3Bs of CRISPR-Cas mediated genome editing in plants: exploring the basics, bioinformatics and biosafety landscape.

The recent thrust in research has projected the type II clustered regularly interspaced short palindromic repeats and associated protein 9 (CRISPR-Cas9) system as an avant-garde plant genome editing tool. It facilitates the induction of site-specific double-stranded DNA cleavage by the RNA-guided DNA endonuclease (RGEN), Cas9. Elimination, addition, or alteration of sections in DNA sequence besides the creation of a knockout genotype (CRISPRko) is aided by the CRISPR-Cas9 system in its wild form (wtCas9). The inactivation of the nuclease domain generates a dead Cas9 (dCas9), which is capable of targeting genomic DNA without scissoring it. The dCas9 system can be engineered by fusing it with different effectors to facilitate transcriptional activation (CRISPRa) and transcriptional interference (CRISPRi). CRISPR-Cas thus holds tremendous prospects as a genome-manipulating stratagem for a wide gamut of crops. In this article, we present a brief on the fundamentals and the general workflow of the CRISPR-Cas system followed by an overview of the prospects of bioinformatics in propelling CRISPR-Cas research with a special thrust on the available databases and algorithms/web-accessible applications that have aided in increasing the usage and efficiency of editing. The article also provides an update on the current regulatory landscape in different countries on the CRISPR-Cas edited plants to emphasize the far-reaching impact of the genomic editing technology. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01397-3.

Psychodynamic Perspective of Sexual Obsessions in Obsessive-Compulsive Disorder.

Obsessive-compulsive disorder (OCD) is a disorder that is characterized by the presence of obsessions and compulsions. OCD can occur in a wide range of subtypes, which may include sexual OCD, i.e., the occurrence of intrusive sexual thoughts. People with sexual OCD may experience unwanted obsessive sexual thoughts about a family member, dead or inanimate objects, animals, God, or children, which can range from mild to severe levels of occurrence. The psychodynamic perspective on OCD suggests that the content of obsessive thinking is commonly drawn from primitivized sexuality and aggression. It further suggests that in the absence of key relationships that include emotional proximity, mirroring and containment, and attunement, a child experiences a void-like state. Consequently, the anxiety that arises in the child leads to a form of liveliness in a "dead" inner world. However, it occurs out of a sense of abandonment and loss of good objects, and ultimately out of fear of annihilation. As a result, the child resorts to maladaptive defense mechanisms. The study attempts to explore the adverse early childhood experiences such as parental neglect, mother-father conflict, and lack of parental love and care leading to fixation in psychosexual stages of development, which further leads to the adoption of maladaptive defense mechanisms that in turn further contribute to the development of obsessional personality traits. This study aims to explain the psychodynamic perspective on the emergence of sexual obsessions by proceeding with a case study approach by taking a diagnosed case of OCD. A comprehensive psychodiagnostic assessment was conducted with the help of Draw-a-Person Test, Thematic Apperception Test, Rotter's Incomplete Sentence Blank, and Million's Comprehensive Multiaxial Inventory. The analysis of the assessment protocol revealed a strong relationship between the defense mechanism, personality traits, and the development of obsessive symptoms.

Effectiveness of Neurofeedback Therapy Adjunct to Cognitive Behavioral Therapy in Agoraphobia: A Case Study.

Agoraphobia has been defined as marked fear or anxiety being in enclosed or open places, using public transportation, and being in a crowd or outside of home alone. Such individuals make active attempts to avoid those places which cause intense distress. The neuronal areas that play a significant role in agoraphobia are uncinate fasciculus, which connects the prefrontal lobe and amygdala and various alterations in anterior cingulate cortex, insula, amygdala, and lateral prefrontal cortex. Neurofeedback is a type of biofeedback which teaches self-control of brain functions by measuring brain waves with the help of electroencephalography (EEG) and providing a feedback signal. With the help of alpha and beta training protocol, neurofeedback therapy will help in enhancing the connectivity between prefrontal cortex and amygdala. The present study aims to explore the therapeutic effectiveness of neurofeedback therapy as an adjunct treatment with cognitive behavioral therapy (CBT) in patients with agoraphobia disorder. A single case study method was adopted. The patient having the symptoms of agoraphobia disorder diagnosed as per ICD-10 criteria was taken in the study. After going through detailed case history and mental status examination, the patient was assessed on psychological measures on baseline and other follow-up visits. A total of 18 therapeutic sessions of neurofeedback therapy (alpha and beta protocol) along with CBT were conducted. Intermittent assessments on Draw A Person Test (DAPT), EEG parameters, Visual Analogue Scale (VAS), and Panic and Agoraphobia Scale (PAS) were conducted to compare the pre- and post-assessment findings. The results indicated significant improvement in the symptoms of the patient after intervention. The pre- and post-assessment findings and the neurofeedback therapy along with CBT were observed to be effective in treating the symptoms of agoraphobia. Neurofeedback therapy and CBT were proven to be effective in removing the symptoms of agoraphobia disorder within the patient.

Sequence based prediction of enhancer regions from DNA random walk.

Regulatory elements play a critical role in development process of eukaryotic organisms by controlling the spatio-temporal pattern of gene expression. Enhancer is one of these elements which contributes to the regulation of gene expression through chromatin loop or eRNA expression. Experimental identification of a novel enhancer is a costly exercise, due to which there is an interest in computational approaches to predict enhancer regions in a genome. Existing computational approaches to achieve this goal have primarily been based on training of high-throughput data such as transcription factor binding sites (TFBS), DNA methylation, and histone modification marks etc. On the other hand, purely sequence based approaches to predict enhancer regions are promising as they are not biased by the complexity or context specificity of such datasets. In sequence based approaches, machine learning models are either directly trained on sequences or sequence features, to classify sequences as enhancers or non-enhancers. In this paper, we derived statistical and nonlinear dynamic features along with k-mer features from experimentally validated sequences taken from Vista Enhancer Browser through random walk model and applied different machine learning based methods to predict whether an input test sequence is enhancer or not. Experimental results demonstrate the success of proposed model based on Ensemble method with area under curve (AUC) 0.86, 0.89, and 0.87 in B cells, T cells, and Natural killer cells for histone marks dataset.

HIV patient with mucous membrane pemphigoid: a case report.

BACKGROUND: Mucous Membrane Pemphigoid, a new denomination of cicatricial pemphigoid, encompasses a group of chronic subepithelial autoimmune blistering diseases that predominantly affect the oral cavity and the eyes (conjunctivitis and symblepharon). CASE DETAILS: A rare case of Mucous Membrane Pemphigoid (MMP) in a human immunodeficiency virus (HIV) positive patient is discussed with clinicohistopathological presentation. CONCLUSION: Since our patient was HIV-positive and had lesions restricted to the oral mucosa with ocular involvement, only topical and intralesional steroids were preferred as the first line of treatment. Systemic corticosteroid therapy raises a concern regarding immunosuppression.

Estimation of serum ß2-microglobulin in potentially malignant disorders and squamous cell carcinoma of the oral cavity: A clinicopathological study.

BACKGROUND: Tumor markers are substances, which quantitatively changes in serum, during the tumor development, one such tumor marker is serum ß2-microglobulin (ß2-m). The aim of this study was to establish the role of ß2-m as a biochemical parameter for diagnosis and prognosis of oral carcinoma by estimation of serum ß2-m levels in potentially malignant lesions, conditions, and oral squamous cell carcinoma. MATERIALS AND METHODS: The study was carried out on 48 subjects (16 control, 8 oral submucous fibrosis, 8 oral leukoplakia, and 16 oral squamous cell carcinoma patients of different stages), conducted at department of Oral Medicine, Kothiwal Dental College, Moradabad, India. Under aseptic precautions, 5 ml venous blood was drawn and serum was separated. Estimation of ß2-m level in serum was carried out by enzyme linked immunosorbent assay. The data were analyzed by using the statistical package for social sciences (SPSS 17.0) software. Cases and controls were tested for statistical significance with one-way ANOVA with post-hoc Tukey's HSD. Values of P < 0.05 were considered significant. RESULTS: The mean serum ß2-m level in the control group was 1.173 ± 0.059, in potentially malignant lesions/conditions group was 1.688 ± 0.137 and in oral squamous cell carcinoma group was 2.835 ± 0.0313. This progressive increase in serum ß2-m level was found to be highly significant (P value < 0.001). Results of Receiver operating characteristic analysis showed ß2-m as a 100% sensitive and specific biomarker for oral squamous cell carcinoma. CONCLUSION: The present study establishes ß2-m as a specific biological tumor marker for diagnostic and prognostic evaluation of oral squamous cell carcinoma.

Dual-color fluorescence cross-correlation spectroscopy on a single plane illumination microscope (SPIM-FCCS).

Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, flow velocities and concentrations in an imaging mode. Here we extend this technique to two-color fluorescence cross-correlation spectroscopy (SPIM-FCCS), which allows to measure molecular interactions in an imaging mode. We present a theoretical framework for SPIM-FCCS fitting models, which is subsequently used to evaluate several test measurements of in-vitro (labeled microspheres, several DNAs and small unilamellar vesicles) and in-vivo samples (dimeric and monomeric dual-color fluorescent proteins, as well as membrane bound proteins). Our method yields the same quantitative results as the well-established confocal FCCS, but in addition provides unmatched statistics and true imaging capabilities.

Adenolymphoma: A probing entity: Case report and review.

Warthin's tumor undoubtedly is the most frequent monomorphic adenoma of the major salivary glands. Clinically, it appears as a slow-growing tumor often fluctuant on palpation due to its cystic nature. The treatment of choice is complete excision with wide tumor-free margins. This article highlights a case of Warthin's tumor of the parotid gland in an elderly male patient along with a review of the literature on the aforementioned pathology.

The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy.

Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, transport, flow velocities and concentrations in an imaging mode. SPIM-FCS records correlation functions over a whole plane in a sample, which requires array detectors for recording the fluorescence signal. Several types of image sensors are suitable for FCS. They differ in properties such as effective area per pixel, quantum efficiency, noise level and read-out speed. Here we compare the performance of several low light array detectors based on three different technologies: (1) Single-photon avalanche diode (SPAD) arrays, (2) passive-pixel electron multiplying charge coupled device (EMCCD) and (3) active-pixel scientific-grade complementary metal oxide semiconductor cameras (sCMOS). We discuss the influence of the detector characteristics on the effective FCS observation volume, and demonstrate that light sheet based SPIM-FCS provides absolute diffusion coefficients. This is verified by parallel measurements with confocal FCS, single particle tracking (SPT), and the determination of concentration gradients in space and time. While EMCCD cameras have a temporal resolution in the millisecond range, sCMOS cameras and SPAD arrays can extend the time resolution of SPIM-FCS down to 10 µs or lower.

Initial expression of the Papillon-Lefevre syndrome in consanguine family.

A rare case of Papillon-Lefevre syndrome is discussed with clinicoradiological presentation. The purpose of the case report is to make the medical community aware of this rare syndrome and its association with consanguinity. Papillon-Lefevre syndrome is an extremely rare genodermatosis of autosomal-recessive inheritance which usually manifests itself between the ages of 6 months to 4 years characterised by diffuse palmoplanter hyperkeratosis (keratoderma), and rapidly progressive and devastating periodontitis, affecting the primary as well as permanent dentition. Papillon-Lefevre syndrome results from a combination of host and bacterial factors, including recessive gene, consanguinity, specific periodontal pathogens and lack of thorough oral hygiene. The present case report describes Papillon-Lefevre syndrome and its association with consanguinity in a 3-year-old girl.

A rare presentation of dens in dente in the mandibular third molar with extra oral sinus.

The unusual case of dense in dente in mandibular molar area with extra oral sinus in a 30-year-old female is presented. The chief complaint of the patient was wound formation and pus discharge from the right side of lower jaw for many years. Clinical examination revealed extra oral sinus and mild swelling in vestibular region opposite the right mandibular molars. On radiographical examination, right mandibular third molar had bulbous root associated with periapical radiolucency. It appeared that there was a tooth within a tooth and the invagination extended nearly to the root apex. A clinical diagnosis of dense in dente Type III was confirmed by stereomicroscopy of ground section of the tooth. Extra oral sinus healed after extraction of the mandibular third molar within a month.

Cognitive functions in euthymic state of bipolar disorder: an Indian study.

INTRODUCTION: It has recently been observed that some cognitive deficits in bipolar disorders persist even after the subsidence of active symptoms. The authors aim to study the cognitive functioning of patients with bipolar disorder, currently in euthymia and compare them with normal healthy controls. METHODS: Fifteen patients having bipolar-I disorder and currently in euthymia were compared with fifteen age- and education-matched controlled subjects. Cognitive assessments were done using three computer-based tests, i.e., Wisconsin's Card Sorting Test (WCST), Spatial Working Memory Test (SWMT), and Continuous Performance Test (CPT). RESULTS: Euthymic bipolar patients showed significant deficits in executive functions. Subtle deficits were present in attention and working memory that were not statistically significant. CONCLUSIONS: Executive deficits may be trait markers in bipolar disorder and may have clinical implications in patient rehabilitation.

Executive functions in depression: a clinical report.

Fifty patients of depression and thirty normal subjects were assessed using clinical rating scales and also for the executive functions by Wisconsin Card Sorting Test (WCST). The depressed subjects demonstrated poor performance on WCST suggesting cognitive inflexibility and prefrontal dysfunction. More severe illness was associated with greater impairment in the executive functioning. This pattern of result in conjunction with previous studies supported the idea that depressed patients may have fixed frontally based dysfunction and calls for the use of cognitive assessment and rehabilitation in the patients with depression.