Comparative transcriptomic insights into molecular mechanisms of the susceptibility wheat variety MX169 response to Puccinia striiformis f. sp. tritici (Pst) infection.

Stripe rust of wheat is caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst). Breeding durably resistant wheat varieties by disrupting the susceptibility (S) gene has an important impact on the control of wheat stripe rust. Mingxian169 (MX169) showed strong stripe rust susceptibility to all the races of Pst. However, molecular mechanisms and responsive genes underlying susceptibility of the wheat variety MX169 to Pst have not been elucidated. Here, we utilized next-generation sequencing technology to analyze transcriptomics data of "MX169" and high-resistance wheat "Zhong4" at 24, 48, and 120 h post-inoculation (hpi) with Pst. Comparative transcriptome analysis revealed 3,494, 2,831, and 2,700 differentially expressed genes (DEGs) at different time points. We observed an upregulation of DEGs involved in photosynthesis, flavonoid biosynthesis, pyruvate metabolism, thiamine metabolism, and other biological processes, suggesting their involvement in MX169's response to Pst. DEGs encoding transcription factors were also identified. Our study suggested the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst. IMPORTANCE: Our study suggests the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst.

Comparative Evaluation of Antimicrobial Efficacy of Diode Laser (Continuous Mode), Diode Laser (Pulse Mode), and 5.25% of Sodium Hypochlorite in Disinfection of Root Canal: A Short Study.

Introduction: The success of endodontic treatment is achieved by the complete elimination of pulpal infection and protection from future invasion of microorganisms. Due to the complex structure of the root canal, complete eradication of microorganisms is not possible and is a major challenge for successful endodontic treatment. Therefore, microbiological studies are needed to explore the effect of various disinfection methods. Aim and objective: The aim of this study is to compare the effectiveness of root canal disinfection by using a diode laser (in pulse and continuous modes) and sodium hypochlorite by microbiological assessment. Materials and methods: Forty-five patients were selected and randomly divided into three groups. After gaining patency to the root canal the first sample from the root canal was taken using a sterile absorbent paper point and transferred to a sterile tube containing a normal saline medium. Biomechanical preparation was performed with Dentsply Protaper hand files of each corresponding group and was disinfected with group I (diode light amplification by stimulated emission of radiation (LASER) 980 nm with 3 W in continuous mode for 20 seconds), group II (diode LASER 980 nm with 3 W in pulse mode for 20 seconds), group III (irrigated with 5.25% of sodium hypochlorite for 5 minutes). Pre- and post-samples of each group were inoculated on sheep blood agar and examined for any bacterial growth. After the microbial evaluation of the total microbial count of pre- and post-samples, the data obtained were tabulated and statistically analyzed. Results: The data were evaluated and analyzed using analysis of variance (ANOVA) on Statistical Package for the Social Sciences (SPSS) software. Groups I, II, and III all three groups showed significant differences (p < 0.01) and an overall reduction in the microbial count at postbiomechanical preparation (BMP) as compared to pre-BMP with the highest being in laser in continuous mode (group I) (91.9%), followed by 5.25% sodium hypochlorite (group III) (86.5%) and LASER in pulse mode (group II) (72.0%) the least. Conclusion: The study concluded that the diode laser in continuous mode is more efficacious than the diode laser in pulse mode and 5.2% sodium hypochlorite, respectively. How to cite this article: Mishra A, Koul M, Abdullah A, et al. Comparative Evaluation of Antimicrobial Efficacy of Diode Laser (Continuous Mode), Diode Laser (Pulse Mode), and 5.25% of Sodium Hypochlorite in Disinfection of Root Canal: A Short Study. Int J Clin Pediatr Dent 2022;15(5):579-583.

Effect of Addition of Incipient Carious Lesions in Caries Diagnostic Criteria on Prevalence of Caries in North Indian Preschool Children of East Lucknow Region.

AIM AND OBJECTIVE: To investigate the effect of the addition of incipient carious lesions into the WHO caries index on the prevalence of caries in preschoolers of East Lucknow city. STUDY DESIGN: A cross-sectional observational study was conducted on 530 participants in the age-group of 3-6 years. MATERIALS AND METHODS: The participants were randomly selected from nursery schools of the East Lucknow region. The dental caries status was assessed according to WHO and Ekstrand's criteria. In addition, the prevalence of cavitated and incipient carious lesions was also evaluated. The data were then subjected to statistical analysis. STATISTICS: Chi-square test and Wilcoxon signed-rank test were performed to carry out statistical analysis. p value < 0.005 was taken as statistically significant. RESULTS: Out of 530 participants examined, caries prevalence by WHO and Ekstrand's criteria was (36.8%) and (57.4%), respectively. The mean dmft values according to WHO criteria and Ekstrand's criteria were 1.16 and 1.60, respectively. The prevalence of cavitated carious lesions was higher (66.27%) when compared with that of incipient carious lesions (33.73%). CONCLUSION: A significant number of incipient carious lesions observed in the present study is an indication to include these lesions in caries diagnostic criteria. This would help in determining the exact prevalence of caries and aid in implementing proper preventive protocols for remineralization of incipient carious lesions. HOW TO CITE THIS ARTICLE: Asad F, Singh N, Rathore M, et al. Effect of Addition of Incipient Carious Lesions in Caries Diagnostic Criteria on Prevalence of Caries in North Indian Preschool Children of East Lucknow Region. Int J Clin Pediatr Dent 2021;14(5):644-647.

Alternaria alternata causing Alternaria Leaf Spot of Cucumis melo (Muskmelon) in Pakistan.

In July 2019, leaf spot symptoms were observed on muskmelon (Cucumis melo L.) cv. Jackball-1 plants in an experimental field of 2.02 ha with a disease incidence of 30% (31°26'05.4"N 73°04'30.3"E) at the University of Agriculture, Faisalabad, Pakistan. Early symptoms consisted of small, circular, brown, necrotic spots 1 to 2 mm in size covering 10 to 30% of the leaf blade, which gradually enlarged and developed concentric rings. To identify the causal agent of the disease, a total of 20 symptomatic leaves were collected. Small pieces removed from the margin between healthy and diseased tissues were surface disinfected in 70% ethanol for 2 min, rinsed three times with sterile distilled water, plated on Potato dextrose agar and incubated at 25 ± 2°C with a 12-h photoperiod. Morphological observations were made on 7-day-old single-spore cultures. The colonies initially appeared white and then turned olive-green. All 20 fungal isolates were characterized by small, short-beaked, multicellular conidia. The conidia were ellipsoidal or ovoid and measured 11.5 to 30 µm × 7.5 to 15 µm (n = 50) with longitudinal and transverse septa. Conidia were produced on short conidiophores in chains. The beaks were short (often less than one-third the body length) and conical or cylindrical. These morphological features concur with the description of Alternaria alternata (Fr.) Keissler (Woudenberg et al. 2013). For molecular identification, genomic DNA of four representative isolates (HMSMZA 07, 08, 09, 10) were extracted and PCR amplification of the internal transcribed spacer (ITS)-rDNA, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and translation elongation factor-1 alpha (TEF-1α) gene regions were performed (White et al. 1990, Berbee et al. 1999, Carbone & Kohn, 1999) respectively. The obtained sequences were deposited in GenBank with accession numbers MT253643.1-MT253646.1 (ITS-rDNA), MT318260.1-MT318263.1 (GAPDH), and MT318280.1-MT318283.1 (TEF-1α). BLASTn analysis of HMSMZA 07 sequences showed 100% identity with ITS rDNA (MN615420.1), GAPDH (MK637438.1) and TEF-1α (MN807795.1) sequences of A. alternata. To confirm pathogenicity, 5-6 weeks-old Muskmelon (Cucumis melo L.) cv. Jackball-1 plants (true leaf stage) were sprayed until runoff (1.5 to 2 ml per plant) with A. alternata conidial suspension (106 conidia/ml; obtained from 1 week-old cultures) amended with 0.1% (vol/vol) of Tween 20 using an atomizer in the green house. The experiment included four A. alternata isolates inoculated onto three muskmelon plants per each isolate, whereas control plants (n = 3) were sprayed with sterile distilled water amended with 0.1% Tween 20. The plants were incubated at 25 ± 2°C in a greenhouse and the experiment was conducted twice. After 5 to 7 days post inoculation, necrotic leaf spots were observed on the inoculated plants and A. alternata was reisolated and confirmed by morphological and molecular (ITS) features. No disease was observed on control plants. Previously, A. alternata on muskmelon has been reported in Pakistan (Ahmad et al. 1997), however this study provides a detailed description of disease symptoms, morphological and molecular identity of the causal agent including completion of Koch's postulates. The disease could represent a threat for muskmelon crop in Pakistan due to its increasing cultivation and therefore warrants the need to develop disease management strategies.

Occurrence of Leaf Spot Caused by Alternaria alternata on Eggplant (Solanum melongena) in Pakistan.

Eggplant (Solanum melongena L.) is a popular vegetable that is grown in both tropical and subtropical regions all year long. The crop is cultivated on small family farms and is a good source of income for resource-limited farmers in Pakistan. In early May 2019, leaf spots on eggplant (cv. Bemisaal) were observed in an experimental field (31°26'14.0"N 73°04'23.4"E) at the University of Agriculture, Faisalabad, Pakistan. Early symptoms were small, circular, brown, necrotic spots uniformly distributed on leaves. The spots gradually enlarged and coalesced into large, nearly circular or irregularly shaped spots that could be up to 3 cm in length. The center of the spots was light tan, surrounded by a dark brown ring, a chlorotic halo, and tended to split in the later developmental stages. Disease incidence was approximately 35% in the infected field. The causal agent of this disease was isolated consistently by plating surface sterilized (1% NaOCl) sections of symptomatic leaf tissue onto potato dextrose agar (PDA). After 6 days incubation at 25°C with a 12-h photoperiod, fungal colonies had round margins and the cottony mycelia were dark olivaceous with a mean diameter of 7.5 cm. For conidial production, the fungus was grown on potato carrot agar (PCA) and V8 agar media under a 16-h/8-h light/ dark photoperiod at 25°C. Conidiophores were septate, light to olive golden brown with a conidial scar, from which conidia were produced. Conidia were borne singly or in short chains and were obpyriform to obclavate, measured 29 ± 4.8 × 13.25 ± 2.78 µm (n=30) with zero to three longitudinal and two to six transversal septa. The morphological characters matched those of Alternaria alternata (Fr.) Keisel (Simmons et al. 2007). DNA was extracted using the DNAzol reagent (Thermo Fisher Scientific MA, USA). For molecular identification, internal transcribed spacer (ITS) region between ITS1 and ITS2, actin gene (ß-Actin), translation elongation factor (TEF-1α) gene, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene of two representative isolates (JLUAF1 and JLUAF2) were amplified with primers ITS1/ITS4 (White et al. 1990), ß-Actin 512 F/783 R, EF1-728F/-986R (Carbone et al. 1999), and gpd1/gpd2 (Berbee et al. 1999), respectively. The sequences were deposited in GenBank (accession nos. MT228734.1 and MT228735.1 for ITS; MT260151.1 and MT260152.1 for ß-Actin, MT260163.1 and MT260164.1, for TEF-1a, and MT260157.1 and MT260158.1 for GAPDH). BLASTn analysis of these sequences showed 100% identity with the sequences of A. alternata for ITS rDNA, ß-Actin, TEF-1α, and GAPDH, respectively. Based on the morphological characters and DNA sequences, the leaf spot isolates of eggplant were identified as A. alternata. To confirm the pathogenicity on eggplant, six-week old healthy potted eggplants of cv. Bemisaal were sprayed at the true leaf stage with conidial suspensions of A. alternata (106 conidia/ml; obtained from 1-week-old cultures) amended with 0.1% (vol/vol) of Tween 20 until runoff (1.5 to 2 ml per plant) using an atomizer in the greenhouse. Three plants were inoculated with each of the two isolates (JLUAF1 and JLUAF2), whereas three control plants were sprayed with sterile distilled water amended with 0.1% Tween 20. The plants were incubated at 25 ± 2°C in a greenhouse, and the experiment was conducted twice. After 10 days of inoculation, each isolate induced leaf spots which were similar to typical spots observed in the field, whereas the control plants remained symptomless. The fungus was re-isolated from symptomatic tissues. Re-isolated fungal cultures were morphologically and molecularly identical to A. alternata, thus fulfilling Koch's postulates. Previously, A. alternata has been reported to cause leaf spots on eggplant in India (Raina et al. 2018). To our knowledge, this is the first report of A. alternata causing leaf spot on eggplant in Pakistan. The disease could represent a threat for eggplant crops due to its increasing cultivation. It is important to develop disease management strategies for Alternaria alternata causing leaf spot of Eggplant in Pakistan.

First Report of Aspergillus niger causing Black rot of Grapes in Pakistan.

In June 2015 & 2016, a postharvest survey of table grapes (Vitis vinifera) cv. King's Ruby, was carried out in five different commercial fruit markets of Rawalpindi (33°38'19.2″N, 73°01'45.0″E) district, Punjab Province. Symptoms appeared as brownish lesions with black sporulation on grapes berries. The incidence of these symptoms on bunches ranged from 12 to 17% at all sites. Symptomatic tissue pieces were surface-sterilized with 0.1% sodium hypochlorite (NaOCl) for 30 seconds, rinsed three times with sterile distilled water, dried on filter paper for 45 seconds, and incubated on potato dextrose agar (PDA) at 25°C. After 3 days, dark brown to black mycelium were formed on PDA media. A total of 24 isolates were examined morphologically. The apex of the conidiophore was observed to be radiate. Vesicles were found to be spherical and covered with irregular metulae and phialides. Conidia were globose or subglobose measured (3.14 µm ± 2.24 in averaged diameter: n=50), dark brown to black, with roughened cell walls. The conidiophores were also smooth-walled, hyaline, and became melanized toward the vesicle. These characteristics of the fungus were similar to those described for Aspergillus niger van Tiegh (de Hoog et al. 2000). For molecular identification, the internal transcribed spacer (ITS) region, beta-tubulin (Bt) gene and partial RNA polymerase II largest subunit (RPB2) gene of representative isolate (Asp.n02) was amplified using primers ITS1/ITS4, BT2a/BT2b and RPB2-6F/RPB2-7R respectively (White et al., 1990; Glass & Donaldson, 1995; Liu et al. 1999). Sequences were deposited in GenBank (ITS, MN658871; Bt2, MT117924; and RPB2, MT318289). Based on BLAST analysis, sequences of the ITS region, Bt2 genes, and RPB2 gene showed 99 to 100% similarity of isolate Asp.n02 to Aspergillus niger (Accession Nos. MK307680.1, MN195121.1, MF078661.1 for ITS gene, MN567299.1, MK451029.1, MK451020.1 for Bt2 gene, and MK450788.1, MK450790.1 for RPB2 gene). To complete Koch's postulates, 10-µl aliquots of spore suspensions (106 spores/ml) of isolate: Asp.n 02 was pipetted onto three non-wounded and four wounded (5 mm diam) asymptomatic grape berries cv. King's Ruby (seven berries per isolate), Sterile distilled water was applied to asymptomatic berries similaries to serve as a negative control (Ghuffar et al. 2018; Jayawardena et al. 2018). Berries were incubated at 25 ± 2°C in sterile moisture chambers, and the experiment was conducted twice. Brownish lesions leading to black sporulation similar to the original symptoms were observed on both wounded and non-wounded inoculated berries after 3 days, whereas no symptoms were recorded on the negative control. The morphology of the fungus that was re-isolated from each of the inoculated berries was identical to that of the original cultures. Aspergillus niger was reported previously in Europe and Israel causing mycotoxin (Ochratoxin A) OTA production on Table grapes (Bau et al. 2006). To our knowledge, this is the first report of Aspergillus niger causing black rot of grapes in Pakistan. This finding will help to plan effective disease management strategies against the black rot of grapes in Pakistan.

A Comparative Evaluation of Shear Bond Strength of Seventh- and Eighth-Generation Self-etch Dentin Bonding Agents in Primary Teeth: An In Vitro Study.

AIM: To compare the shear bond strength of seventh- and eight-generation self-etch dentin bonding agents in primary teeth using universal testing machine. MATERIALS AND METHODS: Forty extracted sound human primary molars were collected and randomly divided into two groups of 20 sample each. Shear bond strength of seventh-generation bonding agent [ADPER Single Bond Universal, (group II)] and eighth-generation bonding agent [Futurabond DC, VOCO Germany, (group I)] were calculated by using Universal Testing Machine (PTC/O83/ME, INSTRON, USA) and expressed in megapascals (MPa). Recorded data were compiled and subjected to statistical analysis using Student's t test. RESULTS: The mean shear bond strength of group I (eighth-generation dentin bonding agent) and group II (seventh-generation dentin bonding agent) ranged from 22.10 to 37.10 MPa and 19.80 to 30.30 MPa, respectively Student's t test showed significantly different and higher (8.7%) shear bond strength in group I when compared to group II (p value = 0.017). CONCLUSION: Shear bond strength of eighth-generation dentin bonding agent (Futurabond DC) was better than seventh-generation dentin bonding agent (Adper Single Bond Universal). CLINICAL SIGNIFICANCE: An effective bond to tooth would reduce marginal microleakage, bacterial penetration, postoperative sensitivity, possibility of pulpal inflammation, and preserve tooth structure by allowing minimal cavity preparation. HOW TO CITE THIS ARTICLE: Mishra A, Koul M, Upadhyay VK, et al. A Comparative Evaluation of Shear Bond Strength of Seventh- and Eighth-generation Self-etch Dentin Bonding Agents in Primary Teeth: An In Vitro Study. Int J Clin Pediatr Dent 2020;13(3):225-229.

Oral mucocele in infant with an unusual presentation.

Mucocele is a benign lesion characterised by an extravasation or retention of mucous secretion in submucosal tissue from minor salivary glands. The occurrence of mucocele is common in the oral cavity, with more than 70% of cases on lower labial mucosa. The occurrence of mucocele on the tongue is considered rare and accounted for only 2.25% in the previous literature. Histopathological examination plays a great role in the confirmation of the clinical diagnosis. Management of mucocele is at times challenging. This paper highlights a rare case of mucocele in a 5-month-old infant who was successfully managed under day-care anaesthesia.

Effect of head posture on tooth contacts in dentate and complete denture wearers using computerized occlusal analysis system.

PURPOSE: The purpose of the study was to assess and compare the occlusal contacts in dentate and edentulous patients wearing complete denture with varying head posture. MATERIALS AND METHODS: Ad hoc sampling of 30 subjects (15 dentate and 15 edentulous) based on inclusion and exclusion criteria was done. Subjects were divided into two groups: dentate and edentulous. Each group was further divided into two subgroups based on two head postures-upright 90° and ventroflexed 30°. For recording of every posture, a new sensor was used, and the subject was asked to clench on the sensor in maximum intercuspation position at the two head postures. RESULTS: Data were summarized as mean ± standard error and compared by Student's t-test using SPSS software (windows version 17.0 IBM corporation, New York, USA). A statistically significant correlation between head posture and contact area was found in dentate and denture wearers, i.e., tooth contact area varies with head posture. CONCLUSION: It was concluded that the occlusal contacts vary at different head posture in dentate as well as in denture wearers. With ventroflexion, the number of tooth contact decreased as compared to upright-erect position in both groups. Clinical implication - since the number of tooth contacts varies with varying head postures, it is recommended that the balancing of the contacts should be done at varying head postures.

A rare case of three single rooted permanent second molars: A diagnosis with cone beam computed tomography (CBCT).

The aim of this article is to present a case with an unusual morphology of maxillary second molars on both sides and a mandibular second molar on right side in the same patient which was observed during routine endodontic procedure. Three second permanent molars had single root, single canal and an accurate assessment of this unusual morphology was made with the help of radiographs and cone beam computed tomography (CBCT). This report extends the range of known possible anatomical variations to include teeth with lesser number of roots and canals.

Comparative Evaluation of Electronic Apex Locators and Radiovisiography for Working Length Determination in Primary Teeth in vivo.

AIMS: The aim of this study was to evaluate the effectiveness of two different varieties of electronic apex locators and radiovisiography (RVG) for working length determination in primary teeth. MATERIALS AND METHODS: A total of 30 primary teeth indicated for pulpectomy in children aged 3 to 8 years were randomly selected and subjected to working length determination using two varieties of electronic apex locators and RVG separately. The data were then subjected to statistical analysis. RESULTS: A very strong correlation between electronic measurement methods and RVG length was observed. CONCLUSION: Radiovisiography and apex locators are equally effective in determining working length in primary teeth. How to cite this article: Abdullah A, Singh N, Rathore MS, Tandon S, Rajkumar B. Comparative Evaluation of Electronic Apex Locators and Radiovisiography for Working Length Determination in Primary Teeth in vivo. Int J Clin Pediatr Dent 2016;9(2):118-123.