Genes & Genomics

Comparative gene expression profiling reveals key pathways and genes different in skin epidermal stem cells and corneal epithelial cells Abstract Background Corneal epithelial cells (CECs) are required for corneal transparency and visual function, and corneal injuries may cause corneal blindness. Skin epidermal stem cells (SESCs), which share the same origin with CECs and have the potential of multi-directional differentiation are ideal seed cells for tissue engineered corneal construction to treat corneal blindness. Objective This study aims to investigate critical genes and pathways that may modulate the transdifferentiation from SESCs to CECs. Methods Isolated SESCs and CECs were used for gene expression analysis by microarray. GO and KEGG pathway of differently expressed genes (DEGs) were enriched using DAVID. The protein–protein interaction (PPI) network were then constructed using Cytoscape and highly interconnected module was subsequently isolated from the network by Molecular Complex Detection. Expression of the hub genes and other selected genes were then verified by qRT-PCR. Results We found 112 upregulated and 105 downregulated genes in CECs compared with SESCs. These DEGs were significantly enriched in focal adhesion, PI3K–Akt and TNF signaling pathway. Highly interconnected module of PPI network contains ten genes. Further regulatory network of these genes showed that ESR1 and SLC2A4 were hub genes. Conclusion Our study identified gene expression in SESCs and CECs and suggested that several crucial genes and pathways may play critical roles in transdifferentiation from SESCs to CECs. It may help uncover molecular mechanisms and offer a foundation for promoting tissue-engineered cornea into clinical application.

Karyotype and B chromosome variation in Lilium amabile Palibin Abstract Objective Lilium amabile Palibin (2n = 2x = 24) is an endemic lily species in Korea. B chromosomes are supernumerary chromosomes and the presence of B chromosome in L. amabile was known by previous researches. The current research was conducted to characterize the genetical and cytological features of the B chromosome plants in L. amabile. Methods Karyotype and B chromosome cytotype analyses were carried out among 135 L. amabile accessions that were collected from six geographical locations in Korea using conventional aceto-carmine staining as well as FISH technique with ribosomal RNA gene probes. Results The karyotype of L. amabile genome consisted of two large metacentric, four intermediate subtelocentric, and six intermediate to small acrocentric chromosomes in which chromosomes 1, 6 and 7 carried the 45S rRNA gene loci and chromosome 3 carried the 5S rRNA gene. There were 4 types of B chromosomes, two large B chromosomes and two small B chromosomes. The ribosomal RNA gene loci were not present in the B chromosomes. The 135 accessions were classified into 13 cytotypes including diploids and different B chromosome aneuploids. Among the aneuploids, the most frequent cytotype was 24 + 1B, which was followed by 24 + 2B, 24 + 1b, 24 + 1B + 2b, 24 + 1B + 4b, and 24 + 2B + 4b. Conclusion The karyotype of L. amabile was consistent with other species in the genus Lilium without polyploids. The B chromosome cytotypes were highly variable and the occurrences of different cytotypes were random among the six populations, implying that the B and b chromosome occurrence was random in each population.

Whole RNA-sequencing and gene expression analysis of Trichoderma harzianum Tr-92 under chlamydospore-producing condition Abstract Background Trichoderma is one of the most important biocontrol fungi, which could produce mycelia, conidiospores, and chlamydospores three types of propagules under different conditions. Chlamydospores are produced in harsh conditions in various fungi, and may be more resistant to adverse conditions. However, the knowledge associated with the mechanism of chlamydospore formation remained unclear in Trichoderma. Objectives This study is aimed to explore the essential genes and regulatory pathways associated with chlamydospore formation in Trichoderma. Methods The culture condition, survival rate, and biocontrol effects of chlamydospores and conidiospores from Trichoderma.harzianum Tr-92 were determined. Furthermore, the whole transcriptome profiles of T. harzianum Tr-92 under chlamydospore-producing and chlamydospore-nonproducing conditions were performed. Results T. harzianum Tr-92 produced chlamydospores under particular conditions, and chlamydospore-based formulation of T. harzianum Tr-92 exhibited higher biocontrol ability against Botrytis cinerea in cucumber than conidoiospore-based formulation. In the transcriptome analysis, a total of 2,029 differentially expressed genes (DEGs) were identified in T. harzianum Tr-92 under chlamydospore-producing condition, compared to that under chlamydospore-nonproducing condition. GO classification indicated that the DEGs were significantly enriched in 284 terms among biological process, cellular components and molecular function categories. A total of 19 pathways were observed with DEGs by KEGG analysis. Furthermore, fifteen DEGs were verified by quantitative real-time PCR, and the expression profiles were consistent with the transcriptome data. Conclusion The results would provide a basis on the molecular mechanisms underlying Trichoderma sporulation, which would assist the development and application of fungal biocontrol agents.

Identification and expression analysis of ERECTA family genes in grape ( Vitis vinifera L.) Abstract Background ERECTA family (ERf) genes are found in many dicots and monocots, and play important roles in plant developmental processes and stress responses. However, there is little known on ERf genes in grape (Vitis vinifera L.). Objectives The primary objective of this study was to identify the ERf genes in grape, and to analyze their expression profiles in different organs, during development, and in response to hormone treatments and abiotic/biotic stresses. Methods ERf protein sequences of dicots were aligned in the grape genome (V. vinifera cv. Pinot Noir, PN40024, 12X) with Blast server. The locus tags obtained were inputted in NCBI to retrieve corresponding nucleotide and protein accession numbers. The subcellular localization experiment was performed by the transient expression of VvERECTA-GFP and VvERL2-GFP in mesophyll protoplasts of Arabidopsis. The expression levels of ERf genes in grape leaves were detected by qRT-PCR after hormone treatments and abiotic/biotic stresses. Results We first identified the ERf genes in the grape genome, including VvERECTA and VvERL2. Their cDNA full-length sequences were obtained with the accession numbers MG601756 and MG601757. The result of subcellular localization indicated that the fusion proteins of VvERECTA and VvERL2 were localized in the plasma membrane. There were four conserved domains identified in VvERECTA and VvERL2, including a LRRNT-2, a LRR, a transmembrane and a protein kinase domain. The grape ERf genes expressed highly in young aboveground organs. As grape leaves or berries becoming mature, VvERECTA expressed in a declining trend. The transcript abundance of VvERL2 decreased during leaves development, but showed no significant differences during berries development. The hormone treatments of ABA, SA, MeJA and BR could induce the expression of VvERECTA and VvERL2. The treatments of heat, drought, downy and powdery mildew significantly increased the expression levels of the grape ERf genes. Conclusion The grape ERECTA gene family might play crucial roles in response to abiotic and biotic stresses. We provide the first description of the grape ERf genes and the most comprehensive analysis of their expressions in different biological processes.

In vivo demonstration of enhanced binding between β-clamp and DnaE of pol III bearing consensus i-CBM Abstract Background Among several key protein–protein and protein–DNA interactions within the replisome, the interaction between β-clamp and the DNA polymerase (Pol) III is of crucial importance. This interaction is mediated by a five or six-residue conserved sequence of the DnaE subunit of Pol III, referred to as the Clamp Binding Motif (CBM). In E. coli, DnaE contains two CBMs designated as e-CBM and i-CBM. A consensus sequence (QL[S/D]LF) for the CBMs has previously been proposed and studies involving mutagenesis of both the CBMs have evaluated their protein-binding properties. Surface Plasmon Resonance has been used to show that replacing i-CBM in DnaE with the consensus sequence enhances its binding to β-clamp 120-fold. Objective The current study was aimed to evaluate in vivo interaction between DnaE bearing the consensus i-CBM and β-clamp. Method The C-terminal 405 residues of DnaE, bearing either the consensus i-CBM or the WT i-CBM, with β-clamp were co-expressed in E. coli followed by co-purification of the protein complexes. The interaction was assessed by the ability of the co-expressed proteins to form stable complexes during both affinity and gel filtration chromatography. Result The interaction of β-clamp with DnaEΔ755M containing the consensus i-CBM was found to be more stable than with WT DnaEΔ755, consistent with the in vitro data previously reported. Conclusion The presence of the pieces of sheared DNA generated during sonication promote the interaction of DnaEΔ755M with β-clamp by binding the OB-fold of DnaEΔ755M and β-clamp and serves as a bridge between them.

Construction of genetic linkage map and identification of QTLs related to agronomic traits in DH population of maize ( Zea mays L.) using SSR markers Abstract Background In this study, we used phenotypic and genetic analysis to investigate Double haploid (DH) lines derived from normal corn parents (HF1 and 11S6169). DH technology offers an array of advantages in maize genetics and breeding as follows: first, it significantly shortens the breeding cycle by development of completely homozygous lines in two or three generations; and second, it simplifies logistics, including requiring less time, labor, and financial resources for developing new DH lines compared with the conventional RIL population development process. Objectives In our study, we constructed a maize genetic linkage map using SSR markers and a DH population derived from a cross of normal corn (HF1) and normal corn (11S6169). Methods The DH population used in this study was developed by the following methods: we crossed normal corn (HF1) and normal corn (11S6169), which are parent lines of a normal corn cultivar, in 2014; and the next year, the F1hybrids were crossed with a tropicalized haploid inducer line (TAIL), which is homozygous for the dominant marker gene R1-nj (Nanda and Chase in Crop Sci 6:213–215, 1966), and we harvested seeds of the haploid lines. Results A total of 200 SSR markers were assigned to 10 linkage groups that spanned 1145.4 cM with an average genetic distance between markers of 5.7 cM. 68 SSR markers showed Mendelian segregation ratios in the DH population at a 5% significance threshold. A total of 15 quantitative trait loci (QTLs) for plant height (PH), ear height (EH), ear height ratio (ER), leaf length (LL), ear length (EL), set ear length (SEL), set ear ratio (SER), ear width (EW), 100 kernel weight (100 KW), and cob color (CC) were found in the 121 lines in the DH population. Conclusion The results of this study may help to improve the detection and characterization of agronomic traits and provide great opportunities for maize breeders and researchers using a DH population in maize breeding programs.

Phenotyping analysis of p53 knockout mice produced by gene editing and comparison with conventional p53 knockout mice Abstract Background Knockout (KO) mice developed by homologous recombination (HR) have become useful tools to elucidate gene function. However, HR has low KO efficiency and is time-consuming, labor-intensive, and expensive. 'Gene editing' has received much attention for efficient genetic manipulation. Objective As generation of KO mice is simplified, KO mice produced by HR can be feasibly reproduced using gene editing. However, phenotyping analysis and comparison between KO mice produced by these two techniques is necessary. Methods We generated p53 KO mice through gene editing and compared their phenotype with the already reported HR-mediated p53 KO mice. Results Tumors occurred in 36 (73%) of 49 homozygous KO mice and the mean age of occurrence was 23 weeks, with lymphoma (64%) and sarcoma (23%) being the most common. Tumors were also developed in 12 heterozygous mice and the mean age of occurrence was 40 weeks, with sarcoma (54%) and lymphoma (46%) in high proportion. Homozygotes had a mean life span of 157 ± 52 days and developmental abnormalities were found in females compared to in males (P < 0.05, P < 0.001). Conclusion We analyzed the basic phenotype of p53 KO mice and observed no significant difference from the conventional HR-mediated p53 KO mice.

Reversine induces caspase-dependent apoptosis of human osteosarcoma cells through extrinsic and intrinsic apoptotic signaling pathways Abstract Background The 2-(4-morpholinoanilino)-6-cyclohexylaminopurine (reversine) acts as a chemopreventive agent and induces apoptotic cell death in various cancer cells. However, the anticancer effects of reversine on osteosarcoma cells are not clearly established. Objective The purpose of this study was to investigate the effect of reversine on cell proliferation and induction of apoptosis in human osteosarcoma cells. Methods Cell viability assay, histological analysis, DAPI staining, caspase activation analysis, flow cytometric analysis and immunoblotting were carried out in MG-63 osteosarcoma cells. Results Reversine inhibited the growth of cells in a dose-dependent manner and induced nuclear condensation and fragmentation. Reversine-treated cells showed caspase-3/7 activation and increased apoptosis versus control cells. FasL, a death ligand associated with extrinsic apoptotic signaling pathways, was significantly up-regulated by reversine treatment. Moreover, the caspase-8, a part of the extrinsic apoptotic pathway, was activated by reversine treatments. Expressions of anti-apoptotic factors such as Bcl-2 and Bcl-xL, components of the mitochondria dependent intrinsic apoptosis pathway, significantly decreased following reversine treatment. The expressions of pro-apoptotic factors such as BAX, BAD and caspase-9 increased by reversine treatments. In addition, reversine activated caspase-3 and Poly (ADP-ribose) polymerase (PARP) to induce cell death. The Z-VAD-fmk significantly inhibited cell death through the suppression of caspase-3 expression in MG-63 cells treated with reversine. Conclusion These results suggest that the reversine may inhibit cell proliferation and induce apoptotic cell death in MG-63 osteosarcoma cells through both the mitochondria-mediated intrinsic pathway and the death receptor-mediated extrinsic pathway, and may have potential properties for the discovery of anti-cancer agents.

Transcriptome analysis for identifying possible causes of post-reproductive death of Sepia esculenta based on brain tissue Abstract Background The subpeduncle lobe/olfactory lobe–optic gland axis is called the endocrine regulation center of cephalopods. However, little is known about the mechanism of the subpeduncle lobe/olfactory lobe-optic gland axis regulate the sexual maturation and post-reproductive death of Sepia esculenta Hoyle. Objectives The primary objective of this study was to provide basic information for revealing the mechanism of the subpeduncle lobe/olfactory lobe–optic axis regulating the rapid post-reproductive death of S. esculenta. Methods In this paper, Illumina sequencing based transcriptome analysis was performed on the brain tissue of female S. esculenta in the three key developmental stages: growth stage (BG), spawning stage (BS), and post-reproductive death stage (BA). Results A total of 66.19 Gb Illumina sequencing data were obtained. A comparative analysis of the three stages showed 2609, 3333, and 170 differentially expressed genes (DEGs) in BG-vs-BA, BG-vs-BA, and BS-vs-BA, respectively. The Gene Ontology (GO) enrichment analysis of DEGs revealed that the regulation of cyclin-dependent protein serine/threonine kinase activity, oxidative phosphorylation, and respiratory chain were significantly enriched. The significant enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway identified pathways associated with the regulation of death, such as the mammalian target of rapamycin (mTOR) signaling pathway, AMPK signaling pathway, oxidative phosphorylation, and cell cycle. Conclusion The post-reproductive death of S. esculenta was found to be a complex energy steady-state regulation network system. The mTOR acted as an energy receptor and had a key role in regulating energy homeostasis.

Whole genome sequencing analysis of horse populations inhabiting the Korean Peninsula and Przewalski's horse Abstract Background The Jeju horse is an indigenous horse breed in Korea. However, there is a severe lack of genomic studies on Korean horse breeds. Objective The objective of this study was to report genomic characteristics of domestic horse populations that inhabit South Korea (Jeju, Jeju crossbred, and Thoroughbred) and a wild horse breed (Przewalski's horse). Results Using the equine reference genome assembly (EquCab 2.0), more than ~ 6.5 billion sequence reads were successfully mapped, which generated an average of 40.87-fold coverage throughout the genome. Using these data, we detected a total of 12.88 million SNPs, of which 73.7% were found to be novel. All the detected SNPs were deeply annotated to retrieve SNPs in gene regions using the RefSeq and Ensemble gene sets. Approximately 27% of the total SNPs were located within genes, whereas the remaining 73% were found in intergenic regions. Using 129,776 coding SNPs, we retrieved a total of 49,171 nonsynonymous SNPs in 12,351 genes. Furthermore, we identified a total of 10,770 deleterious nonsynonymous SNPs which are predicted to affect protein structure or function. Conclusion We showed numerous genomic variants from domestic and wild horse breeds. These results provide a valuable resource for further studies on functions of SNP-containing genes, and can aid in determining the molecular basis underlying variation in economically important traits of horses.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

PharmacoEconomics

Correction to: Venetoclax for Treating Chronic Lymphocytic Leukaemia: An Evidence Review Group Perspective of a NICE Single Technology Appraisal Open Access This article is distributed under the terms of the Creative Commons Attribution-Non Commercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Correction to: The Impact of Hospital Costing Methods on Cost-Effectiveness Analysis: A Case Study The Open Access license, which previously read.

Cost-Effectiveness Analysis in the Context of US Commercial Health Insurance

Pirfenidone for Treating Idiopathic Pulmonary Fibrosis: An Evidence Review Group Perspective of a NICE Single Technology Appraisal Abstract The National Institute for Health and Care Excellence (NICE) published guidance on the use of pirfenidone (Esbriet®, Roche) for the treatment of mild to moderate idiopathic pulmonary fibrosis (IPF) in 2013. NICE decided to review existing guidance following publication of an additional clinical trial, and invited the manufacturer of pirfenidone to submit evidence of its clinical and cost effectiveness for the treatment of mild to moderate IPF when compared with best supportive care (BSC) or nintedanib; nintedanib was a comparator only for moderate IPF. An independent Evidence Review Group (ERG) critiqued the company submission and this paper summarises their report and subsequent NICE guidance. The key clinical effectiveness evidence was based on three randomised controlled trials (RCTs) and an open-label extension study. Supportive data were provided from two additional RCTs conducted in Japan, while one additional open-label study was included for safety outcomes. Meta-analysis of the three key RCTs found pirfenidone to be effective at reducing disease progression compared with placebo, but statistically significant differences were not identified in all of the RCTs. A statistically significant reduction in all-cause mortality was only demonstrated when pooling data across studies. The treatment effects of pirfenidone and nintedanib were broadly similar, based on an indirect comparison using network meta-analysis, although they have slightly different adverse event profiles. There remains considerable uncertainty in the cost-effectiveness estimates for pirfenidone versus BSC, particularly due to uncertainty regarding the duration of treatment effect and the method used to implement the stopping rule within the economic model.

A Flexible Open-Source Decision Model for Value Assessment of Biologic Treatment for Rheumatoid Arthritis Abstract Objective The nature of model-based cost-effectiveness analysis can lead to disputes in the scientific community. We propose an iterative and collaborative approach to model development by presenting a flexible open-source simulation model for rheumatoid arthritis (RA), accessible to both technical and non-technical end-users. Methods The RA model is a discrete-time individual patient simulation with 6-month cycles. Model input parameters were estimated based on currently available evidence and treatment effects were obtained with Bayesian network meta-analysis techniques. The model contains 384 possible model structures informed by previously published models. The model consists of the following components: (i) modifiable R and C++ source code available in a GitHub repository; (ii) an R package to run the model for custom analyses; (iii) detailed model documentation; (iv) a web-based user interface for full control over the model without the need to be well-versed in the programming languages; and (v) a general audience web-application allowing those who are not experts in modeling or health economics to interact with the model and contribute to value assessment discussions. Results A primary function of the initial version of RA model is to help understand and quantify the impact of parameter uncertainty (with probabilistic sensitivity analysis), structural uncertainty (with multiple competing model structures), the decision framework (cost-effectiveness analysis or multi-criteria decision analysis), and perspective (healthcare or limited societal) on estimates of value. Conclusion In order for a decision model to remain relevant over time it needs to evolve along with its supporting body of clinical evidence and scientific insight. Multiple clinical and methodological experts can modify or contribute to the RA model at any time due to its open-source nature.

Fulvestrant for Untreated Hormone-Receptor Positive Locally Advanced or Metastatic Breast Cancer: An Evidence Review Group Perspective of a NICE Single Technology Appraisal Abstract Clinical and cost-effectiveness evidence on fulvestrant for untreated hormone-receptor positive locally advanced or metastatic breast cancer was submitted to the single technology appraisal process of the National Institute for Health and Care Excellence by the manufacturer of fulvestrant. The Southampton Health Technology Assessments Centre was commissioned by the National Institute for Health and Care Excellence as an independent Evidence Review Group to critique the company's submitted evidence. Fulvestrant was compared directly with anastrozole in two randomised controlled trials and was compared indirectly by means of a network meta-analysis with anastrozole, letrozole and tamoxifen. This article summarises the Evidence Review Group's review of the company's submission and summarises the guidance the National Institute for Health and Care Excellence Appraisal Committee issued in January 2018. The Evidence Review Group had several concerns, the most important of which related to the degree to which fulvestrant might confer a benefit in overall survival. This was because mature data were not available from the key phase III trial FALCON. The economic model was sensitive to changes in overall survival and the Evidence Review Group considered the incremental cost-effectiveness ratio was uncertain and likely to increase once mature results from FALCON become available. The National Institute for Health and Care Excellence Appraisal Committee concluded that fulvestrant could not be recommended for treating locally advanced or metastatic estrogen-receptor-positive breast cancer in postmenopausal women who have not received previous endocrine therapy.

Estimation of Cost for Endoscopic Screening for Esophageal Cancer in a High-Risk Population in Rural China: Results from a Population-Level Randomized Controlled Trial Abstract Background and Objective Population-level endoscopic screening for esophageal cancer has been conducted in China for years. In this study, we aim to provide an updated and precise cost estimation for esophageal cancer screening based on a randomized controlled trial in a high-risk area in China. Methods We estimated the cost of esophageal cancer screening with chromoendoscopy using a micro-costing approach based on primary data of the ESECC (Endoscopic Screening for Esophageal Cancer in China) randomized controlled trial (NCT01688908) from a health sector perspective. Unit costs and quantities of resources were collected to obtain annual screening costs. The screening project was then theoretically expanded to a 10-year period to explore long-term trends of costs. Costs were adjusted to US dollars for the year 2018. Results In the ESECC trial, screening cost per endoscopy with a valid pathologic diagnosis was $196, accounting for 3.82% of the gross domestic product per capita in Hua County, and the costs for detecting one esophageal cancer and one early-stage esophageal cancer were $26,347 and $37,687, respectively. In conventional screening in which protocol-driven costs were excluded, costs as above were $134, $18,074, and $25,853. The cost for detecting one gastric cardia cancer or stomach cancer was nine times higher than detecting one esophageal cancer owing to low prevalences of the two cancers. In a simulated 10-year screening project, annual cost decreased notably over time. Conclusions Despite the relatively low absolute cost, population-level endoscopic screening will still be a heavy burden on local government considering the socioeconomic conditions. Long-lasting programs would be less costly and population-level screening would make little sense in non-high-risk regions.

Cost Effectiveness of Nusinersen in the Treatment of Patients with Infantile-Onset and Later-Onset Spinal Muscular Atrophy in Sweden Abstract Background Spinal muscular atrophy is a rare neuromuscular disorder with a spectrum of severity related to age at onset and the number of SMN2 gene copies. Infantile-onset (≤ 6 months of age) is the most severe spinal muscular atrophy and is the leading monogenetic cause of infant mortality; patients with later-onset (> 6 months of age) spinal muscular atrophy can survive into adulthood. Nusinersen is a new treatment for spinal muscular atrophy. Objective The objective of this study was to evaluate the cost effectiveness of nusinersen for the treatment of patients with infantile-onset spinal muscular atrophy and later-onset spinal muscular atrophy in Sweden. Methods One Markov cohort health-state transition model was developed for each population. The infantile-onset and later-onset models were based on the efficacy results from the ENDEAR phase III trial and the CHERISH phase III trial, respectively. The cost effectiveness of nusinersen in both models was compared with standard of care in Sweden. Results For a time horizon of 40 years in the infantile-onset model and 80 years in the later-onset model, treatment with nusinersen resulted in 3.86 and 9.54 patient incremental quality-adjusted life-years and 0.02 and 2.39 caregiver incremental quality-adjusted life-years and an incremental cost of 21.9 and 38.0 million SEK (Swedish krona), respectively. These results translated into incremental cost-effectiveness ratios (including caregiver quality-adjusted life-years) of 5.64 million SEK (€551,300) and 3.19 million SEK (€311,800) per quality-adjusted life-year gained in the infantile-onset model and later-onset model, respectively. Conclusions Treatment with nusinersen resulted in overall survival and quality-adjusted life-year benefits but with incremental costs above 21 million SEK (€2 million) [mainly associated with maintenance treatment with nusinersen over a patient's lifespan]. Nusinersen was not cost effective when using a willingness-to-pay threshold of 2 million SEK (€195,600), which has been considered in a recent discussion by the Dental and Pharmaceutical Benefits Agency as a reasonable threshold for rare disease. Nonetheless, nusinersen gained reimbursement in Sweden in 2017 for paediatric patients (below 18 years old) with spinal muscular atrophy type I–IIIa.

Cost Effectiveness of Sodium-Glucose Cotransporter-2 (SGLT2) Inhibitors, Glucagon-Like Peptide-1 (GLP-1) Receptor Agonists, and Dipeptidyl Peptidase-4 (DPP-4) Inhibitors: A Systematic Review Abstract Objective This study aimed to systematically review cost-effectiveness studies of newer antidiabetic medications. Methods The PubMed/MEDLINE, EMBASE, CINAHL Plus, Cochrane Library–NHS Economic Evaluation Database (Wiley), Cochrane Library–Health Technology Assessment Database (Wiley), Cochrane Library–Database of Abstracts of Reviews of Effects (Wiley), and the Cost-Effectiveness Analysis Registry databases (from 1 January 2000 to 1 June 2018) were searched. The search strategies included the Medical Subject Heading (MeSH) term 'economics', and the MeSH entry terms 'cost', 'cost effectiveness', 'value', and 'cost utility', as well as all names for GLP-1 receptor agonists, DPP-4 inhibitors, and SGLT2 inhibitors. Inclusion criteria included (1) cost-effectiveness studies of the newer antidiabetic medications, including sodium-glucose cotransporter-2 (SGLT2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, and dipeptidyl peptidase-4 (DPP-4) inhibitors; and (2) full-text publications in English. Two reviewers independently screened the titles, abstracts, and full-text articles to select studies for data extraction. Discrepancies were resolved by discussion and consensus. The quality of reporting cost-effectiveness analyses was assessed using the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) guideline. Results Among 85 studies selected, 82 clearly stated the types of diabetes model used (e.g. CORE model), and 70 studied used validated diabetes models. Seventy-four (87%) studies were funded by pharmaceutical companies, and 72 (85%) studies were conducted from a payer's perspective. Seventy-six (89%) studies presented were of good quality (20–24 CHEERS items), and nine were of moderate quality (14–19 items). Thirty studies compared newer antidiabetic medications with insulin, 3 studies compared newer antidiabetic medications with thiazolidinediones (TZDs), 15 studies compared newer antidiabetic medications with sulfonylureas, 40 studies compared new antidiabetic medications with alternative newer antidiabetic medication, and 9 studies compared other antidiabetic agents that were not included above. Newer antidiabetic medications were reported to be cost-effective in 26 of 30 (87%) studies compared with insulin, and 13 of 15 (87%) studies compared with sulfonylureas. Conclusions Most economic evaluations of antidiabetic medications have good reporting quality and use validated diabetes models. The newer antidiabetic medications in most of the reviewed studies were found to be cost effective, compared with insulin, TZDs, and sulfonylureas.

Combining the Power of Artificial Intelligence with the Richness of Healthcare Claims Data: Opportunities and Challenges Abstract Combinations of healthcare claims data with additional datasets provide large and rich sources of information. The dimensionality and complexity of these combined datasets can be challenging to handle with standard statistical analyses. However, recent developments in artificial intelligence (AI) have led to algorithms and systems that are able to learn and extract complex patterns from such data. AI has already been applied successfully to such combined datasets, with applications such as improving the insurance claim processing pipeline and reducing estimation biases in retrospective studies. Nevertheless, there is still the potential to do much more. The identification of complex patterns within high dimensional datasets may find new predictors for early onset of diseases or lead to a more proactive offering of personalized preventive services. While there are potential risks and challenges associated with the use of AI, these are not insurmountable. As with the introduction of any innovation, it will be necessary to be thoughtful and responsible as we increasingly apply AI methods in healthcare.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Plastic Surgery

Correction to: Optimization of the dermal wrap in inferior pedicle reduction mammoplasty: An Egyptian experience The authors wish to clarify that Dina M. Badwai refers to Dina M. Badawi. The authors apologise for this error.

Management of abnormal deep inferior epigastric vessels in microsurgical breast reconstruction: a report of 3 complex cases Abstract The deep inferior epigastric artery perforator flap is the gold standard in microsurgical breast reconstruction. It is now increasingly offered in patients with previous abdominal scars and in such cases, computed tomography angiography (CTA) is routinely implemented. However, previous injury of the deep inferior epigastric vessels may be difficult to detect with CTA. We report three cases of successful bilateral breast reconstruction, in which a unilateral DIEA was found to be abnormal. In the first two cases, the unilateral DIE vessels were found to be damaged and the flaps salvaged based on the deep superior epigastric artery. In the third patient, previous DIEA injury resulted in vessel recanalization, which was only evident following initial anastomotic failure. In this case, the damaged DIEA segment was subsequently removed and the flap successfully revascularized. All three patients had an uneventful post-operative recovery and all flaps survived with no complications. Based on our experience, we believe that awareness of the potential pitfalls and careful planning for lifeboats are essential to avoid flap loss in such circumstances. Level of Evidence: Level V, therapeutic study.

Surgical fire: principles, risk factors, and prevention

Trigger finger in a hereditary multiple exostoses disease: A unique case report Abstract Trigger finger is one of the most common upper extremity problems seen by hand surgeons. Lesions occupying space in the tendon bed can prevent tendon gliding. We describe a unique case of trigger finger in a patient known for a hereditary multiple exostoses disease where an exostosis in the tendon bed constricted the tendon sheath, leading to triggering and locking. Open surgical treatment was performed by removing of the exostosis which relieved the problem. Level of Evidence: Level V, therapeutic study.

Forehead biconvexity enhancement with fat grafting Abstract Background Frontal biconvexity is a key criterion for an attractive forehead. Fat injection as an effective, safe and reliable method for soft tissue augmentation could be used to enhance forehead contour. We report our experience with combined platelet-rich plasma (PRP) and fat grafting to create or restore frontal biconvexity and to reduce wrinkles. Methods Fifty-seven females and 4 males (mean age, 40.2 years) underwent the combined PRP and fat injection. Fat mixed with PRP on the ratio of 5:1 was injected in small aliquots in the space between the dermis and underlying frontalis muscle. Patient satisfaction with the cosmetic result was evaluated by a questionnaire using pre- and postoperative photos and a four-point grading scale. Results All patients who underwent three injection sessions (n = 5) evaluated the esthetic result as excellent. In patients who had two injection sessions (n = 36), 36% evaluated the result as excellent, 50% as good, and the remaining 5 as moderate. In patients who had a single injection session (n = 15), 27% evaluated the result as excellent, 40% as good, and the remaining 5 as moderate. Concerning frontal wrinkles, 25 of 34 (73.5%) patients reported good improvement and the remaining 9 (26.5%) reported moderate improvement. Conclusion Fat injection combined with PRP should be considered as an effective tool to improve forehead biconvexity. Level of evidence: Level IV, therapeutic study.

Pretarsal roll augmentation with dermal hyaluronic acid filler injection Abstract Pretarsal roll augmentation with dermal hyaluronic acid filler injection focuses on restoring pretarsal fullness. This study aimed to introduce a method of pretarsal roll augmentation with dermal hyaluronic acid filler injection and establish the level of difficulty, safety, and effectiveness of this method. Eighty female patients were enrolled in this study. Hyaluronic acid filler was used to perform pretarsal roll augmentation. Physician and patient satisfaction at 1 month and 4 months after surgery was investigated. The level of satisfaction was graded from points 1 to 5. The patient satisfaction and physician scores were 4.7 ± 1.1 (mean ± standard deviation) points at 1 month and 4.8 ± 0.9 points at 4 months and 4.6 ± 0.9 points at 1 month and 4.8 ± 1.0 points at 4 months, respectively. No major complications were observed. Our technique provided a natural and younger appearance with pretarsal fullness. This technique was easy to perform for the restoration of pretarsal fullness, and it improved periorbital contouring, rejuvenated the pretarsal roll, and provided excellent esthetic results. Level of Evidence: Level V, therapeutic study.

An anatomical study of vascular communications between anterior tibial and peroneal osseosomes and its clinical application in proximal hemiarthroplasty of radiocarpal joint following tumor excision Abstract Background There are speculations that the proximal third fibular osseosomes, supplied by the anterior tibial vessels cannot be reliably recruited on the peroneal vessels supplying fibular diaphyseal osseosomes. Anatomical study by the authors eschewed this speculation and paved way for an evidence-based surgery. Cadaveric dissection, dye injection, and radiological studies revealed existence of a reliable anastomotic musculoperiosteal vessel between these two osseosomes that could effectively recruit the proximal fibular osseosomes on the peroneal vessels. Also, operative tips and tricks for safeguarding these anastomotic vessels were defined by the cadaveric study. Aesthetic and functional outcomes were assessed in five clinical cases, where the fibular heads were harvested reliably on the peroneal pedicle and stable wrist joints were reconstructed following oncological excisions. Methods Based on the evidence from the anatomical study, five clinical cases (three males and two females) affected by stage III GCT (giant cell tumor) of distal radius underwent microvascular fibular head hemiarthroplasty following tumor excision. Average follow-up of the patients was 18 months. Results At the end of follow-up period, all patients had an average MSTS (Musculoskeletal Tumor Society) score of 23. Scores ranged from 21 to 25. Patients had good functional outcomes with 69% average mobility in the reconstructed wrist when compared with the opposite normal wrist. Aesthesis of the reconstructed wrists were acceptable. Conclusion The cadaveric study demonstrated concrete anatomical evidence of existence of an anastomotic musculoperiosteal vessel between anterior tibial and peroneal fibular osseosomes in all the specimens. Clinical study validated the safety of the vascularized proximal fibular hemiarthroplasty on peroneal pedicle. It may be a reliable method for reconstruction of the radiocarpal joint in the post-GCT distal radius excision with good aesthetic and functional outcomes. Level of Evidence: Level V, therapeutic study

Morbidity and mortality in patients undergoing fecal diversion as an adjunct to wound healing: a NSQIP comparison study Abstract Background Fecal diversion for chronic, non-healing wounds improves quality of life, assists in wound healing, and helps to prepare for reconstructive surgery. While commonplace, little has been published regarding the safety of diversion in this patient subgroup. The purpose of this study is to elucidate the morbidity and mortality of fecal diversion for chronic wounds and to identify those patients with disproportionately high perioperative risk. Methods Retrospective analyses were performed using the American College of Surgeons National Surgical Quality Improvement Project (NSQIP) database and an institutional database. The primary outcome analyzed was 30-day mortality and secondary outcomes included 30-day morbidity and readmission rate. Results Eight hundred fifty-nine patients were identified in the NSQIP database who underwent diversion compared to 3990 who did not. In unmatched data, there were no significant differences in substantial 30-day morbidities. In matched data, diverted patients had a significantly lower perioperative mortality. Fifty-six patients were identified in the institutional review who were diverted for non-healing wounds. Fifty percent of patients with a preoperative ejection fraction of less than 30% died within 30 days of surgery (LR 6.58, p = 0.045). Conclusions The NSQIP review indicates that fecal diversion does not inherently increase 30-day perioperative morbidity or mortality. While 30-day morbidity remains high, the institutional review suggests that patients with cardiac dysfunction contribute to the majority of complications. As such, an ejection fraction less than 30% may be a relative contraindication to immediate diversion. Medical optimization and elective diversion should be considered whenever feasible. Level of Evidence: Level III, risk / prognostic study.

Optimization of the dermal wrap in inferior pedicle reduction mammoplasty: An Egyptian experience Abstract Background Reduction mammoplasty using the inferior pedicle technique has been used for more than five decades. Without doubt, it has many advantages, especially in patients with moderate to severe macromastia. Although the esthetic outcome of reduction mammoplasty in the early postoperative period is usually satisfactory, typical esthetic drawbacks appear during the first year. These drawbacks include bottoming out, loss of superior fullness, box-shaped breasts, poor definition of the infra-mammary fold (IMF), elongation of the vertical scar, star gazing of the nipple-areola complex (NAC) and, of course, the scar burden. Parenchymal reshaping and suspension have been introduced as an additional step to eliminate the drawbacks of the inferior pedicle procedure. Methods This is a prospective study done in a period from March 2016 to March 2018 in Kasr Al-Ainy Hospital (Cairo University hospitals) by the senior author and his team of co-authors. Reduction mammoplasty (Wise pattern) with the inferior pedicle and dermal wrap technique (using two dermal wings) was performed in 20 patients. The distance from the suprasternal notch (SSN) to the NAC was > 30 cm. Results The patients' ages ranged from 28 to 55 years (average age, 39 years). Body mass index (BMI) ranged from 27 to 35 kg/m2 (average BMI, 33.3 kg/m2). The distance between the SSN and NAC ranged from 33 to 45 cm (average distance, 40 cm). The nipples were lifted by 11 to 19.5 cm. The distance from the nipple to the IMF ranged from 18 to 24 cm. This distance was reduced postoperatively to between 9 and 11.5 cm (p value < 0.001). Conclusions Follow-up of the patients revealed that suspension of the breast parenchyma and plication of the dermal flap improved the esthetic results following inferior pedicle breast reduction. The addition of volumetric and geometric adjustment "preoperatively" may significantly improve the results and minimize the operative time of this relatively lengthy procedure (4–6 h for both breasts). Level of Evidence: Level IV, terapeutic study.

Therapeutic breast reduction—are doctors and patients satisfied ? Abstract Background Therapeutic breast reduction (TBR) is an oncoplastic technique that applies breast reduction principles for oncologic purposes. Given that TBR indications have expanded, the purpose of this study is to ascertain the aesthetic outcome of this procedure, and determine how it may be influenced by different surgical techniques and postoperative radiotherapy. Methods A non-randomized cohort study was performed, including breast cancer female patients who underwent breast conserving surgery with TBR. The primary outcome was the esthetic result of the reconstruction, evaluated by both plastic surgeons and patients, at least 12 months after surgery. Results The aesthetic assessment was made in 42 patients. Overall, the clear majority of patients classified the outcome as good or perfect (95.2%), with less than 5% considering the outcome as mediocre. As for plastic surgeons, 83.3% were considered perfect/good outcomes, with 16.7% mediocre results. There were no poor results, neither for the patient nor the surgeon. There were no statistically significant associations between the esthetic result and tumor location nor its relation to the skin-resection pattern. Conclusion Aesthetic outcomes with this technique are promising, even when there is the need for technical modifications and despite the need for adjuvant radiotherapy, making it valuable for tumors in all locations. Level of Evidence: Level III, risk / prognostic study.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Environmental Science and Pollution Research

Estimation of methane and nitrous oxide emission from wetland rice paddies with reference to global warming potential Abstract Methane (CH4) and nitrous oxide (N2O) are two important greenhouse gases (GHG) and contribute largely to global warming and climate change. The impact of physiological characteristics of rice genotypes on global warming potential (GWP) and greenhouse gas intensity (GHGI) is not well documented. A 2-year field experiment was conducted with eight summer rice varieties: Dinanath, Joymoti, Kanaklata, Swarnabh, IR 64, Tapaswami (modern varieties), Number 9, and Jagilee Boro (indigenous varieties) for two successive seasons (December–June, 2015–2016 and December–June, 2016–2017) to estimate their GWP and GHGI. The GWP of the rice varieties ranged from 841.52 to 1288.67 kg CO2-equiv. ha−1 and GHGI from 0.184 to 0.854 kg CO2-equiv. kg−1 grain yield. Significant differences (p < 0.05) in seasonal GHG emission, GWP, GHGI, CEE (carbon equivalent emission), photosynthetic efficiency, stomatal conductance, transpiration rate, and grain productivity among the rice varieties were observed during the investigation. A good correlation of GWP (p < 0.01) was recorded with rate of stomatal conductance and transpiration rate of the varieties. The present study reveals a strong relationship between plant biomass (p < 0.01) with GWP and CEE of the rice varieties. The variety IR 64 and Number 9 are identified as the most suitable variety with lowest GWP (909.85 and 876.68 kg CO2-equiv. ha−1 respectively) and GHGI (0.192 and 0.227 kg CO2-equiv. kg−1 grain yield respectively) accompanied by higher grain productivity (4839 and 3867 kg ha−1 respectively). Observations from the study suggest that agricultural productivity and GHG mitigation can be simultaneously achieved by proper selection of rice genotypes.

Optimisation of bioscrubber systems to simultaneously remove methane and purify wastewater from intensive pig farms Abstract The use of bioscrubber is attracting increasing attention for exhaust gas treatment in intensive pig farming. However, the challenge is to improve the methane (CH4) removal efficiency as well as the possibility of pig house wastewater treatment. Three laboratory-scale bioscrubbers, each equipped with different recirculation water types, livestock wastewater (10-times-diluted pig house wastewater supernatant), a methanotroph growth medium (10-times-diluted), and tap water, were established to evaluate the performance of CH4 removal and wastewater treatment. The results showed that enhanced CH4 removal efficiency (25%) can be rapidly achieved with improved methanotrophic activity due to extra nutrient support from the wastewater. The majority of the CH4 was removed in the middle to end part of the bioscrubbers, which indicated that CH4 removal could be potentially optimised by extending the length of the reactor. Moreover, 52–86% of the ammonium (NH4+-N), total organic carbon (TOC), and phosphate (PO43−-P) removal were simultaneously achieved with CH4 removal in the present study. Based on these results, this study introduces a low-cost and simple-to-operate method to improve CH4 removal and simultaneously treat pig farm wastewater in bioscrubbers.

Ambient air pollution and risk of type 2 diabetes in the Chinese Abstract We performed a time series analysis to investigate the potential association between exposure to ambient air pollution and type 2 diabetes (T2D) incidence in the Chinese population. Monthly time series data between 2008 and 2015 on ambient air pollutants and incident T2D (N = 25,130) were obtained from the Environment Monitoring Center of Ningbo and the Chronic Disease Surveillance System of Ningbo. Relative risks (RRs) and 95% confidence intervals (95% CIs) of incident T2D per 10 μg/m3 increases in ambient air pollutants were estimated from Poisson generalized additive models. Exposure to particulate matter < 10 μm (PM10) and sulfur dioxide (SO2) was associated with increased T2D incidence. The relative risks (RRs) of each increment in 10 μg/m3 of PM10 and SO2 were 1.62 (95% CI, 1.16–2.28) and 1.63 (95% CI, 1.12–2.38) for overall participants, whereas for ozone (O3) exposure, the RRs were 0.78 (95% CI, 0.68–0.90) for overall participants, 0.78 (95% CI, 0.69–0.90) for males, and 0.78 (95% CI, 0.67–0.91) for females, respectively. Exposure to PM10 and SO2 is positively associated with T2D incidence, whereas O3 is negatively associated with T2D incidence.

The relief effects of organic acids on Scirpus triqueter L. under pyrene–lead stress Abstract During phytoremediation of polycyclic aromatic hydrocarbons (PAHs) and heavy metals, the phytoremediation plants are often stressed by pollutants, which would reduce the efficiency of phytoremediation. The addition of organic acids from root exudates could alleviate the stress. In this study, three organic acids (citric acid, succinic acid, glutaric acid) were added to investigate the effects of organic acids on the stress response of Scirpus triqueter L. at two pyrene–lead concentrations. The activities of reactive oxygen species, malondialdehyde, plasma membrane H+-ATPase, and vacuolar H+-ATPase and PPase activity, as well as the activities of antioxidant enzymes (SOD, POD, and CAT) in Scirpus triqueter L. were determined. The addition of organic acids could effectively reduce the activities of reactive oxygen species, malondialdehyde, plasma membrane H+-ATPase, and vacuolar H+-ATPase and PPase activities. Under higher pollution, the damage of plant plasma membrane is more serious, but the addition of citric acid can alleviate this situation and even more effective than the relief under low pollution. The effect of citric acid was more significant than that of succinic acid and glutaric acid. These results demonstrated that organic acids could attenuate the stress of pyrene and lead to Scirpus triqueter L.

The release mechanism of heavy metals from lab-scale vertical flow constructed wetlands treating road runoff Abstract Constructed wetlands (CWs) have been applied to remediate heavy metal pollution effectively in practice. However, the heavy metal release from CWs has not been paid enough attention. In this study, a 5-month experiment was carried out with three parallel lab-scale vertical flow constructed wetlands (VFCWs) with zeolites as fillers. The artificial rainwater was pumped into VFCWs to study the release characteristic and mechanisms of heavy metals (Cu, Zn, Cr, and Pb). The results showed that significant amounts of Zn and Cu were released from the VFCWs at the end of the experiment while Pb and Cr rarely escaped. The upper layer (0–30 cm) of the VFCWs was the most effective area for heavy metal removal due to the presence of sediments, but it was also the most active area for heavy metal release. To explain this result, the sediments were analyzed before and after being leached by the tap water. The results indicated that Zn and Cu existed mainly in the exchangeable state, and they had strong leachability and bioavailability, causing its releases. Also, competitive adsorption of different metals meant that the metal ions with strong adsorption to zeolite caused the metal ions with weak adsorption to be desorbed from zeolites, and thus, a large amount of Zn escaped from VFCWs. The escape of heavy metals from CWs illustrated that it should be paid more attention in the management.

Concentration, spatiotemporal distribution, and sources of mercury in Mt. Yulong, a remote site in southeastern Tibetan Plateau Abstract The unique geographic location of Mt. Yulong in the Tibetan Plateau (TP) makes it a favorable site for mercury (Hg) study. Various snow samples, such as surface snow, snow pit, and snowmelt water were collected from Mt. Yulong in the southeastern TP. The average concentration of Hg was found to be 37 ± 26 ng L−1 (mean ± SD), comparable to Hg concentration from other parts of TP in the same year, though it was comparatively higher than those from previous years, suggesting a possible increase of Hg concentration over the TP. The concentration of Hg was higher in the lower elevation of the glaciers possibly due to the surface melting concentration of particulates. Higher concentration of Hg was observed in the fresh snow, suggesting the possibility of long-range transportation. The average concentration of Hg from the snow pit was 1.49 ± 0.78 ng L−1, and the concentration of Hg in the vertical profile of the snow pit co-varied with calcium ion (Ca2+) supporting the fact that the portion of Hg is from the crustal origin. In addition, the principal component analysis (PCA) confirmed that the source of Hg is from the crustal origin; however, the presence of anthropogenic source in the Mt. Yulong was also observed. In surface water around Mt. Yulong, the concentration of HgT was found in the order of Lashihai Lake > Reservoirs > Rivers > Swamps > Luguhu Lake. In lake water, the concentration of HgT showed an increasing trend with depth. Overall, the increased concentration of Hg in recent years from the TP can be of concern and may have an adverse impact on the downstream ecosystem, wildlife, and human health.

Evaluation of management procedures for a length-frequency data-limited fishery Abstract Management procedures (MPs) based on data-limited methods (DLMs) recently developed to give management advices for data-limited stocks worldwide are scarce or yet to be implemented on freshwater species. In this study, case studies (CSs) were developed using length-frequency data (LFD) of common carp species harvested from Dianshan Lake to estimate life-history parameters from existing methods. These CSs were later used to examine their influences when tested with various MPs under scenarios when operating models (OMs) were subjected to observation and estimation uncertainties. The results after management strategy evaluation (MSE) was run for various defined OMs showed that three MPs emerged best for providing managing advice. For high yield to be maintained during short-term periods, MinlenLopt1 suggested the smallest length at full retention (sLFR) to be 42.11 cm; while Slotlim and matlenlim2 suggested that to maintain biomass and stable spawning biomass (SBMSY) and also avoid overfishing from occurring in this fishery, sLFR should be 56.1 cm. Values given by these MPs allowed the removal of species that spawned at least once. Also, life-history parameters derived from CS4 presented the best results, being more reliable in presenting better inputs for effective management of the said fishery.

Remediation of cobalt-polluted soil after application of selected substances and using oat ( Avena sativa L.) Abstract The aim of the study was to determine the effectiveness of soil application of manure, clay, charcoal, zeolite, and calcium oxide in remediation of soil polluted with cobalt (0, 20, 40, 80, 160, 320 mg Co kg−1 of soil). The following were determined: weight of harvested plants as well as the content of cobalt in grain, straw, and roots of oat. In addition, tolerance index (Ti), cobalt bioconcentration (BCF), translocation (TF), and transfer (TFr) coefficients were derived. In the series without amendments, the increasing doses of cobalt had a significant effect by decreasing the yields of oat grain and straw and the mass of its roots. Also, lower tolerance index values were noted in the objects polluted with cobalt, especially with its highest dose. The application of manure had the strongest effect on increasing the mass of particular organs of the test plant, while the application of charcoal led to a significant decrease in this respect. The application of all substances to the soil, and especially manure and calcium oxide, resulted in higher tolerance index Ti values. The growing contamination of soil with cobalt caused a significant increase in the content of this element in oat and in the values of the translocation coefficient, in contrast to the effects noted with respect to the bioconcentration and transfer coefficients. All the substances applied to soil reduced the content of cobalt and its bioconcentration in oat straw, in opposition to grain and roots, limited its translocation, but elevated the transfer of this element from soil to plants. Soil contamination with cobalt promoted the accumulation of lead and copper in grain, cadmium, lead, nickel, zinc, manganese, and iron in straw, as well as cadmium, nickel, zinc, and manganese in oat roots. As the cobalt dose increased, the content of other trace elements in oat organs either decreased or did not show any unambiguous changes. Of all the tested substances, the strongest influence on the content of trace elements was produced by calcium oxide in straw and roots and by zeolite in roots, whereas the weakest effect was generated by manure in oat grain. Oat is not the best plant for phytoremediation of soils contaminated with cobalt.

Target and non-target botanical pesticides effect of Trichodesma indicum (Linn) R. Br. and their chemical derivatives against the dengue vector, Aedes aegypti L. Abstract The effects of crude ethanol derived leaf extract Trichodesma indicum (Linn) (Ex-Ti) and their chief derivatives were accessed on the survival and development of the dengue mosquito Ae. aegypti also their non-toxic activity against mosquito predator. T. indicum is recognized to be the vital weed plant and a promising herb in the traditional ayurvedic medicine. In this study, the GC-MS chromatogram of Ex-Ti showed higher peak area percentage for cis-10-Heptadecenoic acid (21.83%) followed by cycloheptadecanone (14.32%). The Ex-Ti displayed predominant mortality in larvae with 96.45 and 93.31% at the prominent dosage (200 ppm) against III and IV instar. Correspondingly, sub-lethal dosage against the enzymatic profile of III and IV instar showed downregulation of α,β-carboxylesterase and SOD protein profiles at the maximum concentration of 100 ppm. However, enzyme level of GST as well as CYP450 increased significantly dependent on sub-lethal concentration. Likewise, fecundity and hatchability of egg rate of dengue mosquito decreased to the sub-lethal concentration of Ex-Ti. Repellent assay illustrates that Ex-Ti concentration had greater protection time up to 210 min at 100 ppm. Also, activity of Ex-Ti on adult mosquito displayed 100% mortality at the maximum dosage of 600, 500 and 400 ppm within the period of 50, 60 and 70 min, respectively. Photomicrography screening showed that lethal dosage of Ex-Ti (100 ppm) produced severe morphological changes with dysregulation in their body parts as matched to the control. Effects of Ex-Ti on the Toxorhynchites splendens IV instar larvae showed less mortality (43.47%) even at the maximum dosage of 1500 ppm as matched to the chemical pesticide Temephos. Overall, the present research adds a toxicological valuation on the Ex-Ti and their active constituents as a larvicidal, repellent and adulticidal agents against the global burdening dengue mosquito.

Integrated lipid production, CO 2 fixation, and removal of SO 2 and NO from simulated flue gas by oleaginous Chlorella pyrenoidosa Abstract CO2, SO2, and NO are the main components of flue gas and can cause serious environmental issues. Utilization of these compounds in oleaginous microalgae cultivation not only could reduce air pollution but could also produce feedstock for biodiesel production. However, the continuous input of SO2 and NO inhibits microalgal growth. In this study, the toxicity of simulated flue gas (15% CO2, 0.03% SO2, and 0.03% NO, balanced with N2) was reduced through automatic pH feedback control. Integrated lipid production and CO2 fixation with the removal of SO2 and NO was achieved. Using this technique, a lipid content of 38.0% DW was achieved in Chlorella pyrenoidosa XQ-20044. The lipid composition and fatty acid profile indicated that lipid production by C. pyrenoidosa XQ-20044 cultured with flue gas is suitable as a biodiesel feedstock; 81.2% of the total lipids were neutral lipids and 99.5% of the total fatty acids were C16 and C18. The ratio of saturated fatty acids to monounsaturated fatty acids in the microalgal lipid content was 74.5%. In addition, CO2, SO2, and NO from the simulated flue gas were fixed and converted to biomass and lipids with a removal efficiency of 95.9%, 100%, and 84.2%, respectively. Furthermore, the utilization efficiencies of CO2, SO2, and NO were equal to or very close to their removal efficiencies. These results provide a novel strategy for combining biodiesel production with biofixation of flue gas.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Phonetics

Statistical distributions of consonant variants in infant-directed speech: Evidence that /t/ may be exceptional Publication date: July 2019 Source: Journal of Phonetics, Volume 75 Author(s): Laura Dilley, Jessica Gamache, Yuanyuan Wang, Derek M. Houston, Tonya R. Bergeson Abstract Statistical distributions of phonetic variants in spoken language influence speech perception for both language learners and mature users. We theorized that patterns of phonetic variant processing of consonants demonstrated by adults might stem in part from patterns of early exposure to statistics of phonetic variants in infant-directed (ID) speech. In particular, we hypothesized that ID speech might involve greater proportions of canonical /t/ pronunciations compared to adult-directed (AD) speech in at least some phonological contexts. This possibility was tested using a corpus of spontaneous speech of mothers speaking to other adults, or to their typically-developing infant. Tokens of word-final alveolar stops – including /t/, /d/, and the nasal stop /n/ – were examined in assimilable contexts (i.e., those followed by a word-initial labial and/or velar); these were classified as canonical, assimilated, deleted, or glottalized. Results confirmed that there were significantly more canonical pronunciations in assimilable contexts in ID compared with AD speech, an effect which was driven by the phoneme /t/. These findings suggest that at least in phonological contexts involving possible assimilation, children are exposed to more canonical /t/ variant pronunciations than adults are. This raises the possibility that perceptual processing of canonical /t/ may be partly attributable to exposure to canonical /t/ variants in ID speech. Results support the need for further research into how statistics of variant pronunciations in early language input may shape speech processing across the lifespan.

Spontaneous nasalization after glottal consonants in Thai Publication date: July 2019 Source: Journal of Phonetics, Volume 75 Author(s): Sarah E. Johnson, Marissa Barlaz, Ryan K. Shosted, Brad P. Sutton Abstract Spontaneous nasalization is the emergence of distinctive nasalization in contexts lacking an historical etymological nasal. In Thai, low and mid-low vowels nasalize after /h/ and to a lesser degree after /ʔ/. It has been reasoned that nasalization after /h/ may occur because breathiness and nasalization are acoustically similar; both introduce higher energy at low frequencies and increase spectral tilt. Glottal consonants may generally facilitate nasalization because aerodynamically they do not require velopharyngeal closure. We investigated velopharyngeal opening (VPO) during vowels after /h/ and/ ʔ/ and measured spectral tilt (H1–H2). We measured VPO by processing oblique ultra-fast magnetic resonance images of the velopharyngeal port. Four Thai speakers exhibited a complex system of VPO that varied based on vowel height and preceding consonant. Low vowels after /h/ manifested more physiological nasalization than low vowels after /ʔ/, while the former were often produced with higher spectral tilt, which may be indicative of either increased breathiness or nasalization. While VPO is likely responsible for impressions of greater nasalization after /h/, our findings suggest that breathiness and VPO may interact in the spontaneously nasalized vowels of Thai.

Influence of coda stop features on perceived vowel duration Publication date: July 2019 Source: Journal of Phonetics, Volume 75 Author(s): Chelsea Sanker Abstract Four experiments tested what cues contribute to English speakers' perception of vowel duration. Listeners categorized the duration of vowels as 'long' or 'short' for stimuli produced with voiced, voiceless, breathy voiced, or voiceless aspirated stop codas. Listeners demonstrated a strong ability to perceive vowel duration, though perception was continuous rather than categorical. There were several interacting factors influencing perceived vowel duration, based on expectations set by the presence of particular codas and also acoustic effects of the coda on the vowel. When the coda was removed, vowels that had been produced before voiced codas were perceived as longer than vowels produced before voiceless codas, though they exhibited the opposite effect when codas were present. Vowels were also perceived as longer when produced before breathy voiced stops, regardless of whether or not the stop was present. The steeper f0 falls associated with voiced codas within these stimuli likely contributed to the longer perceived duration of vowels from this environment; manipulating f0 contours eliminated effects of the original coda on perceived vowel duration. The effects of the production environment on perceived vowel duration suggest a possible perceptual pathway for the voicing effect on vowel duration.

Cue-shifting between acoustic cues: Evidence for directional asymmetry Publication date: July 2019 Source: Journal of Phonetics, Volume 75 Author(s): Meng Yang, Megha Sundara Abstract Previous research shows that experience with co-varying cues is neither sufficient nor necessary for listeners to integrate them perceptually. Auditory Enhancement theorists explain this by positing that listeners integrate two cues more readily if the cues enhance each other's percept. To isolate the role of enhancement from that of experience, we forced English adult listeners to shift attention between two enhancing cues that they do not use phonemically, pitch and breathiness, by reversing the informativeness of the two cues in a cue weighting experiment. Listeners were able to shift attention from pitch to breathiness and vice versa if the two cues were in an enhancing relation. When this relationship was reversed, listeners could shift attention from pitch to breathiness but not in the opposite direction. Clearly, both the change in informativeness and the enhancing properties of the cues influenced the listeners' re-weighting of these cues. However, the directional asymmetry was not predicted. Moreover, the same asymmetry was observed in two new groups of listeners who have native language experience with either pitch or breathiness. We discuss the consequences of such asymmetric enhancement effects, rising from either processing limitations or articulatory contingencies, for language change.

Formant dynamics of Spanish vocalic sequences in related speakers: A forensic-voice-comparison investigation Publication date: July 2019 Source: Journal of Phonetics, Volume 75 Author(s): Eugenia San Segundo, Junjie Yang Abstract This study investigates the dynamic acoustic properties of 19 vocalic sequences of Standard Peninsular Spanish, showing their potential for forensic voice comparison. Parametric curves (polynomials and discrete cosine transform) were fitted to the formant trajectories of the 19 Spanish vocalic sequences of 54 male speakers, comprising monozygotic (MZ) and dizygotic (DZ) twin pairs, non-twin brothers and unrelated speakers. Using the curve-fitting estimated coefficients as input to a multivariate-kernel-density formula, cross-validated likelihood ratios were calculated to express the probability of obtaining the observed difference between two speech samples under the hypothesis that the samples were produced by the same speaker and under the hypothesis that they were produced by a different speaker. The results show that the best-performing system is one that fuses the 19 vocalic sequences with a geometric-mean fusion method. When challenging the system with related speakers, the results show that MZ twin pairs affect performance but, more importantly, that non-twin sibling pairs can deteriorate performance too. This suggests that more investigations are necessary into a range of similar-sounding speakers beyond MZ twins. Several nurture aspects are highlighted as explanatory factors for the strikingly high similarity of a specific non-twin sibling pair.

Alignment of f0 peak in different pitch accent types affects perception of metrical stress Publication date: May 2019 Source: Journal of Phonetics, Volume 74 Author(s): Katharina Zahner, Sophie Kutscheid, Bettina Braun Abstract In intonation languages, pitch accents are associated with stressed syllables, therefore accentuation is a sufficient cue to the position of metrical stress in perception. This paper investigates how stress perception in German is affected by different pitch accent types (with different f0 alignments). Experiment 1 showed more errors in stress identification when f0 peaks and stressed syllables were not aligned – despite phonological association of pitch accent and stressed syllable. Erroneous responses revealed a response bias towards the syllable with the f0 peak. In a visual-world eye-tracking study (Experiment 2), listeners fixated a stress competitor with initial stress more when the spoken target, which had penultimate stress, was realized with an early-peak accent (f0 peak preceding stressed syllable), compared to a condition with the f0 peak on the stressed syllable. Hence, high-pitched unstressed syllables are temporarily interpreted as stressed – a process directly affecting lexical activation. To investigate whether this stress competitor activation is guided by the frequent co-occurrence of high f0 and lexical stress, Experiment 3 increased the frequency of low-pitched stressed syllables in the immediate input. The effect of intonation on competitor fixations disappeared. Our findings are discussed with respect to a frequency-based mechanism and their implications for the nature of f0 processing.

Language change and linguistic inquiry in a world of multicompetence: Sustained phonetic drift and its implications for behavioral linguistic research Publication date: May 2019 Source: Journal of Phonetics, Volume 74 Author(s): Charles B. Chang Abstract Linguistic studies focusing on monolinguals have often examined individuals with considerable experience using another language. Results of a methodological review suggest that conflating ostensibly 'multicompetent' individuals with monolinguals is still common practice. A year-long longitudinal study of speech production demonstrates why this practice is problematic. Adult native English speakers recently arrived in Korea showed significant changes in their production of English stops and vowels (in terms of voice onset time, fundamental frequency, and formant frequencies) during Korean classes and continued to show altered English production a year later, months after their last Korean class. Consistent with an Incidental Processing Hypothesis (IPH) concerning the processing of ambient linguistic input, some changes persisted even in speakers who reported limited active use of Korean in their daily life. These patterns thus suggest that the linguistic experience obtained in a foreign language environment induces and then prolongs restructuring of the native language, making the multicompetent native speaker in a foreign language environment unrepresentative of a monolingual in a native language environment. Such restructuring supports the view that one's native language continues to evolve in adulthood, highlighting the need for researchers to be explicit about a population under study and to accordingly control (and describe) language background in a study sample.

Intonational structure mediates speech rate normalization in the perception of segmental categories Publication date: May 2019 Source: Journal of Phonetics, Volume 74 Author(s): Jeremy Steffman Abstract The question of if and to what extent listeners' perceptual phonetic categories are sensitive to prosodically driven variability has been a topic of interest in the literature. The present study reports on two experiments which address this question in light of recent research. In Experiment 1, listeners categorized a VOT continuum as /p/ or /b/ in a target syllable (/pɑ/ or /bɑ/). The target was placed in a carrier phrase where the duration and f0 of the pre-target syllable were manipulated. Results suggest listeners are sensitive to intonational structure in their computation of speech rate, interpreting a short syllable with low-rising f0 (created from an L-H% boundary tone in English intonational phonology) as an increase in speech rate. This perceived increase in rate shifts the category boundary of the subsequent target VOT. Experiment 2 showed listeners similarly adjusted categorization of a vowel duration continuum, where vowel duration is a cue to a following obstruent's voicing (categorized as "coat" or "code"). Taken together, these results suggest that listeners are sensitive to intonational structure in their perception of segmental contrasts and use the distribution of tonal targets over a given temporal interval in computing speech rate.

Acoustic correlates of word stress in Papuan Malay Publication date: May 2019 Source: Journal of Phonetics, Volume 74 Author(s): Constantijn Kaland Abstract The current study investigates acoustic correlates of word stress in Papuan Malay. This language is claimed to exhibit word stress, although empirical evidence is lacking. Moreover, related languages have been shown not to have stress, counter to earlier claims. Studies on stress in Austronesian languages have furthermore struggled to separate phrase level intonation phenomena from word level stress. The current study investigates a set of twelve potential acoustic correlates of stress, covering spectral, temporal and amplitudinal aspects of the speech signal. The measurements are taken from spontaneous Papuan Malay narratives. A subsequent comprehensive acoustic analysis was carried out and designed to avoid common pitfalls in the study of word stress. Results indicate that measures of duration, formant displacement and spectral tilt provide consistent evidence for the production of word stress in Papuan Malay.

Twenty-eight years of vowels: Tracking phonetic variation through young to middle age adulthood Publication date: May 2019 Source: Journal of Phonetics, Volume 74 Author(s): Susanne Gahl, R. Harald Baayen Abstract Research on age-related changes in speech has primarily focused on comparing "young" vs. "elderly" adults. Yet, listeners are able to guess talker age more accurately than a binary distinction would imply, suggesting that acoustic characteristics of speech change continually and gradually throughout adulthood. We describe acoustic properties of vowels produced by eleven talkers based on naturalistic speech samples spanning a period of 28 years, from ages 21 to 49. We find that the position of vowels in F1/F2 space shifts towards the periphery with increasing talker age. Based on Generalized Additive Mixed Effects models, we show that this shift is not fully attributable to changes in vowel duration or to segmental context. We discuss the implications of our results for research on aging and speech, and for research in which durational shortening and spectral characteristics of vowels are assumed to reflect a unitary process of phonetic reduction.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Pharmacy And Bioallied Sciences

From the Editorial Desk Gobichettipalayam Jegatheeswaran Anbuselvan, Thuckanickenpalayam Ragunathan Yoithapprabhunath, Thangavelu Arthiie Journal of Pharmacy And Bioallied Sciences 2019 11(6):81-81

Stem cells and their application in dentistry: A review Muraleedhara Bhat, Praveen Shetty, Subramanya Shetty, Faizan A Khan, Shabeeb Rahman, Mallikarjuna Ragher Journal of Pharmacy And Bioallied Sciences 2019 11(6):82-84 The use of the term “stem cells” dates back to the 1800s; however, the application of the same is still not completely understood. Recent advances have indicated the harvesting of postnatal stem cells from sources such as the dental pulp and fat. The pluripotent nature of these cells allows for use in various aspects of treatment and patient care such as organ and tissue transplantation, bony defects repair, distraction osteogenesis, cell therapies, gene therapy, and toxicology testing of new drugs. This article explores the various aspects involved, the current status, and future challenges of stem cell therapy in patient care and management.

Application of finite element model in implant dentistry: A systematic review M Sesha Reddy, Rajasekar Sundram, Hossam Abdelatty Eid Abdemagyd Journal of Pharmacy And Bioallied Sciences 2019 11(6):85-91 FEM was technologically innovated which initially aimed at answering structural analysis difficulties involving Mechanics, Civil and Aeronautical Engineering. FEM basically stands for a numerical model of analyzing stresses as well as distortions in the form of any agreed geometry. There for the shape is discretized into the so-called ‘finite elements’ coupled through nodes. Accuracy of the results is determined by type, planning and total number of elements used for a particular study model. 3-D FE model was designed for in-depth qualitative examination of the relations amongst implant, tooth, periodontal ligament, and bone. Scholarly work equating work reliability, validated with a 3-D modeling suggested that meticulous data can be acquired with respect to stress distribution in bone. Comparative results from 3-D FEA studies showed that 3D FEA, when matched with in-vivo strain gauge measurements were corresponding with clinical outcomes. The aim of this review of literature is to provide an overview to show the application of FEM in (Short) implant dentistry.

Embracing personalized medicine in dentistry Manchala Sesha Reddy, Shishir Ram Shetty, Venkataramana Vannala Journal of Pharmacy And Bioallied Sciences 2019 11(6):92-96 Aim: The aim of this systematic review was to evaluate the number of articles in the area of personalized medicine specific to dentistry. Materials and Methods: Electronic search using three databases was performed using PubMed, Embase, and Scopus search. Results: Results suggest that there is a definite need for more awareness and research pertaining to this specific area. Conclusion: With this background, the authors have written a comprehensive review on applications of personalized medicine in various branches of dentistry.

Periodontal accelerated osteogenic orthodontics technique for rapid orthodontic tooth movement: A systematic review Venkataramana Vannala, Anilkumar Katta, Manchala S Reddy, Shishir R Shetty, Raghavendra M Shetty, Shakeel S Khazi Journal of Pharmacy And Bioallied Sciences 2019 11(6):97-106 Aim: The aim of this review was to analyze the data in the available literature regarding aspects of periodontally accelerated orthodontic therapy such as reduction in treatment time, variation in surgical techniques, and patient satisfaction. Materials and Methods: An internet-based search was performed for the articles published between January 2008 and August 2018 using keywords periodontal accelerated orthodontic tooth movement, corticotomy, bone grafting, tooth movement, and treatment duration. A total of 84 articles were obtained from MEDLINE and Embase search engines, of which 31 articles were eligible to be included for the systematic review. Results: On analysis, it was observed that the earlier studies were predominantly pertaining to case reports. In the later part of the decade considered in this review, it was observed that the emphasis was given to clinical and animal studies. Conclusions: Most of the studies included in the review concluded that there was a significant reduction in orthodontic treatment duration using periodontal accelerated techniques compared to conventional orthodontic treatment.

Immunotherapy in oral cancer Sunil Paramel Mohan, Manoj K Bhaskaran, Ashford L George, Abhilash Thirutheri, Maya Somasundaran, Anegh Pavithran Journal of Pharmacy And Bioallied Sciences 2019 11(6):107-111 Immunotherapy is one of the newer entities which is promising, at least can be very much helpful as an adjuvant therapy. This newer modality of the treatment in the field of cancer treatment may be the fourth pillar supporting surgery, chemotherapy, and radiotherapy. Careful selection of patient is the key for success of immunotherapy, which is based on patient’s immunological contexture. This review aimed to present the fundamental aspects of tumor immunity and immunotherapy, focused on oral squamous cell carcinoma.

Monomer modifications of denture base acrylic resin: A systematic review and meta-analysis R Ajay, K Suma, Seyed Asharaf Ali Journal of Pharmacy And Bioallied Sciences 2019 11(6):112-125 Background: Methyl methacrylate monomer of denture base resins was modified with several monomers to achieve better physico-mechanical properties without compromising the biocompatibility. However, there are no consensuses on the best strategy to achieve best modified monomer. Purpose: To identify and evaluate the differences in the properties between conventional and modified monomers and to verify the influence of several variables on the properties of denture base acrylic resin. Materials and Methods: This study was executed by following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. In-vitro studies that investigated the properties of conventional and modified monomers were selected. Searches were carried out in the Ebscohost, PubMed, Semantic scholar and J-stage databases. The search commenced from the year 1995 and the last search was done till November 2018. A comparison was performed between modified and unmodified monomers. The analyses were carried out using fixed-effect models. Results: The meta-analysis results showed high heterogeneity in all aspects, and higher flexural strength for monomers modified with 20% methacrylic acid. Conclusion: Although the articles included in this meta-analysis showed high heterogeneity and high risk of bias, the in-vitro literature seems to suggest that use of modified monomers could improve the properties of denture base resins. Other variants of monomer modifications and their tested parameters were discussed in this systematic review as well. Dimensional accuracy is an unexplored variable to be evaluated extensively in the future researches.

Platelet-rich plasma and platelet-rich fibrin in periodontal regeneration: A review Sunil Paramel Mohan, Nallusamy Jaishangar, Sandha Devy, Anjhana Narayanan, Deepthi Cherian, Sanupa Sethu Madhavan Journal of Pharmacy And Bioallied Sciences 2019 11(6):126-130 Platelet concentrates (PCs; platelet-rich plasma and platelet-rich fibrin) are autologous bioactive substances that have found varied application in medical and dental fields, particularly in oral and maxillofacial surgery, plastic surgery, and sports medicine. The rationale of these technologies is to extract all the elements from patient’s own blood sample, which could be used to improve healing by promoting tissue regeneration. PCs have evolved a long way since its introduction in 1954. PCs have been used successfully in periodontics and implant dentistry. However, the preparation protocol, processing time, transfer of concentrates, centrifugation temperature, vibration, etc., being not standardized are various factors for the mixed results reported in the literature. This review intends to discuss evolution of PCs, their preparation techniques, and their clinical and technical aspects and applications.

Surface modification techniques for zirconia-based bioceramics: A review Suma Karthigeyan, Arun Jaikumar Ravindran, Ramesh T R Bhat, Madhulika Naidu Nageshwarao, Sree Varun Murugesan, Vignesswary Angamuthu Journal of Pharmacy And Bioallied Sciences 2019 11(6):131-134 Zirconia is gaining interest as a ceramic biomaterial for implant applications due to its biocompatibility and desirable mechanical properties. At present, zirconia-based bioceramics is often seen in the applications of hip replacement and dental implants. This article briefly reviews different surface modification techniques that have been applied to zirconia such as polishing, sandblasting, acid etching, biofunctionalization, coating, laser treatment, and ultraviolet light treatment. The potential of surface modification to make zirconia a successful implant material in the future is highly dependent on the establishment of successful in vitro and in vivo studies. Hence, further effort should be made in order to deepen the understanding of tissue response to implant and tissue regeneration process.

Gingival crevicular fluid: An overview Krishnan Chandragiri Subbarao, Gowri Shankar Nattuthurai, Satheesh Khannaa Sundararajan, Indhu Sujith, Jennifer Joseph, Yasmin Parvin Syedshah Journal of Pharmacy And Bioallied Sciences 2019 11(6):135-139 Gingival crevicular fluid is an inflammatory exudate derived from the periodontal tissues. It is composed of serum and locally generated materials such as tissue breakdown products, inflammatory mediators, and antibodies directed against dental plaque bacteria. It plays a special part in maintaining the structure of junctional epithelium and the antimicrobial defense of periodontium. Some of the suspected periodontal pathogens such as Porphyromonas gingivalis and Treponema denticola produce broad-spectrum neutral proteinases as part of their virulence arsenal. These proteinases may be detected in plaque and gingival crevicular fluid samples of patients with periodontitis. The potential diagnostic importance of gingival fluid was recognized more than six decades ago. The fluid component of gingival crevicular fluid is derived primarily from microvascular (postcapillary venule) leakage. There are number of distinct advantages and challenges of using gingival crevicular fluid as a diagnostic test for periodontal disease.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com