical analysis was done using the student's t-test. Immunofluorescence results showed punctate localization of Cx43 at the cell boundaries in controls, nuclear cataract and PSC groups. In the cortical cataract group, cytoplasmic pools of Cx43 without any localization at the cell boundaries were observed. Real-time PCR results showed significant up-regulation of Cx43 in nuclear and cortical cataract groups. Western blot results revealed significant increase in protein levels of Cx43 and significant decrease of ZO-1 in all three cataract groups. Protein levels of alpha-catenin were decreased significantly in nuclear and cortical cataract group. There was no significant change in expression of beta-catenin in the cataractous groups. Our findings suggest that ZO-1 and alpha-catenin are important for gap junctions containing Cx43 in the LECs. Alterations in cell junction proteins may play a role during formation of different types of cataract.


Subject(s)
Cataract/metabolism , Connexin 43/metabolism , Lens, Crystalline/metabolism , Zonula Occludens-1 Protein/metabolism , alpha Catenin/metabolism , beta Catenin/metabolism , Base Sequence , Blotting, Western , Case-Control Studies , DNA Primers , Epithelial Cells/metabolism , Fluorescent Antibody Technique , Humans , Real-Time Polymerase Chain Reaction
19.
J Cataract Refract Surg ; 38(5): 833-9, 2012 May.
Article in English | MEDLINE | ID: mdl-22520307

ABSTRACT

PURPOSE: To compare incision integrity after clear corneal microcoaxial phacoemulsification using longitudinal and torsional ultrasound (US). SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. DESIGN: Prospective randomized experimental clinical trial. METHODS: Part 1 comprised an experimental study of rabbit eyes. Group 1 received longitudinal US. Group 2 received torsional US. The right eye of each rabbit served as a control. Samples were processed for histomorphology and collagen I denaturation by immunofluorescence. Part 2 comprised a clinical trial of patients. Group 1 received torsional US. Group 2 received longitudinal US. At the end of surgery, trypan blue 0.0125% was instilled. After 2 minutes, 0.1 mL of aqueous was aspirated and its optical density measured. RESULTS: In part 1, incision histomorphology was comparable in both modalities. Collagen denaturation tests (immunofluorescence, dot blot analysis) showed no irregularity in collagen arrangement in either group. In Group 2, Descemet membrane was detached and endothelial cells were minimal at the roof of the incision. In part 2, trypan blue ingress into the anterior chamber was significantly greater in Group 1 than in Group 2 (mean 3.40 + 0.6 log units versus and 3.77 + 0.82 log units) (P<.007). CONCLUSIONS: Incision histomorphology in the torsional group showed minimal Descemet membrane detachment and minimal endothelial cell loss at the roof of the incision. Minimal ingress of trypan blue into the anterior chamber was observed with torsional US, indicating better wound integrity than with longitudinal US. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.


Subject(s)
Cornea/diagnostic imaging , Cornea/surgery , Phacoemulsification/methods , Surgical Flaps/pathology , Animals , Aqueous Humor/metabolism , Collagen Type I/metabolism , Coloring Agents/pharmacokinetics , Corneal Endothelial Cell Loss/diagnosis , Descemet Membrane/pathology , Double-Blind Method , Female , Fluorescent Antibody Technique, Indirect , Humans , Lens Implantation, Intraocular , Male , Middle Aged , Prospective Studies , Rabbits , Trypan Blue/pharmacokinetics , Ultrasonography , Wound Healing
20.
Mol Vis ; 18: 64-73, 2012.
Article in English | MEDLINE | ID: mdl-22259225

ABSTRACT

PURPOSE: To evaluate the level of matrix metalloproteinase (MMP)-2 and MMP-9 activities in patients with steroid induced posterior subcapsular cataract (PSC). METHODS: This prospective, observational study comprised of 156 patients having either steroid induced PSC (n=50) or non-steroidal PSC (n=106) were performed to evaluate the level of MMP-2 and MMP-9 activities in the lens epithelial cells (LECs) and the serum. Anterior lens capsules harboring LECs were obtained during phacoemulsification and peripheral blood was collected from patients before administration of anesthesia. Serum was separated by centrifugation at 10,000× g for 15 min at 4 °C. The LECs and serum samples were processed to analyze MMP-2 and MMP-9 activities using succinylated gelatin assay. Quantitative real time-PCR (qRT-PCR) was performed to determine the mRNA levels of MMP-2 and MMP-9 in LECs. The mRNA levels were expressed as a ratio, using the delta-delta method for comparing the relative expression results between cases with steroid induced PSC and cases with non-steroidal PSC. MMP-2 and MMP-9 levels were also compared in the two groups using immunolocalization. RESULTS: The level of MMP-2 and MMP-9 activity was found to be high in LECs and serum of cases with steroid induced PSC. Further in all steroid induced cases, a 1.4 fold increase was observed in MMP-2 activity in LECs and a 1.4 fold increase in MMP-9 activity in the serum. Both qRT-PCR and immunolocalization showed increased expression of MMP-2 and MMP-9 activity. CONCLUSIONS: MMP-2 and MMP-9 activity in both LECs and serum was significantly higher in cases with steroid induced PSC. The possible use of MMP-9 as a non-invasive biomarker in ascertaining the presence of steroid induced PSC should be evaluated using a larger sample size.


Subject(s)
Capsule Opacification/blood , Epithelial Cells/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Posterior Capsule of the Lens/drug effects , Adolescent , Adult , Aged , Biomarkers/blood , Capsule Opacification/chemically induced , Child , Cyclosporine/adverse effects , Dexamethasone/adverse effects , Epithelial Cells/pathology , Female , Gene Expression , Glucocorticoids/adverse effects , Humans , Immunosuppressive Agents/adverse effects , Male , Matrix Metalloproteinase 2/blood , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 9/blood , Matrix Metalloproteinase 9/genetics , Middle Aged , Posterior Capsule of the Lens/pathology , Prednisolone/adverse effects , Prospective Studies , RNA, Messenger/blood
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