PELP1 signaling contributes to medulloblastoma progression by regulating the NF-κB pathway

Medulloblastoma (MB) is the most common and deadliest brain tumor in children. Proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) is a scaffolding protein and its oncogenic signaling is implicated in the progression of several cancers. However, the role of PELP1 in the progression of MB remains unknown. The objective of this study is to examine the role of PELP1 in the progression of MB. Immunohistochemical analysis of MB tissue microarrays revealed that PELP1 is overexpressed in the MB specimens compared to normal brain. Knockdown of PELP1 reduced cell proliferation, cell survival, and cell invasion of MB cell lines. The RNA-sequencing analysis revealed that PELP1 knockdown significantly downregulated the pathways related to inflammation and extracellular matrix. Gene set enrichment analysis confirmed that the PELP1-regulated genes were negatively correlated with nuclear factor-κB (NF-κB), extracellular matrix, and angiogenesis gene sets. Interestingly, PELP1 knockdown reduced the expression of NF-κB target genes, NF-κB reporter activity, and inhibited the nuclear translocation of p65. Importantly, the knockdown of PELP1 significantly reduced in vivo MB progression in orthotopic models and improved the overall mice survival. Collectively, these results suggest that PELP1 could be a novel target for therapeutic intervention in MB.     

Respiratory and vasomotor effects of excitatory amino acid on ventral medullary surface

Three glutamic acid analogues, N-methyl-D-aspartic (NMDA), quisqualic (QQ), and kainic (KAI) acids were applied topically to the ventral surface of the medulla (VMS) in paralyzed, vagotomized and carotid sinus denervated cats hyperventilated to apnea. Respiratory and vasomotor effects were assessed by changes in phrenic nerve activity and systemic arterial blood pressure. All three agents to varying degrees raised systemic blood pressure, but only NMDA consistently initiated phrenic nerve activity at pCO2 levels below that observed in control trials. KAI and QQ raised blood pressure even in those animals in which they had little effect on initiating phrenic nerve activity. Furthermore, respiratory responses were obtained from localized areas on VMS, namely the intermedio-caudal zone (I-C areas); whereas blood pressure elevations could be obtained from wider VMS areas including the rostral zone (R areas). In addition, the effects of the three amino acids on blood pressure were quantitatively different with KAI causing much greater increases in blood pressure than QQ or NMDA. The respiratory and vasomotor effects of NMDA and QQ were blocked by the use of 2-amino-5-phosphonovaleric acid and L-glutamic acid diethylester, their respective antagonists. The results suggest that neurons in the VMS which cause respiratory and vasomotor responses are not identical. Cells containing receptors stimulated by NMDA predominantly increase respiration, whereas cells containing receptors excited by KAI are more effective in eliciting vasomotor responses.     

Dynamic Stiffness Matrix Approach to Free Vibration Analysis of Functionally Graded Rotor Bearing System Subjected to Thermal Gradients

The dynamic stiffness matrix (DSM) method, an analytical method that provides exact solutions, has been used for the first time for the free vibration analysis of a functionally graded (FG) rotor bearing system subjected to temperature gradients and to investigate its application to FG rotors. The material gradation occurs based on the power law between the inner metal core and the outer ceramic rich layer of the FG rotor. The temperature gradation follows the Fourier law of heat conduction which leads to non-linear temperature distribution (NLTD) in the radial direction of the FG rotor. The development of the DSM formulations for Timoshenko FG rotor elements using the governing equations derived from translational and rotational equilibrium conditions is the novelty of the present work. The DSM of the FG rotor elements, rigid disk and linear isotropic bearings are assembled to obtain the global dynamic stiffness matrix of the FG rotor bearing system. The natural whirl frequencies are computed from the global DSM using the Wittrick–William algorithm as a root searching technique. The natural and whirl frequencies are validated with the results available in the literature and the exactness of the DSM method has been exemplified.     

Dental crowding in primary dentition and its relationship to arch and crown dimensions among preschool children of Davangere

The purpose of the present study was to evaluate dental crowding in the deciduous dentition and its relationship to the crown and the arch dimensions among preschool children of Davangere. Stratified randomized selection of one hundred, 3-4 year old healthy children with all primary teeth erupted was done and divided into two groups. One group had children with anterior crowding in both the arches while the other had spacing. Alginate impressions of the upper and lower arches were made and the study casts were obtained. The tooth and arch dimensions were determined. Mesiodistal dimensions of all the teeth were significantly larger in the crowded arch group. However, the buccolingual dimensions of the maxillary right central incisor, mandibular lateral incisors and the maxillary molars and the crown shape ratio of maxillary lateral incisors, mandibular canines and mandibular right second molar was statistically different. No significant correlation was found between the arch width and the presence of crowding of deciduous dentition. The arch depth of the spaced dentition was greater when compared to the crowded ones. The arch perimeter of the crowded arches was significantly less than the spaced arches.     

Acute symptomatic seizures due to single CT lesions: how long to treat with antiepileptic drugs?

The duration of anti epileptic drug therapy for single small enhancing CT lesions (SSECTL) presents a major dilemma. We studied the efficacy of short duration (6 months) antiepileptic drug therapy as compared to long duration (2 years) drug therapy. Seventy three patients presenting with seizures and showing SSECTL on cranial CT scans (plain and contrast) were randomized into group A (6 months therapy) and group B (2 years therapy). There were 47 patients in group A and 26 patients in group B. Patients were followed up for one year after withdrawal of anti epileptic drugs. CT Head (plain and contrast) was repeated after 3 months, or earlier in cases of recurrence to rule out reinfection. 53.2% in group A and 53.8% in group B showed complete resolution and were seizure free on one year follow up. Punctate residual calcification was seen in 46.8% in group A and 46.2% in group B. Eight patients (17%) in group A and three (11.5%) in group B had a recurrence. The difference in recurrence of seizure between the two groups was not statistically significant (p<0.77) in the calcified lesion subset. Since none of the patients in total resolution subset showed recurrence, the difference between calcified and total resolution subset was highly significant. The study shows that a short duration (6 months) AED therapy in patients with total resolution of lesion on follow up scan, may be adequate in comparison to those who have calcific speck as a residue. However, a longer duration of therapy in case of calcific group probably does not alter their chances of recurrence.     

Multisite Study of Cryptosporidiosis in Children with Diarrhea in India

Cryptosporidium spp., a common cause of diarrhea in children, were investigated in the first multisite study in India. Diarrheal stools from hospitalized children aged <5 years from Delhi, Trichy, and Vellore were analyzed by microscopy, PCR-restriction fragment length polymorphism (RFLP), and/or sequencing at the small-subunit (SSU) rRNA and Cpgp40/15 loci for species determination and subgenotyping, respectively. Seventy of 2,579 (2.7%) children, 75% of whom were <2 years old, had cryptosporidial diarrhea as determined by microscopy. Genotyping and subgenotyping showed that Cryptosporidium hominis was the most commonly identified species (59/67 children), and subgenotypes Ie, Ia, Ib, and Id were common in all centers. A novel C. parvum subgenotype, IIn, was identified in Vellore. Meteorological analysis revealed a higher rate of cryptosporidial positivity during hotter and drier weather in Delhi.Cryptosporidium spp. are an important cause of endemic parasitic diarrhea in children in developing countries. In addition to causing symptoms associated with watery diarrhea, vomiting, and weight loss, early childhood cryptosporidiosis has been shown by studies to be associated with subsequent faltering of growth (reviewed in reference 11). Cryptosporidium hominis and C. parvum cause the majority of infections in children in developing countries, with C. hominis predominating and occasional reports of infection with zoonotic species such as C. felis, C. canis, C. meleagridis, and C. muris (30). C. hominis infection has been found to be associated with greater levels of oocyst shedding (4) and longer durations of oocyst shedding (31) and diarrhea (15) than C. parvum infection. In a recent community-based study in Vellore, we found increased levels of severity of diarrhea in C. hominis-infected children compared to the levels observed in children infected with other species (1).Cryptosporidium spp. have been classified into several distinct subgenotypes based on extensive polymorphisms in the Cpgp40/15 (also referred to as GP60) locus by use of PCR-restriction fragment length polymorphism (RFLP) or sequencing of PCR products (reviewed in reference 30).A number of studies from India have reported Cryptosporidium spp. in diarrheal stool samples from children, with positivity rates of up to nearly 20% (17) and asymptomatic infection rates of up to 10% (19), using stool microscopy for detection. However, only three studies have used molecular techniques for identification of cryptosporidiosis in children in India (9, 13, 22), suggesting that the actual infection rates may be significantly higher. In a previous hospital-based study in Vellore, we that found that PCR (15.2%) identified more than 3 times the number of cases of cryptosporidial diarrhea than microscopy (4.4%) (2). The aim of the present study was to identify the Cryptosporidium species and Cpgp40/15 subgenotypes associated with cryptosporidial diarrhea in hospitalized children from 3 centers in the country, since no studies have examined cryptosporidiosis using the same methods in more than one location.     

Mapping cis-regulatory domains in the human genome using multi-species conservation of synteny

Our inability to associate distant regulatory elements with the genes they regulate has largely precluded their examination for sequence alterations contributing to human disease. One major obstacle is the large genomic space surrounding targeted genes in which such elements could potentially reside. In order to delineate gene regulatory boundaries, we used whole-genome human-mouse-chicken (HMC) and human-mouse-frog (HMF) multiple alignments to compile conserved blocks of synteny (CBSs), under the hypothesis that these blocks have been kept intact throughout evolution at least in part by the requirement of regulatory elements to stay linked to the genes they regulate. A total of 2116 and 1942 CBSs >200 kb were assembled for HMC and HMF, respectively, encompassing 1.53 and 0.86 Gb of human sequence. To support the existence of complex long-range regulatory domains within these CBSs, we analyzed the prevalence and distribution of chromosomal aberrations leading to position effects (disruption of a gene's regulatory environment), observing a clear bias not only for mapping onto CBS but also for longer CBS size. Our results provide an extensive data set characterizing the regulatory domains of genes and the conserved regulatory elements within them.     

Cerebrospinal fluid profile of amyloid β42 (Aβ42), hTau and ubiquitin in North Indian Alzheimer's disease patients

Alzheimer's disease (AD) is the most common form of dementia, and is characterized by the degeneration of neurons and their synapses, and a higher number of amyloid plaques and neurofibrillary tangles (NFTs) compared with that found in non-demented individuals. Amyloid-β-peptides (Aβ) are major components of amyloid plaques in AD brain whereas NFTs are composed of Tau and associated with ubiquitin. The aim of the present study was to analyze the levels of Aβ42, hTau (total Tau) and ubiquitin in CSF of North Indian population. CSF Aβ42, Tau and ubiquitin were measured in CSF of AD patients as well as controls using ELISA assays. Here we report low Aβ42 levels in AD patients (324.24 ± 76.38 pg/ml) as compared to those in non-AD (NAD) (668.34 ± 43.13 pg/ml), neurological controls (NCs) (727.28 ± 46.49 pg/ml) and healthy controls (HCs) (976.47 ± 124.46 pg/ml). In contrast, hTau and ubiquitin levels were significantly high (568.65 ± 48.89 pg/ml and 36.82 ± 4.34 ng/ml, respectively) in AD patients compared to those in NAD, NC and HC. The hTau levels were 267.37 ± 36.64 pg/ml, 167.34 ± 44.27 pg/ml and 107.62 ± 24.27 pg/ml in NAD, NC and HC, respectively. Similarly, ubiquitin levels were 23.57 ± 2.32 ng/ml, 19.76 ± 3.64 ng/ml and 13.24 ± 4.56 ng/ml in NAD, NC and HC, respectively. In conclusion, low Aβ42 and high Tau–ubiquitin levels were found in North Indian AD patients.