Expression of circulating tumour cells in oral squamous cell carcinoma: An ex vivo pilot study.

Introduction: Despite advances in diagnostics and therapeutics, two-thirds of oral cancer patients present with advanced disease, which increases both the morbidity and mortality risk. Circulating tumour cells (CTCs) are released in the circulation by primary tumours and have been demonstrated to have significant correlations between their occurrence and disease progression. Objectives: To characterize the circulating tumour cells in subjects with histologically diagnosed oral squamous cell carcinoma (OSCC). Materials and Methods: This pilot study was undertaken with ten fresh blood samples (6 ml each). Five samples from apparently healthy individuals and five OSCC samples were cultured and subjected to flow cytometric analysis for CD44 expression. Immunostaining was done using CD44 and EpCAM markers. Result: Several cells in OSCC samples showed EpCAM and CD44 positivity following immunostaining. However, flow cytometry performed with CD44 alone was not specific for OSCC samples. Hence, proving that CD44 and EpCAM when used in conjunction can help to characterize CTCs. Conclusion: The findings of our study suggest that the demonstration of CTCs is feasible and helps in understanding of disease progression and metastatic risk. Sensitive detection of CTCs from blood samples can serve as an implicit tool in early cancer diagnosis and prognosis through liquid biopsy which in itself is minimally invasive and time-saving.

Expression of PTCH gene in ameloblastoma and odontogenic keratocyst: A comparative study.

Aim: This is a cross-sectional comparative study, aimed to quantify the expression of patched (PTCH) gene in ameloblastoma, odontogenic keratocyst (OKC) and also the comparison of both the expressions. Materials and Methods: Genomic deoxyribonucleic acid (DNA) was extracted and quantified, and the expression of the PTCH gene was done in 17 cases of ameloblastoma and 17 cases of OKC by quantitative real-time polymerase chain reaction (RT-qPCR). Results: It was observed that there was an overexpression of the PTCH gene in both ameloblastoma and OKC with a good mean cycle threshold (CT) value of 32.71 ± 2.432 and 34.69 ± 1.875, respectively. When comparing the PTCH expression between the two, ameloblastoma showed higher expression than the OKC and the difference is statistically significant with P value of 0.025. Conclusions: Our findings suggest that there is overexpression of PTCH in ameloblastoma and OKC, but it is highly expressed in ameloblastoma when compared to OKC. Overexpression of PTCH may constitute the activation of the Sonic Hedgehog pathway and may suggest the mechanism for the development of ameloblastoma and OKC. Hence it can be used as a valuable marker for early diagnosis and in the identification of therapeutic targets.

Rice-Magnaporthe transcriptomics reveals host defense activation induced by red seaweed-biostimulant in rice plants.

Red seaweed extracts have been shown to trigger the biotic stress tolerance in several crops. However, reports on transcriptional modifications in plants treated with seaweed biostimulant are limited. To understand the specific response of rice to blast disease in seaweed-biostimulant-primed and non-primed plants, transcriptomics of a susceptible rice cultivar IR-64 was carried out at zero and 48 h post inoculation with Magnaporthe oryzae (strain MG-01). A total of 3498 differentially expressed genes (DEGs) were identified; 1116 DEGs were explicitly regulated in pathogen-inoculated treatments. Functional analysis showed that most DEGs were involved in metabolism, transport, signaling, and defense. In a glass house, artificial inoculation of MG-01 on seaweed-primed plants resulted in the restricted spread of the pathogen leading to the confined blast disease lesions, primarily attributed to reactive oxygen species (ROS) accumulation. The DEGs in the primed plants were defense-related transcription factors, kinases, pathogenesis-related genes, peroxidases, and growth-related genes. The beta-D-xylosidase, a putative gene that helps in secondary cell wall reinforcement, was downregulated in non-primed plants, whereas it upregulated in the primed plants indicating its role in the host defense. Additionally, Phenylalanine ammonia-lyase, pathogenesis-related Bet-v-I family protein, chalcone synthase, chitinases, WRKY, AP2/ERF, and MYB families were upregulated in seaweed and challenge inoculated rice plants. Thus, our study shows that priming rice plants with seaweed bio-stimulants resulted in the induction of the defense in rice against blast disease. This phenomenon is contributed to early protection through ROS, protein kinase, accumulation of secondary metabolites, and cell wall strengthening.

Insights on Novel Effectors and Characterization of Metacaspase (RS107_6) as a Potential Cell Death-Inducing Protein in Rhizoctonia solani.

Effectors play an important role in host-pathogen interactions. Though an economically significant disease in rice, knowledge regarding the infection strategy of Rhizoctonia solani is obscure. In this study, we performed a genome-wide identification of the effectors in R. solani based on the characteristics of previously reported effector proteins. A total of seven novel effectors (designated as RS107_1 to RS107_7) in the disease mechanism of R. solani were identified and were predicted to be non-classically secreted proteins with functionally conserved domains. The function, reactivity, and stability of these proteins were evaluated through physiochemical characterization. The target proteins involved in the regulation of rice defense mechanisms were identified. Furthermore, the effector genes were cloned and RS107_6 (metacaspase) was heterologously expressed in Escherichia coli to obtain a purified protein of ~36.5 kDa. The MALD-TOF characterization confirmed that the protein belonged to a metacaspase of the Peptidase_C14 protein family, 906 bp in size, and encoded a polypeptide of 301 amino acids. These findings suggest that the identified effectors can potentially serve as a virulence factor and can be targeted for the management of sheath blight in rice.

Bacillus velezensis (strains A6 & P42) as a potential biocontrol agent against Klebsiella variicola, a new causal agent of soft rot disease in carrot.

Bacterial soft rot is one of the most devastating diseases and a major constraint encountered during carrot farming. Biological agents are the best eco-friendly alternatives to agrochemicals to manage soft rot disease to ensure environmental sustainability. In this study, about eight isolates of bacterial pathogen causing soft rot in carrots were collected from Karnataka, India. Based on the 16S rRNA sequencing the pathogen isolates causing soft rot of carrot were identified as Klebsiella variicola. The morphological characteristics of K. variicola was investigated under scanning electron microscopy. The pathogenicity assay showed that all eight isolates were pathogenic to the carrot. An in vitro and in planta assay of two novel strains of Bacillus velezensis (A6 and P42) against K. variicola indicated that both strains had strong antagonistic activity against all the pathogen strains. Furthermore, the volatile bioactive compounds produced by A6 and P42 strains were analyzed in GC-MS, which revealed the presence of 10 and 6 bioactive compounds in their culture filtrate, respectively, with antibacterial and antifungal properties. The present study suggests that both A6 and P42 strains of B. velezensis were antagonistic to K. variicola and can be used as biocontrol agents to manage soft rot diseases of carrot under field conditions.

Hand Use and Grasp Sensor System in Monitoring Infant Fine Motor Development.

Objective: To assess the feasibility of a hand use and grasp sensor system in collecting and quantifying fine motor development longitudinally in an infant's home environment. Design: Cohort study. Researchers made home visits monthly to participating families to collect grasp data from infants using a hand use and grasp sensor. Setting: Data collection were conducted in each participant's home. Participants: A convenience sample of 14 typical developmental infants were enrolled from 3 months to 9 months of age. Two infants dropped out. A total of 62 testing sessions involving 12 infants were available for analysis (N=12). Interventions: At each session, the infant was seated in a standardized infant seat. Each instrumented toy was hung on the hand use and grasp sensor structure, presented for 6 minutes in 3 feedback modes: visual, auditory, and vibratory. Main Outcome Measures: Infant grasp frequency and duration, peak grasping force, average grasping force, force coefficient of variation, and proportion of bimanual grasps. Results: A total of 2832 recorded grasp events from 12 infants were analyzed. In linear mixed-effects model analysis, when interacting with each toy, infants' peak grasp force, average grasp force, and accumulated grasp time all increased significantly with age (all P<.001). Bimanual grasps also occupied an increasingly greater percentage of infants' total grasps as they grew older (bar toy P<.001, candy toy P=.021). Conclusions: We observed significant changes in hand use and grasp sensor outcome measures with age that are consistent with maturation of grasp skills. We envision the evolution of hand use and grasp sensor technology into an inexpensive and convenient tool to track infant grasp development for early detection of possible developmental delay and/or cerebral palsy as a supplement to clinical evaluations.

Red-seaweed biostimulants differentially alleviate the impact of fungicidal stress in rice (Oryza sativa L.).

Red seaweed-derived biostimulants facilitate plant health and impart protection against abiotic stress conditions by their bioactive compounds and plant nutrients. The potency of red seaweed biostimulants (LBS6 and LBD1) on rice cv. IR-64 in response to fungicides induced stress was investigated in this study. Foliar application of LBS6 maintained the stomatal opening and leaf temperature under the fungicidal stress condition. Reactive Oxygen Species (ROS) such as hydrogen peroxide and superoxide radicals were significantly reduced in LBS6-treated stressed plants. After applying seaweed biostimulants, ROS production was stabilized by antioxidants viz., CAT, APX, SOD, POD, and GR. LBS-6 application increased the Ca+ and K+ levels in the stressed plants, which perhaps interacted with ROS and stomatal opening signalling systems, respectively. In the rice plants, fungicidal stress elevated the expression of stress-responsive transcriptional factors (E2F, HSFA2A, HSFB2B, HSFB4C, HSFC1A, and ZIP12). A decline in the transcript levels of stress-responsive genes was recorded in seaweed treated plants. For the first time, we present an integrative investigation of physicochemical and molecular components to describe the mechanism by which seaweed biostimulants in rice improve plant health under fungicidal stress conditions.

Pyroptosis and pyroptosis-inducing cancer drugs.

Pyroptosis, an inflammatory form of lytic cell death, is a type of cell death mediated by the gasdermin (GSDM) protein family. Upon recognizing exogenous or endogenous signals, cells undergo inflammasome assembly, GSDM cleavage, the release of proinflammatory cytokines and other cellular contents, eventually leading to inflammatory cell death. In this review, we discuss the roles of the GSDM family for anti-cancer functions and various antitumor drugs that could activate the pyroptosis pathways.

Role of a dentist in early diagnosis of Granulomatosis with Polyangiitis: A rare case report.

Granulomatosis with polyangiitis (GPA) is a rare multisystem disease characterized by a triad of necrotizing granulomas affecting the upper and lower respiratory tract, vasculitis and glomerulonephritis. Oral lesions are seen in 6-13% of cases, although not life threatening but might be the first symptoms reported by the patient. Here we present a case of gingival growth, later diagnosed as GPA owing to the timely recognition by the dentist. A 33-year-old female reported for evaluation of soft tissue growth in her lower left back tooth region present for past 3 months. She had no relevant medical or family history. Clinically it looked like a benign hypertrophic gingival enlargement. The histopathological picture was suggestive of vasculitis and nonspecific granuloma formation. On further examination altered Renal Function Test and cavitating granulomas were noted in her lungs along with a positive cytoplasmic ANCA (c-ANCA) test which led to the diagnosis of GPA.

Rapid genotyping of bacterial leaf blight resistant genes of rice using loop-mediated isothermal amplification assay.

The use of resistant (R) genes is the most effective strategy to manage bacterial leaf blight (BLB) disease of rice. Several attempts were made to incorporate R genes into susceptible rice cultivars using marker-assisted backcross breeding (MABB). However, MABB relies exclusively on PCR for foreground selection of R genes, which requires expensive equipment for thermo-cycling and visualization of results; hence, it is limited to sophisticated research facilities. Isothermal nucleic acid amplification techniques such as loop-mediated isothermal amplification (LAMP) assay do not require thermo-cycling during the assay. Therefore, it will be the best alternative to PCR-based genotyping. In this study, we have developed a LAMP assay for the specific and sensitive genotyping of seven BLB resistance (R) genes viz., Xa1, Xa3, Xa4, Xa7, Xa10, Xa11, and Xa21 in rice. Gene-specific primers were designed for the LAMP assay. The LAMP assay was optimized for time, temperature, and template DNA concentration. For effective detection, incubation at 60 °C for 30 min was optimum for all seven R genes. A DNA intercalating dye ethidium bromide and a calorimetric dye hydroxynaphthol blue was used for result visualization. Further, sensitivity assay revealed that the LAMP assay could detect R genes at 100 fg of template DNA compared to 1 ng and 10 pg, respectively, in conventional PCR and q-PCR assays. The LAMP assay developed in this study provides a simple, specific, sensitive, robust, and cost-effective method for foreground selection of R genes in the resistance breeding programs of resource-poor laboratory.

Cysticercosis Masquerading as Lipo-Dermoid.

Cysticercosis is caused by the larval stage of Taenia solium (Pork tape worm). Humans are the definitive hosts where the life cycle is completed and pig is the intermediate host. Incidence in India varies from 2-38% based on the consumption of pork, general hygiene and socio-economic status of the population. The parasite on reaching the small intestine encysts itself in various tissues. Most commonly Brain, Eyes, subcutaneous tissue. Intravitreal cyst is the commonest intra ocular presentation. Death of the parasite and release of the contents of the cyst causes an intense inflammatory reaction that can be devastating. We report the condition in a nine-year-old boy who presented with mild right upper lid swelling wherein cysticercus cellulosae cyst was found within the mass of the right lateral rectus muscle. It becomes important to report this case because of the unusual site of the cyst which masqueraded as a lipo-dermoid.

Neurophysiological features of tactile versus visual guidance of ongoing movement.

Although studies have investigated tactile and visual processing for perception, sensory processing for ongoing action remains poorly understood. The purpose of this study was to explore modality-specific patterns of cortical activation and functional connectivity in a practiced trajectory tracking task. Participants traced irregular shapes with their index finger using either touch or vision for guidance. In 60 tactile-motor (TM) trials, movement was guided only by tactile feedback of semicircular bumps on a plastic tile. In 60 visuo-motor (VM) trials, movement was guided only by vision of dots on a screen seen through a small window at the finger tip. The order of TM and VM trials was counterbalanced across 16 participants. Electroencephalography (EEG) was used to estimate cortical activation (task-related spectral power) and functional connectivity (task-related magnitude-squared coherence) in the alpha (8-12 Hz) and beta (13-30 Hz) bands during the last 12 movement trials in each sensorimotor task. TM vs. VM tasks exhibited a larger drop in global beta cortical activation, and greater alpha coherence between central, parietal, and occipital regions. VM vs. TM tasks were characterized by low global alpha coherence. Trace time and cortical activation of the last 12 VM trials were reduced in the group performing the VM task after the TM task compared to those performing the VM task first. Beta connectivity initiated by the first task was maintained on the subsequent second task, regardless of the task order. Identification of modality- and order-specific EEG characteristics provides insight into the sensory control of movement.

Hearing Safety From Single- and Double-Pulse Transcranial Magnetic Stimulation in Children and Young Adults.

PURPOSE: Concerns regarding hearing safety have limited the number of studies using transcranial magnetic stimulation (TMS) in children and young adults. The objective of this study was to examine the safety of TMS with regards to hearing in a group of 16 children and young adults (17.3 ± 4.9 years) with and without brain injury. METHODS: Pure-tone hearing thresholds and distortion-product otoacoustic emissions were measured before and after exposure to single- and paired-pulse TMS (1-2 sessions of 149-446 TMS pulses at a median of 49%-100% maximum stimulator output over a 2.2 hours period). RESULTS: No mean change in hearing outcomes was noted. In addition, no clinically significant change in hearing threshold was observed in any participant, and participants did not experience a subjective change in hearing after TMS exposure. CONCLUSIONS: Single- and double-pulse TMS administered within the parameters used in this study, which included hearing protection, can be used in children and young adults without impacting hearing. This study provides further evidence for hearing safety after TMS exposure in children and young adults.

Inter-joint coordination analysis of reach-to-grasp kinematics in children and adolescents with obstetrical brachial plexus palsy.

BACKGROUND: Obstetrical brachial plexus palsy is a common birth injury to nerves passing through the brachial plexus that may result in structural and functional abnormalities. Individual joint trajectories from kinematic analyses have been used to evaluate the source and extent of abnormalities. Here, two summary measures of limb kinematics were utilized: 1) the Arm Profile Score summarizing upper limb joint kinematic abnormalities from a typical pattern across a task, and 2) the recently developed Multi-joint Coordination Measure using principal component analysis to characterize typical coordination of multiple joints throughout a task and compute deviations in time and space. Our aim was to compare these kinematic measures in persons with and without injury and relate these to clinical and functional scales. METHODS: 3D kinematic data from 10 upper limb joints were collected on 15 children and adolescents with obstetrical brachial plexus palsy and 21 controls during a reach-to-grasp task in both limbs. The two kinematic measures were computed and correlated with each other and the Mallet and ABILIHAND-Kids. FINDINGS: Both measures revealed that joint angles primarily contributing to shoulder and wrist motion were most prominently affected in the non-dominant limb in obstetrical brachial plexus palsy, with the Multi-joint Coordination Measure additionally indicating when in the motion coordination worsens. These were moderately interrelated but neither correlated with other scales. INTERPRETATION: The Multi-joint Coordination Measure, while related to the Arm Profile Score, may have additional utility for individualized treatment planning and evaluation of any motor task due to the unique spatial-temporal information provided.

Compound composite odontoma.

The term odontoma has been used as a descriptor for any tumor of odontogenic origin. It is a growth in which both epithelial and mesenchymal cells exhibits complete differentiation. Odontomas are considered as hamartomas rather than true neoplasm. They are usually discovered on routine radiographic examination. Odontomas, according to the World Health Organization, are classified into complex odontoma and compound odontomas. The present paper reports a case of compound composite odontomas.

Probing the interaction of the ipsilateral posterior parietal cortex with the premotor cortex using a novel transcranial magnetic stimulation technique.

OBJECTIVE: Functional imaging studies have shown that control of planned movement involves a distributed network that involves the premotor (PMv) and posterior parietal cortices (PPC). Similarly, anatomical studies show that these regions are densely interconnected via white matter tracts. We therefore hypothesized that the PPC influence over the motor cortex is partly via a connection with the PMv. METHODS: Using a novel three-pulse ipsilateral transcranial magnetic stimulation technique, we preconditioned the PPC (80% RMT) at ISIs from 4-15 ms prior to stimulating the PMv and M1 at ISIs of 4 and 6 ms. RESULTS: As previously shown, PMv-M1 paired-pulse stimulation resulted in inhibition of the MEP (90% RMT, 4-6 ms) and PPC-M1 paired-pulse stimulation resulted in facilitation of the MEP (90% RMT, 4-8 ms). PPC-M1 paired-pulse stimulation at 80% RMT preconditioning had no effect on M1. PPC-PMv-M1 stimulation resulted in reversal of inhibition observed with PMv-M1 stimulation at ISIs ranging from 6 to 15 ms. CONCLUSIONS: The reversal of inhibition observed with PPC-PMv-M1 stimulation suggests that the parietal connection to the PMv plays a role in the modulation of M1. SIGNIFICANCE: This is the first study to stimulate three intrahemispheric regions in order to test a disynaptic connection with M1. The described network may be important in a variety of movement disorders.

Coordination of Reach-to-Grasp Kinematics in Individuals With Childhood-Onset Dystonia Due to Hemiplegic Cerebral Palsy.

Functional reaching is impaired in dystonia. Here, we analyze upper extremity kinematics to quantify timing and coordination abnormalities during unimanual reach-to-grasp movements in individuals with childhood-onset unilateral wrist dystonia. Kinematics were measured during movements of both upper limbs in a patient group ( n = 11, age = 17.5 ±5 years), and a typically developing control group ( n = 9, age = 16.6 ±5 years). Hand aperture was computed to study the coordination of reach and grasp. Time-varying joint synergies within one upper limb were calculated using a novel technique based on principal component analysis to study intra-limb coordination. In the non-dominant arm, results indicate reduced coordination between reach and grasp in patients who could not lift the grasped object compared to those who could lift it. Lifters exhibit incoordination in distal upper extremity joints later in the movement and non-lifters lacked coordination throughout the movement and in the whole upper limb. The amount of atypical coordination correlates with dystonia severity in patients. Reduced coordination during movement may reflect deficits in the execution of simultaneous movements, motor planning, or muscle activation. Rehabilitation efforts can focus on particular time points when kinematic patterns deviate abnormally to improve functional reaching in individuals with childhood-onset dystonia.

Distinct interneuronal networks influence excitability of the surround during movement initiation.

Surround inhibition (SI) is a feature of motor control in which activation of task-related muscles is associated with inhibition of neighboring, nonprotagonist muscles, allowing selective motor control. The physiological basis for SI still remains unknown. In all previous studies, SI in the motor system was measured during movement initiation by using transcranial magnetic stimulation (TMS) to deliver a posteroanterior current at a single suprathreshold intensity. To expand our understanding of SI, we explored this phenomenon at a wide range of intensities and by stimulating motor cortex with currents along anteroposterior and lateromedial directions. Fifteen healthy volunteers performed a brief isometric index finger flexion on hearing a tone. Electromyography was recorded from the synergist and surround finger muscles. Single-pulse TMS was applied to stimulate the surround muscle at different intensities at rest or movement initiation. The motor evoked potential (MEP) amplitudes were then plotted against stimulation intensities to obtain the MEP recruitment curves for the rest and movement initiation conditions and for the three current directions for every subject. From the recruitment curves, we found that surround inhibition could be elicited only by the posteroanterior current. Hence, we postulate that surround inhibition is mediated by intracortical circuits in the motor cortex. Also, for the first time, we observed surround facilitation when the motor cortex was stimulated with anteroposterior current. Further studies are needed to investigate the mechanisms underlying both these phenomena individually in healthy subjects and patients with dystonia and other movement disorders.

Cortical activation and inter-hemispheric sensorimotor coherence in individuals with arm dystonia due to childhood stroke.

OBJECTIVE: Dystonia is a disabling motor disorder often without effective therapies. To better understand the genesis of dystonia after childhood stroke, we analyzed electroencephalographic (EEG) recordings in this population. METHODS: Resting spectral power of EEG signals over bilateral sensorimotor cortices (Powrest), resting inter-hemispheric sensorimotor coherence (Cohrest), and task-related changes in power (TRPow) and coherence (TRCoh) during wrist extension were analyzed in individuals with dystonia (age 20±3years) and healthy volunteers (age 17±5years). RESULTS: Ipsilesional TRPow decrease was significantly lower in patients than controls during the more affected wrist task. Force deficits of the affected wrist correlated with reduced alpha TRPow decrease on the ipsilesional and not the contralesional hemisphere. Cohrest was significantly lower in patients than controls, and correlated with more severe dystonia and poorer hand function. Powrest and TRCoh were similar between groups. CONCLUSIONS: The association between weakness and cortical activation during wrist extension highlights the importance of ipsilesional sensorimotor activation on function. Reduction of Cohrest in patients reflects a loss of inter-hemispheric connectivity that may result from structural changes and neuroplasticity, potentially contributing to the development of dystonia. SIGNIFICANCE: Cortical and motor dysfunction are correlated in patients with childhood stroke and may in part explain the genesis of dystonia.