Cold-Active Enzymes and Their Potential Industrial Applications-A Review.

More than 70% of our planet is covered by extremely cold environments, nourishing a broad diversity of microbial life. Temperature is the most significant parameter that plays a key role in the distribution of microorganisms on our planet. Psychrophilic microorganisms are the most prominent inhabitants of the cold ecosystems, and they possess potential cold-active enzymes with diverse uses in the research and commercial sectors. Psychrophiles are modified to nurture, replicate, and retain their active metabolic activities in low temperatures. Their enzymes possess characteristics of maximal activity at low to adequate temperatures; this feature makes them more appealing and attractive in biotechnology. The high enzymatic activity of psychrozymes at low temperatures implies an important feature for energy saving. These enzymes have proven more advantageous than their mesophilic and thermophilic counterparts. Therefore, it is very important to explore the efficiency and utility of different psychrozymes in food processing, pharmaceuticals, brewing, bioremediation, and molecular biology. In this review, we focused on the properties of cold-active enzymes and their diverse uses in different industries and research areas. This review will provide insight into the areas and characteristics to be improved in cold-active enzymes so that potential and desired enzymes can be made available for commercial purposes.

Prevalence of aspergillus skin hypersensitivity in allergic rhinitis.

Allergic rhinitis (AR) is a chronic allergen specific, IgE-mediated hypersensitivity disorder which significantly impairs the quality of life in affected patients. Many aeroallergens and molds are responsible for AR. This study was  conducted to find prevalence of Aspergillus fumigatus skin hypersensitivity in patients of AR by skin prick test (SPT).  150 clinically diagnosed AR patients visiting our OPD were enrolled. Skin hypersensitivity for Aspergillus f. was done by SPT in all the enrolled patients. Chi square test and Student’s t-test were applied for statistical analysis. Out of 150 patients, 60 (40%) were positive for Aspergillus fumigatus SPT. Majority of the positive patients had persistent AR and among those 39/60 (65%) had moderate to severe persistent AR and 12/60 (20%) had mild persistent AR.  Our study concluded that there is high (40%) prevalence of Aspergillus f. skin hypersensitivity among AR patients. The hypersensitivity to Aspergillus f. was found more in severe AR patients and it was statistically significant.

Application of CBNAAT (Xpert MTB/RIF assay) in new smear negative pulmonary tuberculosis patients.

Presumptive pulmonary tuberculosis (PTB) patients whose sputum are detected to be smear negative for acid fast bacilli (AFB) present a significant challenge for a treating physician. Initiating these patients on anti tuberculous treatment (ATT) on empirical basis is not a good strategy as many were found to be sputum culture for tuberculosis negative on further evaluation according to many previous studies. In India due to resource limited settings and lack of knowledge about newest diagnostic modalities patients are often initiated only on the basis of characteristic clinical symptoms and chest radiographic abnormalities. This study was conducted to identify the advantage of application of sputum cartridge based nucleic acid amplification test (CBNAAT) in sputum AFB smear negative presumptive pulmonary TB patients. Our study concluded that clinical symptoms and radiological characteristics cannot differentiate TB patients from non-TB patients. Treating patients only on empirical basis would have resulted in unnecessary treatment of 41 patients.

Development, optimization and evaluation of curcumin loaded biodegradable crosslinked gelatin film for the effective treatment of periodontitis.

OBJECTIVE: Aim of the present study was to prepare curcumin (CUR) loaded biodegradable crosslinked gelatin (GE) film to alleviate the existing shortcomings in the treatment of periodontitis. SIGNIFICANCE: Gelatin film was optimized to provide anticipated mucoadhesive strength, mechanical properties, folding endurance, and prolonged drug release over treatment duration, for successful application in the periodontitis. METHODS: The film was developed by using solvent casting technique and "Design of Experiments" approach was employed for evaluating the influence of independent variables on dependent response variables. Solid-state characterization of the film was performed by FTIR, XRD, and SEM. Further, prepared formulations were evaluated for drug content uniformity, surface pH, folding endurance, swelling index, mechanical strength, mucoadhesive strength, in vitro biodegradation, and in vitro drug release behavior. RESULTS: Solid state characterization of the formulation showed that CUR is physico-chemically compatible with other excipients and CUR was entrapped in an amorphous form inside the smooth and uniform film. The optimized film showed degree of crosslinking 51.04 ± 2.4, swelling index 138.10 ± 1.25, and folding endurance 270 ± 3 with surface pH around 7.0. Crosslinker concentrations positively affected swelling index and biodegradation of film due to altered matrix density of the polymer. Results of in vitro drug release demonstrated the capability of the developed film for efficiently delivering CUR in a sustained manner up to 7 days. CONCLUSIONS: The developed optimized film could be considered as a promising delivery strategy to administer medicament locally into the periodontal pockets for the safe and efficient management of periodontitis.

Screening of ionically crosslinked chitosan-tripolyphosphate microspheres using Plackett-Burman factorial design for the treatment of intrapocket infections.

OBJECTIVE: Application of Plackett-Burman factorial design to investigate the effect of processing factors in the fabrication of ionically crosslinked chitosan-tripolyphosphate (CS-TPP) microspheres. SIGNIFICANCE: Microspheres were screened and optimized to provide maximum process yield (PY), encapsulation efficiency (EE), and time for 80% drug release (T80%) and minimum burst and particles size (PS), for successful application in periodontitis. METHODS: Processing factors viz. method of preparation (MOP), CS, TPP, crosslinking time (CT), agitation (AS), and drying technique (DT) were selected. Solid state characterization was performed by Fourier-Transform infrared (FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Mucoadhesion, cytocompatibility, and stability of microspheres were also evaluated. RESULTS: Pareto analysis and analysis of variance, screened most significantly (p < .05) impacting process factors on selected responses. The optimized microspheres demonstrated: o/w emulsification method, CS (2.5%), TPP (5%), CT (120 min), AS (2000 rpm), and DT (freeze-dried), and provided PY- 95.67%, PS- 168.45%, EEOZ- 85.56%, EEDX- 91.34%, BOZ- 15.26%, BDX- 12.91%, TOZ- 47.09 and TDX- 67.95 minutes. FTIR illustrated compatibility between excipients and complexation of CS and TPP. XRD and DSC showed loss of crystallinity of entrapped drugs in microspheres. Biphasic drug release was observed for four days with non-Fickian kinetics. Furthermore, microspheres exhibited good mucoadhesivity (82.51%), antimicrobial activity against Staphylococcus aureus and Escherichia coli, cytocompatibility for L929 cells, and long-term stability. CONCLUSIONS: Therefore, CS-TPP microspheres were found mucoadhesive, safe, stable and provided controlled and prolonged release of drugs. These properties confirmed its high potential and applicability in chronic periodontitis.

Development and Evaluation of Biodegradable Chitosan Films of Metronidazole and Levofloxacin for the Management of Periodontitis.

Metronidazole (MZ) and levofloxacin (LF) are widely employed for treatment of periodontitis, but high oral dose and resistance development after long-term oral administration limit their use. The aim of this study was to alleviate shortcomings in the treatment of periodontitis by fabrication of intrapocket, biodegradable films of chitosan (CS) loaded with MZ and LF meant for inserting into periodontal pockets to treat infections. The films were developed by solvent casting technique using propylene glycol as plasticizer and glutaraldehyde as crosslinking agent. Their physical characteristics, such as drug content, surface pH, swelling index, and folding endurance, exhibited results within limit. Further, FTIR and DSC studies revealed stability of films and compatibility between drugs and excipients. SEM images of films showed the presence of free drug particles on the surface causing burst effect. In vitro release in McIlvaine buffer pH 6.6 was of sustained nature assisted by the burst effect. CS and crosslinking agent concentrations negatively affected drug release and positively affected T90 (time for releasing 90% of the drug) due to altered matrix density. In contrast, the plasticizer concentration increases membrane permeability and hence increased drug release, lowering T90. Crosslinked films demonstrated sustained release up to 7 days. The antibacterial efficacy of films was tested on Staphylococcus aureus and Escherichia coli, indicating good antibacterial activity. Clinical trials on patients proved the therapeutic efficacy of the films by a significant (p < 0.05) decrease in the clinical markers of periodontitis, i.e. gingival index, plaque index and pocket depth. Conclusively, the films of MZ and LF were successful tools for the management of periodontitis.

Global outbreaks of foodborne hepatitis A: Systematic review and meta-analysis.

Hepatitis A Virus (HAV) is a significant threat in terms of food safety. A systematic literature search with the research question "What are the clinical outcomes of foodborne Hepatitis A virus infections?" was conducted. The pooled estimate of the outcomes-mortality, hospitalization, and severity rates, along with a 95% confidence interval (CI), was estimated. After screening, 33 studies were included for the data extraction and meta-analysis. The pooled prevalence of hospitalization among the HAV-positive patients was estimated to be 32% (95% CI 21-44), with high heterogeneity (I2 = 98%, p < 0.01). Australia had the highest hospitalization rate, with 82%, followed by Europe (42%). The hospitalization rate showed a significantly increasing trend (beta = 0.015, p=0.002) over the period. The pooled prevalence of mortality among the HAV-positive patients was estimated to be <1%, with low heterogeneity (I2 = 5%, p = 0.39). A wide range of food products were linked with the HAV outbreaks.

Transfer learning for image classification using VGG19: Caltech-101 image data set.

Image classification is getting more attention in the area of computer vision. During the past few years, a lot of research has been done on image classification using classical machine learning and deep learning techniques. Presently, deep learning-based techniques have given stupendous results. The performance of a classification system depends on the quality of features extracted from an image. The better is the quality of extracted features, the more the accuracy will be. Although, numerous deep learning-based methods have shown enormous performance in image classification, still due to various challenges deep learning methods are not able to extract all the important information from the image. This results in a reduction in overall classification accuracy. The goal of the present research is to improve the image classification performance by combining the deep features extracted using popular deep convolutional neural network, VGG19, and various handcrafted feature extraction methods, i.e., SIFT, SURF, ORB, and Shi-Tomasi corner detector algorithm. Further, the extracted features from these methods are classified using various machine learning classification methods, i.e., Gaussian Naïve Bayes, Decision Tree, Random Forest, and eXtreme Gradient Boosting (XGBClassifier) classifier. The experiment is carried out on a benchmark dataset Caltech-101. The experimental results indicate that Random Forest using the combined features give 93.73% accuracy and outperforms other classifiers and methods proposed by other authors. The paper concludes that a single feature extractor whether shallow or deep is not enough to achieve satisfactory results. So, a combined approach using deep learning features and traditional handcrafted features is better for image classification.

Evaluating the Incidence of Audiological Derangement in Cases of Thyroid Hormone Imbalance.

Thyroid hormone disorders are both congenital as well as acquired. According to a projection from several studies on thyroid diseases, it has been estimated that about 42 million people in India suffer from various kind of thyroid diseases (1) Thyroid hormone is closely associated with the development of the cognitive functions, thus deficiency of this hormone is said to have effect an overall impact on proper functioning of the body. The normal functioning of the thyroid gland and adequate levels in the blood are Needed for the formation and functioning of the middle ear, inner ear and the central auditory pathway. Thus, congenital hypothyroidism (CH) can be a potential risk factor for hearing impairment (2) if the hormones decrease or are absent during the development of the peripheral and central auditory system structures. This study was undertaken with the objective of studying the pattern of hearing loss in patients who have a deranged thyroid profile. The study was conducted among 50 patients who were known cases of thyroid disorder in the Otorhinolaryngology Department of our institute. The study was hospital based observational clinical study. The patients were subjected to thyroid profile test After this, those who satisfied the inclusion and exclusion criteria and were included, after detailed history and examination, patients were subjected to PTA, hearing loss was classified according to WHO guidelines. The age group of the patients was from 30-55 yrs. with the mean age being 42. On the basis of the t3, t4 and tsh levels, in the current study out of 50 patients,40 were hypothyroid (80%),with male to female ratio of 6:4. On pure tone audiometry, 15 patients had decreased hearing. The other 25 had normal hearing. The incidence of hearing loss in hypothyoroid patients in our study is 37.5%. On analaysis the PTA reports of these patients,9 patients (22.5%) had conductive hearing loss of mild variety, the mean value of hearing loss being 26.2 decibels. 2 (5%) patients had mixed variety of hearing loss, with sensorineural hearing loss at higher frequencies. All the remaining patients had sensorineural type of hearing loss, i.e., 10%. Of the 10 patients, that have hyperthyroidism 8 were female and 2 were male. 3 patients (30%) had hearing loss, all these 3 patients reported hearing losses at high frequencies and had moderate type of sensorineural hearing loss. Conclusion- From our current study, we could see that hearing loss is present in both extremes of thyroid hormone imbalance The dominant variety of hearing loss in thyroid disorder is sensorineural type, because thyroid hormone imbalance affects the inner ear function.

Horizontal bone augmentation using two membranes at dehisced implant sites: A randomized clinical study.

Background: Placement of dental implant in narrow alveolar ridge is challenging to be treated. GBR procedure is currently most widely used to augment the deficient alveolar ridges and to treat the fenestration and dehiscence around dental implant. Thus, the objectives of the present study were to evaluate as well as compare the clinical performance of collagen membrane and titanium mesh for horizontal bone augmentation at dehisced implant sites. Methods and material: Total 12 single edentulous implant sites with buccal bone deficiency in 8 subjects were equally divided and treated simultaneously with either of the two membranes and DBBM(Bio-Oss) bone graft. Primary outcome measurements in terms of defect height and defect width were made using calibrated plastic periodontal probe. Re-entry surgery was performed to remeasure augmented site and to remove Ti-mesh at 6th months. Independent paired t-test for the inter-group comparison and student paired t-test for intra-group comparison were performed. The differences were considered to be significant at p ≤ 0.05. Results: Mean defect fill with respect to height and width was 3.50 ± 0.54 mm (87%) and 2.33 ± 0.51 mm (82%) for collagen membrane and 3.83 ± 0.75 mm (92%) and 2.50 ± 0.54 mm (88%) for Ti-mesh group respectively. Conclusions: Within the limitation of the study, it was concluded that mean defect height and width after 6 months were statistically significant within the group without significant difference between them, Although defect resolution was better in Ti-mesh.

Wet Versus Dry Temporalis Fascia Graft in Type 1 Tympanoplasty: A Randomized Study on Controversy.

Aim-To evaluate and compare the success rate of dry and wet temporalis fascia graft in tympanoplasty type I using an underlay technique and determine the effect of fibroblasts, if any on the success rate. Materials and methods-. STUDY DESIGN: A 3 year prospective, randomized, comparative study between 2019 and 2021. SETTING: Mahatma Gandhi University of Medical Sciences and Technology, Jaipur. SUBJECTS: 100 adult patients of either sex were taken in this study, ranging between 15 and 60 years who fulfilled inclusion and exclusion criteria. These patients were randomly divided into 2 groups of 50 each, one in dry graft and another in wet graft group. METHODS: A comparative analysis was done on the graft placement time, fibroblast count and hearing improvement postoperatively between the two groups. Results- The dry graft and wet graft groups had overall surgical success rates of 86 and 94 per cent, respectively; this finding was not statistically significant. A statistically significant high fibroblast count was observed in wet grafts, but it did not correlate with surgical success. Conclusion- A dry or wet temporalis fascia graft has no influence on the outcome of tympanoplasty type I.

Relation Between Posterior Canal Benign Paroxysmal Positional Vertigo and Vitamin D Deficiency.

The aim of this study is to establish a relationship between vitamin D levels and benign paroxysmal positional vertigo (BPPV) and its recurrence; and to evaluate the remedial effect of vitamin D on BPPV. In a pilot study, 40 patients with clinically diagnosed posterior canal BPPV were evaluated, treated and were followed up for a period of 6 months at ENT Department of Mahatma Gandhi Medical College, Jaipur. All the patients with clinical diagnosis of posterior canal BPPV were tested for 25(OH)D levels at their 1st visit. Patients were divided in two groups after appropriate matching. Group A was given vitamin D supplementation along with canal repositioning manoeuvre (CRM) and group B were only treated by canal repositioning manoeuvre. Patients underwent neuro-otological assessment during follow up visits and recurrent attacks were recorded. This study was conducted on 40 patients with BPPV (26 females and 14 males). The average age was 50 years. Average serum of 25(OH)D at the first visit was (12.2 ng/ml) for group A, and (12.4 ng/ml) for group B, all patients had low serum level of 25(OH)D (below 20 ng/ml). Recurrent BPPV episodes, were significantly lower in group A than that of group B. We second the hypothesis that the patients with BPPV who have insufficient levels of Vitamin D may benefit from vitamin D supplementation and we suggest further epidemiological and pharmacological studies to detect the effect of vitamin D deficiency on development and recurrence of vertigo.

Selective COX-2 Inhibitors: Road from Success to Controversy and the Quest for Repurposing.

The introduction of selective COX-2 inhibitors (so-called 'coxibs') has demonstrated tremendous commercial success due to their claimed lower potential of serious gastrointestinal adverse effects than traditional NSAIDs. However, following the repeated questioning on safety concerns, the coxibs 'controversial me-too' saga increased substantially, inferring to the risk of cardiovascular complications, subsequently leading to the voluntary withdrawal of coxibs (e.g., rofecoxib and valdecoxib) from the market. For instance, the makers (Pfizer and Merck) had to allegedly settle individual claims of cardiovascular hazards from celecoxib and valdecoxib. Undoubtedly, the lessons drawn from this saga revealed the flaws in drug surveillance and regulation, and taught science to pursue a more integrated translational approach for data acquisition and interpretation, prompting science-based strategies of risk avoidance in order to sustain the value of such drugs, rather than their withdrawal. Looking forward, coxibs are now being studied for repurposing, given their possible implications in the management of a myriad of diseases, including cancer, epilepsy, psychiatric disorders, obesity, Alzheimer's disease, and so on. This article briefly summarizes the development of COX-2 inhibitors to their market impression, followed by the controversy related to their toxicity. In addition, the events recollected in hindsight (the past lessons), the optimistic step towards drug repurposing (the present), and the potential for forthcoming success (the future) are also discussed.

The Genus Alternanthera: Phytochemical and Ethnopharmacological Perspectives.

Ethnopharmacological relevance: The genus Alternanthera (Amaranthaceae) comprises 139 species including 14 species used traditionally for the treatment of various ailments such as hypertension, pain, inflammation, diabetes, cancer, microbial and mental disorders. Aim of the review: To search research gaps through critical assessment of pharmacological activities not performed to validate traditional claims of various species of Alternanthera. This review will aid natural product researchers in identifying Alternanthera species with therapeutic potential for future investigation. Materials and methods: Scattered raw data on ethnopharmacological, morphological, phytochemical, pharmacological, toxicological, and clinical studies of various species of the genus Alternanthera have been compiled utilizing search engines like SciFinder, Google Scholar, PubMed, Science Direct, and Open J-Gate for 100 years up to April 2021. Results: Few species of Alternanthera genus have been exhaustively investigated phytochemically, and about 129 chemical constituents related to different classes such as flavonoids, steroids, saponins, alkaloids, triterpenoids, glycosides, and phenolic compounds have been isolated from 9 species. Anticancer, antioxidant, antibacterial, CNS depressive, antidiabetic, analgesic, anti-inflammatory, and immunomodulator effects have been explored in the twelve species of the genus. A toxicity study has been conducted on 3 species and a clinical study on 2 species. Conclusions: The available literature on pharmacological studies of Alternanthera species reveals that few species have been selected based on ethnobotanical surveys for scientific validation of their traditional claims. But most of these studies have been conducted on uncharacterized and non-standardized crude extracts. A roadmap of research needs to be developed for the isolation of new bioactive compounds from Alternanthera species, which can emerge out as clinically potential medicines.

Novel Therapies for the Treatment of Neuropathic Pain: Potential and Pitfalls.

Neuropathic pain affects more than one million people across the globe. The quality of life of people suffering from neuropathic pain has been considerably declining due to the unavailability of appropriate therapeutics. Currently, available treatment options can only treat patients symptomatically, but they are associated with severe adverse side effects and the development of tolerance over prolonged use. In the past decade, researchers were able to gain a better understanding of the mechanisms involved in neuropathic pain; thus, continuous efforts are evident, aiming to develop novel interventions with better efficacy instead of symptomatic treatment. The current review discusses the latest interventional strategies used in the treatment and management of neuropathic pain. This review also provides insights into the present scenario of pain research, particularly various interventional techniques such as spinal cord stimulation, steroid injection, neural blockade, transcranial/epidural stimulation, deep brain stimulation, percutaneous electrical nerve stimulation, neuroablative procedures, opto/chemogenetics, gene therapy, etc. In a nutshell, most of the above techniques are at preclinical stage and facing difficulty in translation to clinical studies due to the non-availability of appropriate methodologies. Therefore, continuing research on these interventional strategies may help in the development of promising novel therapies that can improve the quality of life of patients suffering from neuropathic pain.

Comparative assessment of conventional periodontal probes and CEJ handpiece of electronic probes in the diagnosis and primary care of periodontal disease.

OBJECTIVES: Conventional probes (CPs) have been considered acceptable as diagnostic tools to measure probing pocket depth (PPD) and clinical attachment level (CAL) but are affected by multiple variables. Electronic probes (EPs) provide controlled force, digital readout and data storage in computers. The objectives were to compare the reproducibility in the measurement of PPD and CAL by CP and the newly introduced CEJ handpiece of EP and intra-examiner and inter-examiner errors done in two phases. METHODS AND MATERIAL: Selected 720 periodontal sites in 1st molar of 30 persons with chronic periodontitis ≤4 mm and >4 mm pockets were analysed by two trained investigators in two phases at 2 hours difference by CP and CEJ handpiece of EP. Standard deviation, mean difference, correlation coefficient, P value and student 't' test were done to analyse data. RESULTS: The intra- examiner and inter-examiner analyses revealed that Pearson's correlation coefficient was above 0.080 and 0.722 in the ≤4 mm and >4 mm pockets, respectively. Mean difference was not statistically significant in both groups except in the intra- examiner findings in the 2nd phase. Interprobe analysis depicted a standard error of mean of <0.03 in ≤4 mm pockets, whereas it varied from 0.047-0.056 in >4 mm pockets. CONCLUSION: In conclusion, EP is advantageous for research purposes by providing automatic recording and long-term maintenance of data storage without the need of an assistant and patient education and motivation, whereas CP appears to be more useful in routine periodontal examination.

Neurological Consequences of SARS-CoV-2 Infection and Concurrence of Treatment-Induced Neuropsychiatric Adverse Events in COVID-19 Patients: Navigating the Uncharted.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) binds to the angiotensin-converting enzyme 2 (ACE2) receptor and invade the human cells to cause COVID-19-related pneumonia. Despite an emphasis on respiratory complications, the evidence of neurological manifestations of SARS-CoV-2 infection is rapidly growing, which is substantially contributing to morbidity and mortality. The neurological disorders associated with COVID-19 may have several pathophysiological underpinnings, which are yet to be explored. Hypothetically, SARS-CoV-2 may affect the central nervous system (CNS) either by direct mechanisms like neuronal retrograde dissemination and hematogenous dissemination, or via indirect pathways. CNS complications associated with COVID-19 include encephalitis, acute necrotizing encephalopathy, diffuse leukoencephalopathy, stroke (both ischemic and hemorrhagic), venous sinus thrombosis, meningitis, and neuroleptic malignant syndrome. These may result from different mechanisms, including direct virus infection of the CNS, virus-induced hyper-inflammatory states, and post-infection immune responses. On the other hand, the Guillain-Barre syndrome, hyposmia, hypogeusia, and myopathy are the outcomes of peripheral nervous system injury. Although the therapeutic potential of certain repurposed drugs has led to their off-label use against COVID-19, such as anti-retroviral drugs (remdesivir, favipiravir, and lopinavir-ritonavir combination), biologics (tocilizumab), antibiotics (azithromycin), antiparasitics (chloroquine and hydroxychloroquine), and corticosteroids (dexamethasone), unfortunately, the associated clinical neuropsychiatric adverse events remains a critical issue. Therefore, COVID-19 represents a major threat to the field of neuropsychiatry, as both the virus and the potential therapies may induce neurologic as well as psychiatric disorders. Notably, potential COVID-19 medications may also interact with the medications of pre-existing neuropsychiatric diseases, thereby further complicating the condition. From this perspective, this review will discuss the possible neurological manifestations and sequelae of SARS-CoV-2 infection with emphasis on the probable underlying neurotropic mechanisms. Additionally, we will highlight the concurrence of COVID-19 treatment-associated neuropsychiatric events and possible clinically relevant drug interactions, to provide a useful framework and help researchers, especially the neurologists in understanding the neurologic facets of the ongoing pandemic to control the morbidity and mortality.

Integrating CRISPR-Cas and Next Generation Sequencing in Plant Virology.

Plant pathology has been revolutionized by the emergence and intervention of next-generation sequencing technologies (NGS) which provide a fast, cost-effective, and reliable diagnostic for any class of pathogens. NGS has made tremendous advancements in the area of research and diagnostics of plant infecting viromes and has bridged plant virology with other advanced research fields like genome editing technologies. NGS in a broader perspective holds the potential for plant health improvement by diagnosing and mitigating the new or unusual symptoms caused by novel/unidentified viruses. CRISPR-based genome editing technologies can enable rapid engineering of efficient viral/viroid resistance by directly targeting specific nucleotide sites of plant viruses and viroids. Critical genes such as eIf (iso) 4E or eIF4E have been targeted via the CRISPR platform to produce plants resistant to single-stranded RNA (ssRNA) viruses. CRISPR/Cas-based multi-target DNA or RNA tests can be used for rapid and accurate diagnostic assays for plant viruses and viroids. Integrating NGS with CRISPR-based genome editing technologies may lead to a paradigm shift in combating deadly disease-causing plant viruses/viroids at the genomic level. Furthermore, the newly discovered CRISPR/Cas13 system has unprecedented potential in plant viroid diagnostics and interference. In this review, we have highlighted the application and importance of sequencing technologies on covering the viral genomes for precise modulations. This review also provides a snapshot vision of emerging developments in NGS technologies for the characterization of plant viruses and their potential utilities, advantages, and limitations in plant viral diagnostics. Furthermore, some of the notable advances like novel virus-inducible CRISPR/Cas9 system that confers virus resistance with no off-target effects have been discussed.

Exophytic gingival growth of the maxillary canine region in a young individual: Extremely rare case report of peripheral dentinogenic ghost cell tumor.

The present case report is a case of peripheral dentinogenic ghost cell tumor (PDGCT), an extremely rare solid benign neoplastic variant of calcifying cystic odontogenic tumor of the gingiva mimicking clinically as pyogenic granuloma, fibroma, peripheral ossifying fibroma, and peripheral giant-cell granuloma. A 24-year-old male reported with painless, firm, solitary, sessile, smooth-surfaced, nonulcerative, nonpulsatile, well-defined swelling measuring ≈12 mm × 9 mm in the interdental gingiva of the teeth #13 and #14 extending to the mucogingival junction. Intraoral periapical radiographic showed a normal trabecular pattern with mild radiolucency without bony expansion, periapical lesion, and resorption of the adjacent teeth. The diagnosis was established by histopathologic examination. Very few cases of this entity have been documented in the literature. The present case report aims to document this rare entity and emphasizes on the fact that histopathological examination of every localized gingival growth should be included in the treatment planning to differentiate with other commonly found lesions.

Clinical and anorectal manometry profile of patients with functional constipation and constipation-predominant irritable bowel syndrome.

BACKGROUND: Functional constipation (FC) and constipation-predominant irritable bowel syndrome (IBS-C) have significant healthcare impact. Clinical and investigative data of patients with these disorders in Indian population is scarce. We aimed to compare the clinical and anorectal manometric profile of patients with FC and IBS-C. METHODS: Consecutive patients with chronic constipation undergoing anorectal manometry (ARM) and balloon expulsion test (BET) were enrolled. Thirty healthy volunteers served as controls (HC). Functional defecatory disorder (FDD) was diagnosed according to ROME IV criteria if both ARM and BET were abnormal. RESULTS: Of the 231 patients enrolled (median age 47 years, 87.8% males), FC and IBS-C were diagnosed in 132 (57.1%) and 99 (42.9%) patients, respectively. Significant clinical differences between FC and IBS-C patients included older age, lower stool frequency/week, higher frequency of straining, and greater frequency  of incomplete evacuation (p < 0.001). ARM revealed abnormal defecatory pattern in 55.3% (n = 73) FC patients and 47.5% (n = 47) IBS-C patients. Of them, 54.7% (40/73) of FC patients had inadequate defecatory propulsion while 89.4% (42/47) of the IBS-C patients had dyssynergic defecation (p < 0.001). Abnormal BET was seen in 67.4% of FC patients and 43.4% of IBS-C patients. Thus, FDD was diagnosed in 55.3% and 46.5% of FC and IBS-C patients, respectively. Rectal hyposensitivity was present in 60.6% of FC patients compared with 2% of IBS-C patients (p < 0.001). CONCLUSIONS: There were significant differences in clinical and manometric profile of FC and IBS-C patients. Compared with IBS-C patients, FC patients were older, had higher prevalence of FDD, less often had dyssynergic defecation, and had higher prevalence of rectal hyposensitivity.