Evaluation of the satisfaction level of patients visiting dental emergency services.

Context: Inadequate data is available of patient satisfaction in dental emergency departments in India. Aim: This study was undertaken with the aim to evaluate the satisfaction level of patients visiting emergency services of a dental institute in an Indian city. Settings and Design: A questionnaire-based cross-sectional exploratory study was designed over a period of 2 months. Subjects and Methods: A total of 51 subjects visiting the dental emergency services after routine working hours participated in this questionnaire-based study and submitted their responses. Statistical Analysis Used: Pearson's Chi-square test. Results: A statistically significant correlation (P < 0.05) was observed between effectiveness of the treatment given in terms of relief from complaints with the experience at reception, rating the hospital in terms of overall waiting time for any service with ambience (P = 0.031), between effectiveness of the treatment given in terms of control/relief from complaints (P = 0.00), 'rating patient's experience with "on-duty doctor" (response time, behavior, appearance, attitude etc.), rating the hospital in terms of overall waiting time for any service (P = 0.010), experience' with nursing staff (responsive, courteous, polite) and rating the hospital in terms of overall waiting time for any service. Conclusions: Emergency care where patients were satisfied included reception, greeting while entering the department, ambience of the hospital and the Emergency Department, and good experience with the on-duty doctor, nursing staff, and security. Waiting time for treatment at emergency care was less. Recommending this hospital to others was statistically significant with the experience of the patient with the staff.

Role and relevance of fish cell lines in advanced in vitro research.

INTRODUCTION: Cell line derived from fish has been established as a promising tool for studying many key issues of aquaculture covering fish growth, disease, reproduction, genetics, and biotechnology. In addition, fish cell lines are very useful in vitro models for toxicological, pathological, and immunological studies. The easier maintenance of fish cell lines in flexible temperature regimes and hypoxic conditions make them preferable in vitro tools over mammalian cell lines. Great excitement has been observed in establishing and characterizing new fish cell lines representing diverse fish species and tissue types. The well-characterized and authenticated cell lines are of utmost essential as these represent cellular functions very similar to in vivo state of an organism otherwise it would affect the reproducibility of scientific research. CONCLUSION: The fish cell lines have exhibited encouraging results in several key aspects of in vitro research in aquaculture including virology, nutrition and metabolism, production of vaccines, and transgenic fish production. The review paper reports the cell lines developed from fish, their characterization, and biobanking along with their potential applications and challenges in in vitro research.

Frequency Dependent Charge Transport and Spin State Switching Characteristics of Fe(phen)2(NCS)2 in Polymer.

We report on the bistability in spin states of spin crossover (SCO) compound Fe(phen)2(NCS)2 in polymer (polypyrrole) by frequency (1-100 kHz) and temperature dependent (305-457 K) electrical conductivity measurements. The structure and growth of SCO compounds in conducting polymer are obtained by scanning electron microscopy, X-ray diffraction and optical absorption measurements. The thermal dependence of ac conductivity σ(ω) shows the clear formation of a hysteresis loop in its cooling and heating cycle due to the difference in conductivity in high spin and low spin state. The size, shape and width of the hysteresis loops are found to be critically dependent on the applied frequency and/or the ratio between SCO and polymer. The ac conductivity is found to exhibit a dispersive behavior following Jonscher's law: σ(ω) ∝ ωn below a critical frequency ωc, above which it is found to monotonically decrease with increasing frequency. The thermal dependence of the exponent n and ωc is also explored. The charge transport phenomena are explained in the framework of hopping of charge carriers. The data reveals that addition of polymer can play an important role to tune the conductivity of SCO compounds and its spin state dependence characteristics which may be quite helpful for fabricating future spin-based devices. Temperature dependent magnetic susceptibility measurement also confirms the spin transition behavior of the SCO/ppy composite samples. These SCO/ppy composite samples can be taken as the reliable nanomaterials fabricated with the concept of future spin based nanoarchitectonics.

Notch and Cdk5 in Zebrafish Mindbomb Mutant: Co-regulation or Coincidence?

Notch signalling is critical for the development of the nervous system. In the zebrafish mindbomb mutants, disruption of E3 ubiquitin ligase activity inhibits Notch signalling. In these mutant embryos, precocious development of primary neurons leading to depletion of neural progenitor cells results in a neurogenic phenotype characterized by defects in neural patterning and brain development. Cyclin-dependent kinase 5 (Cdk5), a predominant neuronal kinase, is involved in a variety of essential functions of the nervous system. Most recently, mammalian studies on Notch and Cdk5 regulating each other's function have been emerging. The status of Cdk5 in the mindbomb mutant embryos with excessive primary neurons is not known. In situ hybridization of the zebrafish mindbomb mutant embryos uncovered a robust upregulation in Cdk5 expression but with a reduced Cdk5 activity. The implications of these findings in both the mammalian system and zebrafish are discussed in this mini-review to provide a glimpse into the relationship between Notch and Cdk5 that may explain certain neurodevelopmental defects associated with either mutations in ubiquitin ligase or altered expression of Cdk5.

Development of Escherichia coli-based gene expression profiling of sewage sludge leachates.

AIMS: The impact of municipal waste on pathogenic micro-organisms released into the environment is a public health concern. This study aims to evaluate the effects of sewage sludge and antibiotic contaminants on stress response, virulence and antibiotic resistance in a pathogenic Escherichia coli. METHODS AND RESULTS: The effects of sewage sludge leachates on uropathogenic E. coli CFT073 were determined by monitoring the expression of 45 genes associated with antibiotic/metal resistance, stress response and virulence using RT-qPCR. The E. coli gene expression was validated using subinhibitory concentrations of tetracycline and ciprofloxacin. E. coli exposed to sewage sludge or sewage sludge+fly ash leachates altered the expression of five antibiotic and metal resistance, three stress response and two virulence-associated genes. When antibiotics were combined with sludge or sludge+fly ash the antibiotic-associated gene expression was altered. CONCLUSIONS: E. coli treated with two sludge leachates had distinct gene expression patterns that were altered when the sludge leachates were combined with tetracycline, although to a lesser extent with ciprofloxacin. SIGNIFICANCE AND IMPACT OF THE STUDY: The E. coli multigene expression analysis is a potential new tool for assessing the effects of pollutants on pathogenic microbes in environmental waters for improved risk assessment.

Dynamics of microvesicle generation in B-cell chronic lymphocytic leukemia: implication in disease progression.

Previously, we reported that B-cell chronic lymphocytic leukemia (CLL) patients contained elevated levels of microvesicles (MVs). However, given the quiescent nature of CLL B-cells and the relative indolence of the disease, the dynamics of MV generation and their unique phenotypes are not clearly defined. In this study, we find that CLL B-cells generate MVs spontaneously and can be further induced by B-cell receptor-ligation. Most interestingly, CLL B-cells predominantly generate CD52+ MVs, but not CD19+ MVs in vitro, suggesting preferential usage of CD52 into leukemic-MVs and that the CLL plasma MV phenotypes corroborate well with the in vitro findings. Importantly, we detected increased accumulation of CD52+ MVs in previously untreated CLL patients with progressive disease. Finally, sequential studies on MVs in pre- and post-therapy CLL patients demonstrate that although the plasma CD52+ MV levels drop significantly after therapy in most and remain at low levels in some patients, a trend of increased accumulation of CD52+ MVs was detected in majority of post-therapy CLL patients (25 of 33). In total, this study emphasizes that dynamic accumulation of CD52+ MVs in plasma can be used to study CLL progression and may be a useful biomarker for patients as they progress and require therapy.

MicroRNA-101 regulated transcriptional modulator SUB1 plays a role in prostate cancer.

MicroRNA-101, a tumor suppressor microRNA (miR), is often downregulated in cancer and is known to target multiple oncogenes. Some of the genes that are negatively regulated by miR-101 expression include histone methyltransferase EZH2 (enhancer of zeste homolog 2), COX2 (cyclooxygenase-2), POMP (proteasome maturation protein), CERS6, STMN1, MCL-1 and ROCK2, among others. In the present study, we show that miR-101 targets transcriptional coactivator SUB1 homolog (Saccharomyces cerevisiae)/PC4 (positive cofactor 4) and regulates its expression. SUB1 is known to have diverse role in vital cell processes such as DNA replication, repair and heterochromatinization. SUB1 is known to modulate transcription and acts as a mediator between the upstream activators and general transcription machinery. Expression profiling in several cancers revealed SUB1 overexpression, suggesting a potential role in tumorigenesis. However, detailed regulation and function of SUB1 has not been elucidated. In this study, we show elevated expression of SUB1 in aggressive prostate cancer. Knockdown of SUB1 in prostate cancer cells resulted in reduced cell proliferation, invasion and migration in vitro, and tumor growth and metastasis in vivo. Gene expression analyses coupled with chromatin immunoprecipitation revealed that SUB1 binds to the promoter regions of several oncogenes such as PLK1 (Polo-like kinase 1), C-MYC, serine-threonine kinase BUB1B and regulates their expression. Additionally, we observed SUB1 downregulated CDKN1B expression. PLK1 knockdown or use of PLK1 inhibitor can mitigate oncogenic function of SUB1 in benign prostate cancer cells. Thus, our study suggests that miR-101 loss results in increased SUB1 expression and subsequent activation of known oncogenes driving prostate cancer progression and metastasis. This study therefore demonstrates functional role of SUB1 in prostate cancer, and identifies its regulation and potential downstream therapeutic targets of SUB1 in prostate cancer.

Regulation of complement-dependent cytotoxicity by TGF-ß-induced epithelial-mesenchymal transition.

The process of epithelial-mesenchymal transition (EMT), in addition to being an initiating event for tumor metastasis, is implicated in conferring several clinically relevant properties to disseminating cancer cells. These include stem cell-like properties, resistance to targeted therapies and ability to evade immune surveillance. Enrichment analysis of gene expression changes during transforming growth factor-ß (TGF-ß)-induced EMT in lung cancer cells identified complement cascade as one of the significantly enriched pathway. Further analysis of the genes in the complement pathway revealed an increase in the expression of complement inhibitors and a decrease in the expression of proteins essential for complement activity. In this study, we tested whether EMT confers resistance to complement-dependent cytotoxicity (CDC) in lung cancer cells and promotes tumor progression. CD59 is a potent inhibitor of membrane attack complex that mediates complement-dependent cell lysis. We observed a significant increase in the CD59 expression on the surface of cells after TGF-ß-induced EMT. Furthermore, CD59 knockdown restored susceptibility of cells undergoing EMT to cetuximab-mediated CDC. TGF-ß-induced CD59 expression during EMT is dependent on Smad3 but not on Smad2. Chromatin immunoprecipitation analysis confirmed that Smad3 directly binds to the CD59 promoter. Stable knockdown of CD59 in A549 cells inhibited experimental metastasis. These results demonstrate that TGF-ß-induced EMT and CD59 expression confers an immune-evasive mechanism to disseminating tumor cells facilitating tumor progression. Together, our data demonstrates that CD59 inhibition may serve as an adjuvant to enhance the efficacy of antibody-mediated therapies, as well as to inhibit metastasis in lung cancer.

A multigene array for measurable residual disease detection in AML patients undergoing SCT.

AML is a diagnosis encompassing a diverse group of myeloid malignancies. Heterogeneous genetic etiology, together with the potential for oligoclonality within the individual patient, have made the identification of a single high-sensitivity marker of disease burden challenging. We developed a multiple gene measurable residual disease (MG-MRD) RQ-PCR array for the high-sensitivity detection of AML, retrospectively tested on 74 patients who underwent allo-SCT at the NHLBI in the period 1994-2012. MG-MRD testing on peripheral blood samples prior to transplantation demonstrated excellent concordance with traditional BM-based evaluation and improved risk stratification for post-transplant relapse and OS outcomes. Pre-SCT assessment by MG-MRD predicted all clinical relapses occurring in the first 100 days after allo-SCT compared with 57% sensitivity using WT1 RQ-PCR alone. Nine patients who were negative for WT1 prior to transplantation were correctly reclassified into a high-risk MG-MRD-positive group, associated with 100% post-transplant mortality. This study provides proof of principle that a multiple gene approach may be superior to the use of WT1 expression alone for AML residual disease detection.

Evaluation of dental implant insertion torque using a manual ratchet.

BACKGROUND: Dental implant insertion torque is crucial for the success of the implant and the prosthesis. This in-vivo study was undertaken to determine the average insertion torque being applied to the dental implant while surgically placing it with a non-calibrated manual ratchet. METHODS: Three dental surgeons placed a total of 45 dental implants (Touareg, ADIN, Afula, Israel) in 42 selected patients. Each surgeon placed 15 implants. Standardised protocols were followed to prepare the site to place the dental implant. Each implant was placed using a manual non-calibrated implant ratchet first. Once the implant was nearly placed, a manual calibrated torque gauge ratchet was used to place the implant in its final position and at that instance, the maximum final torque applied was noted on the torque gauge scale. RESULTS: The mean dental implant insertion torque applied by three surgeons using a non-calibrated manual ratchet was estimated to be 63.26 Ncm with a standard error of 6.80 i.e. (63.26 + 6.8), which was significantly higher than the baseline of 35 Ncm (p < 0.0001). The mean dental implant torque applied by Surgeon 1, 2 and 3, respectively, was 65.93 Ncm, 62.60 Ncm and 62.13 Ncm and this difference amongst them was found to be statistically insignificant (p > 0.05) and each of them had reached more than the baseline level of 35 Ncm individually and significantly (p < 0.0001). CONCLUSION: Without the use of torque measuring devices, an average surgeon may achieve an average insertion torque of 63.26 + 6.8 Ncm.

Establishment and characterization of a piscean PCF cell line for toxicity and gene expression studies as in vitro model.

A new piscean fibroblastic cell line termed as PCF derived from the caudal fin tissue of dark mahseer, Puntius (Tor) chelynoides was established and characterized in the present study which was found to be suitable for toxicity and gene expression studies as in vitro model. The cell line grew well in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum (FBS). The cells were able to grow at a temperature ranging from 20 to 28 °C with an optimal growth at 24 °C and the cell line have been expanded in culture for more than 70 passages. Authentication of the cell line was carried out using mitochondrial DNA markers (Cytochrome Oxidase subunit I and 16S ribosomal RNA). Presence of vimentin in the cells confirmed the fibroblastic origin of cell line. Significant cytopathic effects were observed upon exposure of PCF cell line to bacterial extracellular products and the study also validated the suitability of cell line in transgenic applications as well as in genotoxicity assessment as an in vitro model.

Evaluation of shear bond strength of composite resin bonded to alloy treated with sandblasting and electrolytic etching.

Conservation of natural tooth structure precipitated the emergence of resin-retained fixed partial dentures. The weakest link in this modality is the bond between resin cement and alloy of the retainer. Various alloy surface treatment have been recommended to improve alloy-resin bond. This in vitro study was carried out to observe changes in the Nickel-Chromium alloy (Wiron 99, Bego) surface following sandblasting or electrolytic etching treatment by scanning electron microscope (SEM) and to evaluate the shear bond strength of a resin luting cement bonded to the surface treated alloy. 80 alloy blocks were cast and divided into four groups of 20 each. In groups-A & B, the test surfaces were treated by sandblasting with 50 and 250 µm sized aluminium oxide particles respectively. In groups-C & D, the test surfaces were first treated by sandblasting with 50 and 250 µm sized aluminium oxide particles respectively followed by electrolytic etching. Test surfaces were observed under SEM at 1,000× magnification. Two alloy blocks of each group were luted together by a resin luting cement (Rely X, 3M) and their shear bond strength was tested. The mean shear bond strength in MPa of groups-A to D were 6.44 (±0.74), 8.18 (±0.51), 14.45 (±0.59) and 17.43 (±1.20) respectively. Group-D showed bond strength that is more than clinically acceptable bond strength. It is recommended that before luting resin-retained fixed partial dentures, the fitting surface of the retainer should be electrolytically etched to achieve adequate micromechanical retention.

Radial glial cell: critical functions and new perspective as a steroid synthetic cell.

The radial glial cell (RGC) is a glial cell type in the central nervous system of all vertebrates. Adult teleost fish have abundant RGCs in the brain in contrast to mammals. Adult fish RGCs have many important functions, including forming a structural scaffold to guide neuronal migration and serving as the progenitor cells in the brain to generate neurons. The role of the RGC in adult neurogenesis explains the high regenerative capacity of adult fish brain. There is increasing evidence from several species that some glial cells produce or metabolize steroids. It is now well-known that teleost RGCs express aromatase and produce estrogens from androgen precursors, which may be important for local neuroendocrine functions and regulation of neurogenesis. The question of whether RGCs are capable of de novo steroid synthesis from cholesterol remains unanswered. However, the expression of steroidogenic acute regulatory protein, and the key enzyme cytochrome P450 17alpha-hydroxylase in primary cultures of goldfish RGCs indicate the potential to produce 17α-hydroxy-pregnenolone and thus other steroid intermediates. The possibility of synthesizing additional non-estrogenic steroids may indicate new functions for the RGC.

Genetic basis of dental agenesis--molecular genetics patterning clinical dentistry.

Tooth agenesis is one of the most common congenital malformations in humans. Hypodontia can either occur as an isolated condition (non-syndromic hypodontia) or can be associated with a syndrome (syndromic hypodontia), highlighting the heterogeneity of the condition. Though much progress has been made to identify the developmental basis of tooth formation, knowledge of the etiological basis of inherited tooth loss is still lacking. To date, the mutation spectra of non-syndromic form of familial and sporadic tooth agenesis in humans have revealed defects in various such genes that encode transcription factors, MSX1 and PAX9 or genes that code for a protein involved in canonical Wnt signaling (AXIN2), and a transmembrane receptor of fibroblast growth factors (FGFR1). The aim of this paper is to review the current literature on the molecular mechanisms responsible for selective hypodontia in humans and to present a detailed overview of causative genes and syndromes associated with hypodontia.

Microwave, infrared-microwave double resonance, and theoretical studies of C2H4···H2S complex.

In this manuscript, rotational spectra of four new isotopologues of the S-H···π bonded C2H4···H2S complex, i.e., C2D4···H2S, C2D4···D2S, C2D4···HDS, and (13)CCH4···H2S have been reported and analyzed. All isotopologues except C2D4···HDS show a four line pattern whereas a doubling of the transition frequencies was observed for C2D4···HDS. These results together with our previous report on the title complex [M. Goswami, P. K. Mandal, D. J. Ramdass, and E. Arunan, Chem. Phys. Lett. 393(1-3), 22-27 (2004)] confirm that both subunits (C2H4 and H2S) are involved in large amplitude motions leading to a splitting of each rotational transition to a quartet. Further, the results also confirm that the motions which are responsible for the observed splittings involve both monomers. Molecular symmetry group analysis, considering the interchange of equivalent H atoms in H2S and C2H4 could explain the observed four line pattern and their intensities in the microwave spectrum. In addition, hydride stretching fundamentals of the complex were measured using coherence-converted population transfer Fourier Transform Microwave-infrared (IR-MW double resonance) experiments in the S-H and C-H stretch regions. Changes in the tunneling splittings upon vibrational excitation are consistent with the isotopic dependence of pure rotational transitions. A complexation shift of 2.7-6.5 cm(-1) has been observed in the two fundamental S-H stretching modes of the H2S monomer in the complex. Vibrational pre-dissociation in the bound S-H stretch has been detected whereas the instrument-limited line-shapes in other S-H and C-H stretches indicate slower pre-dissociation rate. Some local perturbations in the vibrational spectra have been observed. Two combination bands have been observed corresponding to both the S-H stretching fundamentals and what appears to be the intermolecular stretching mode at 55 cm(-1). The tunneling splitting involved in the rotation of C2H4 unit has been deduced to be 1.5 GHz from the IR-MW results. In addition, ab initio barrier heights derived for different motions of the monomers support the experimental results and provide further insight into the motions causing the splitting.

Effect of various remineralising agents on human eroded enamel of primary teeth.

AIM: To determine the effect of three remineralising agents on human primary anterior teeth, i.e. CPP-ACP, CPP-ACPF, fluoridated toothpaste and artificial saliva as control. STUDY DESIGN: Forty primary maxillary anterior teeth were divided into four groups: Group I: CPP-ACP, Group II: CPP-ACPF and Group III: fluoridated toothpaste as experimental and Group IV: artificial saliva as control. The samples were immersed in cola soft drink and artificial saliva for 10 cycles of 5 s each. After erosive procedure, a thin layer of CPP-ACP, CPP-ACPF or fluoridated toothpaste were applied to the tooth surfaces for 3 min, and kept in artificial saliva for 8 h. Samples in the control group were directly kept in artificial saliva without any treatment. The Knoop microhardness of the labial surface of enamel was measured at baseline, after erosion and after the remineralisation procedures. RESULTS: No significant differences were observed among the different groups at pre-erosion and post-erosion intervals. However, after remineralisation, mean microhardness in different groups was significantly higher in Group II as compared to all the other groups (p < 0.001). Group IV had significantly lower mean microhardness as compared to all the other groups (p < 0.001). STATISTICS: The collected data were statistically analysed using ANOVA test followed by Tukey's HSD test as the post-hoc tests to compare the differences in mean microhardness at different time intervals. Paired t test was used to assess the change in mean microhardness within a group. CONCLUSION: CPP-ACPF showed the best remineralisation potential.

Induction of high-titer IgG antibodies against multiple leukemia-associated antigens in CML patients with clinical responses to K562/GVAX immunotherapy.

The ability to target myeloid leukemia with immunotherapy would represent a significant therapeutic advance. We report here immunological analysis of clinical trials of primary and secondary vaccination with K562/GM-CSF immunotherapy in adult chronic phase chronic myeloid leukemia patients (CML-CP) with suboptimal responses to imatinib mesylate. Using serological analysis of recombinant cDNA expression libraries of K562 with autologous vaccinated patient serum, we have identified 12 novel chronic myeloid leukemia-associated antigens (LAAs). We show that clinical responses following K562/GM-CSF vaccination are associated with induction of high-titer antibody responses to multiple LAAs. We observe markedly discordant patterns of baseline and induced antibody responses in these identically vaccinated patients. No single antigen was recognized in all responses to vaccination. We demonstrate that an additional 'booster' vaccination series can be given safely to those with inadequate responses to initial vaccination, and is associated with more frequent induction of IgG responses to antigens overexpressed in K562 vaccine compared with primary CML-CP. Finally, those with induced immune responses to the same LAAs often shared HLA subtypes and patients with clinical responses following vaccination recognized a partially shared but non-identical spectrum of antigens; both findings have potentially significant implications for cancer vaccine immunotherapy.