Expression of PTGS2 along with genes regulating VEGF signalling pathway and association with high-risk factors in locally advanced oral squamous cell carcinoma.

BACKGROUND: PTGS2 encodes cyclooxygenase-2 (COX-2), which catalyses the committed step in prostaglandin synthesis. Various in vivo and in vitro data suggest that COX-2 mediates the VEGF signalling pathway. In silico analysis performed in TCGA, PanCancer Atlas for head and neck cancers, demonstrated significant expression and co-expression of PTGS2 and genes that regulate VEGF signalling. This study was designed to elucidate the expression pattern of PTGS2 and genes regulating VEGF signalling in patients with locally advanced oral squamous cell carcinoma (OSCC). METHODOLOGY: Tumour and normal tissue samples were collected from patients with locally advanced OSCC. RNA was isolated from tissue samples, followed by cDNA synthesis. The cDNA was used for gene expression analysis (RT-PCR) using target-specific primers. The results obtained were compared with the in silico gene expression of the target genes in the TCGA datasets. Co-expression analysis was performed to establish an association between PTGS2 and VEGF signalling genes. RESULTS: Tumour and normal tissue samples were collected from 24 OSCC patients. Significant upregulation of PTGS2 expression was observed. Furthermore, VEGFA, KDR, CXCR1 and CXCR2 were significantly upregulated in tumour samples compared with paired normal samples, except for VEGFB, whose expression was not statistically significant. A similar expression pattern was observed in silico, except for CXCR2 which was highly expressed in the normal samples. Co-expression analysis showed a significant positive correlation between PTGS2 and VEGF signalling genes, except for VEGFB which showed a negative correlation. CONCLUSION: PTGS2 and VEGF signalling genes are upregulated in OSCC, which has a profound impact on clinical outcomes.

4-Year Pulp Survival in a Randomized Trial on Direct Pulp Capping.

INTRODUCTION: This study aimed to assess pulp survival in a randomized trial on pulp lavage in adult nonpainful posterior teeth with carious pulp exposure. The treatment included complete caries excavation, direct pulp capping with mineral trioxide aggregate, and immediate restoration with composite resin. METHODS: Fluid was collected from the pulp wound to assess matrix metalloproteinase-9 (MMP-9) and total protein values. Before pulp capping, cavities were randomly (block randomization, n = 48) washed with a physiological saline or a sodium hypochlorite solution (2.5% NaOCl). Treatment outcome was assessed clinically (cold test) and radiographically after at least 1 year and again after at least 3 years. Painful failures were differentiated from nonpainful failures. Pulp survival was estimated using the Kaplan-Meier method including 95% confidence intervals (CIs) up to 1500 days. RESULTS: From the 96 patients originally enrolled, 73 individuals could be followed continuously. The clinical observations indicated a beneficial and sustained effect of pulp lavage with 2.5% NaOCl over a control treatment with physiological saline solution on estimated pulp survival 1500 days postintervention, with 7% (95% CI, 1%-40%) in the saline group versus 55% (95% CI, 30%-100%) in the NaOCl group. High MMP-9/total protein values in pulpal fluid collected from the exposed site indicated early and painful treatment failures yet were not associated with failures that occurred more than 250 days after intervention. CONCLUSIONS: The low 4-year success rates reported here challenge the concept of direct pulp capping in the cases that were included. NaOCl lavage did not only increase the survival of affected pulps substantially but also particularly diminished painful failures (33% in the NaOCl group vs 62% in the saline group). The lack of the predictive value of MMP-9 assessments beyond early treatment failures points to inflammatory states of the pulp tissue under deep caries, which are not related to neutrophil infiltration.

Glucose 6 phosphate dehydrogenase deficiency quantitative test in dried blood spot- a potential marker for adult unknown G6PD deficiency.

Glucose 6 Phosphate Dehydrogenase (G6PD) enzyme activity estimation in a freshly collected blood sample is the most widely used diagnostic method for the diagnosis of G6PD deficiency. The objective is to evaluate the need for newborn screening for G6PD deficiency over post-malarial diagnosis and the feasibility and reliability of using dried blood spots (DBS) as samples for screening. A total of 562 samples were analyzed for G6PD and parallel measurement of G6PD activity by the colorimetric method in whole blood and DBS was carried out in the neonatal subset. Among 466 adults, 27 (5.7%) showed G6PD deficiency, out of whom 22 (81.48%) were diagnosed after the malarial encounter. In the pediatric group, 8 neonates showed G6PD deficiency. G6PD activity estimated from DBS samples agreed with whole blood, with a statistically significant strong positive correlation. Screening of G6PD deficiency at birth to prevent future unwarranted complications, using DBS is feasible.

VARK preference and perception of online versus offline professional development training of medical laboratory technologists.

OBJECTIVE: This study aims to understand the learning preferences and perception of medical laboratory technologists on sudden shift from offline to online training sessions during COVID-19 pandemic. METHODS: Microsoft form containing twenty-four questions was circulated to the twenty-five laboratory technologists after 1 year of online continuous professional development training. VARK questionnaire was circulated to understand the learning style. RESULTS: Provision of recording lectures, significant reduction of performance anxiety, anxiety associated with criticism, and QA sessions emerged as the major positive aspects of a virtual training platform. Analysis of learning preferences revealed that most technologists had a unimodal aural (45%) or kinesthetics (33%) than visual (11%) and reading (11%) learning preference. In bimodal learning preference, AK (44.44%) emerged as the predominant form. Forty percent of the technologists showed trimodal learning pattern with 50% among them showing an ARK pattern while 25% each showing VAK and VRK patterns of learning preferences. CONCLUSION: Medical laboratory technologists adapted well to the sudden shift from offline to online continuous development programs. However, efficient managerial mechanisms to address the major perceived hurdles and designing a multimodal training module to accommodate the learning preferences of our technologists can ensure enthusiastic participation and effective learning among medical laboratory technologists.

Effect of Maleic Acid Root Conditioning on Release of Transforming Growth Factor Beta 1 from Infected Root Canal Dentin.

INTRODUCTION: Chemically released growth factors play a vital role in regenerative endodontics. Transforming growth factor beta 1 (TGF-ß1) is 1 of the most extensively studied bioactive molecules that promotes cell proliferation, differentiation, and chemotaxis. The goal of the current research was to analyze the effect of 7% maleic acid (MA) root conditioning of an infected root canal on the release of TGF-ß1. METHODS: Single-rooted human teeth were decoronated, and the canals were enlarged with a Peeso reamer. The samples were divided into biofilm and nonbiofilm groups. Subsequently, all the samples of both groups were flushed with 10 mL of each irrigant, namely, 1.5% sodium hypochlorite (NaOCl), 7% MA, 17% EDTA, and a combination of 1.5% NaOCl with 17% EDTA or 7% MA, for 10 minutes each. TGF-ß1 was estimated quantitatively using an enzyme-linked immunosorbent assay kit. RESULTS: TGF-ß1 release was lowest among the biofilm samples compared with nonbiofilm among all the groups. MA 7% with 1.5% NaOCl rendered higher amounts of growth factor release in contrast to the combination of 17% EDTA and 1.5% NaOCl in both the biofilm and nonbiofilm groups (P < .048). The nonbiofilm samples treated with 7% MA alone illustrated higher growth factor release compared with 17% EDTA only (P < .006), but there was no significant difference in growth factor release among the biofilm samples treated with 7% MA and 17% EDTA. CONCLUSIONS: Bacterial biofilms modified the release of TGF-ß1. MA 7% was observed to be significantly more efficacious than 17% EDTA in TGF-ß1 growth factor release from radicular dentin.

Unveiling antioxidant and anti-cancer potentials of characterized Annona reticulata leaf extract in 1,2-dimethylhydrazine-induced colorectal cancer in Wistar rats.

BACKGROUND: Colorectal cancer (CC) is the third most common cancer in the world. Annona reticulata (AR) also known as bullock's heart, is a traditional herb. AR leaf extract was initially investigated for its anti-bacterial, anti-inflammatory, anti-malarial, anti-helminthic, anti-stress, and wound healing properties. Only a few in vitro cancer studies have been conducted on AR. Although few studies have linked AR leaf extract to many cancers, comprehensive studies addressing regulation, biological functions, and molecular mechanisms leading to CC pathogenesis are clearly lacking. OBJECTIVES: The present study aimed to explore the antioxidant and anti-cancer potentials of AR leaf extract in CC. MATERIALS AND METHODS: The MTT assay was used to test the anti-proliferative activity of AR leaf extract in vitro on the HCT116 cell line. Qualitative and quantitative phytochemical characterization was carried out using gas chromatography: mass spectrometry (GC-MS). 1,2-dimethylhydrazine (DMH) was used to establish CC model in female Wistar rats. The acute toxicity of AR leaf extract was tested in accordance with OECD guidelines. Aberrant Crypt Foci (ACF) count, organ index, and hematological estimations were used to screen for in vivo anti-cancer potential. The antioxidant activity of colon homogenate was determined. RESULTS: The alcoholic leaf extract (IC50, 0.55 µg/ml) was found to be more potent than the aqueous extract. Using GC-MS, a total of 108 compounds were quantified in the alcoholic leaf extract. The LD 50 value was found to be safe at a dose of 98.11 mg/kg of body weight. AR alcoholic leaf extract significantly (p < 0.05) decreased ACF count and normalized colon length/weight ratio. AR leaf extract increased RBC, hemoglobin and platelets levels. The AR alcoholic leaf extract reduced the DMH-induced tumors and significantly (p < 0.05) increased the activity of endogenous antioxidant enzymes such as catalase, reduced glutathione, superoxide dismutase, and decreased the lipid peroxidase activity. AR leaf extract reduced the inflammation caused by DMH and helped to repair the colon's damaged muscle layers. CONCLUSION: Based on the findings from the present study, it can be concluded that the alcoholic leaf extract of AR has antioxidant and anti-proliferative properties and can aid in the prevention of CC development and dysplasia caused by DMH.

Amino Acid Profile and Lactate Pyruvate Ratio: Potential Adjunct Markers for Differentiating Inborn Errors of Metabolism.

Inborn errors of metabolism (IEM) lead to the physical and mental disability and death of infants, which can be prevented if treated early. Hence it is imperative in diagnosing these disorders at the earliest. The study is planned to differentiate suspected IEM by quantifying the lactate-pyruvate ratio (L/P ratio), Amino acid profiling by HPLC in addition to blood ammonia using the basic methodology in comparison to globally accepted mass spectroscopy technique and specific enzyme activity assays. The objective of this study is not to compete with the existing gold standard methodology rather makes the best of basic diagnostic modalities available. Five cases out of 100 referred cases of IEM suspicion showed the altered pattern of Aminogram correlating with high L/P ratio and Hyperammonemia, which were positive cases, as confirmed by the clinician who referred. Hence study parameters could be used as preliminary protocol to investigate and screen the IEM.

Role of Maternal Serum Human Placental Lactogen in First Trimester Screening.

The most preferred antenatal screening test is first trimester dual test which has a detection rate of 95% for foetal chromosomal anomaly. Maternal serum free ß human chorionic gonadotropin (free ß hCG) and pregnancy associated plasma protein A are used in first trimester dual test along with maternal demographic and foetal sonographic indices to calculate risk for foetal aneuploidy. Human placental lactogen is a placental hormone that is present in maternal serum only during pregnancy and its level rises in relation to the growth of the foetus and placenta. The objectives of this study was to measure and correlate maternal serum hPL with free ß hCG, maternal age, maternal age related risk ratio and calculated risk ratio of first trimester screening. After obtaining permission from the Institutional Ethics Committee, hPL and free ß hCG were measured from the serum of 84 pregnant women aged 20-40 years in 11-13th weeks + 6 days of gestation who underwent dual test during their antenatal check-up. Independent t test, Pearson's correlation, Spearman's correlation, Mann-Whitney U test, ANOVA were used wherever appropriate. A significant positive correlation between maternal serum hPL, maternal age related aneuploidy risk ratio (p value < 0.001) and aneuploidy risk ratio at the time of delivery (p value < 0.001) was observed. Also maternal age was negatively correlated with maternal serum hPL (p value < 0.001) and positively correlated with maternal serum free ß hCG (p value 0.023). A significant negative correlation between maternal serum hPL and free ß hCG (p value < 0.001) was found. To conclude low level of maternal serum hPL in advanced maternal age may reflect decreased functional syncytiotrophoblast mass which may predispose to adverse pregnancy outcome. As chance of baby born with chromosomal anomaly is known to increase with advancing maternal age, hPL may have role in first trimester screening.

Correlation of Red Blood Cell Acetylcholinesterase Enzyme Activity with Various RBC Indices.

Cholinesterases belongs to class hydrolases. There are two types acetylcholinesterase and butyryl cholinesterase. Acetylcholinesterase present in nerve endings and also in the RBC membrane. It helps to maintain the shape and size of RBCs. Any change in shape and size of RBCs may affect the activity of Acetylcholinesterase. Thus this study aimed to estimate RBCs Acetylcholinesterase enzyme activity in various types of anemias and correlate the RBCs Acetylcholinesterase enzyme activity with various hematological indices such as Erythrocyte Sedimentation Rate (ESR), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), Mean Corpuscular Volume (MCV), Red cell Distribution Width (RDW) etc. After obtaining ethical approval from Institutional ethics committee total of 100 samples were collected from Clinical Biochemistry laboratory, Kasturba Medical College, Manipal, Manipal University. 25 were having normal RBC indices, 12 with hemolytic anemia, 26 with microcytic anemia and 26 with macrocytic anemia based on peripheral smear report and RBC indices. Acetylcholinesterase were measured using Ellman's method. RBC acetylcholinesterase activity was significantly increased in microcytic anemia (58.13 ± 5.4) and macrocytic anemia (76.87 ± 6.7) than normal group (37.62 ± 2.71). Also increased RBC acetylcholinesterase was seen in hemolytic anemia (48.11 ± 5.18) but the increase is not statistically significant. RBC acetylcholinesterase correlated negatively with hemoglobin (r = -0.356, p = 0.001) and positively with RDW (r = 0.31, p = 0.003). To conclude RBC acetylcholinesterase activity can be used as one of the potential marker for various types of anemia.

Correlation of Pregnancy Associated Plasma Protein A and Zinc with Calculated Risk Ratio of Dual Test.

INTRODUCTION: First trimester screening by dual test is the most preferred method of antenatal screening. The detection rate of foetal aneuploidy using dual test is 95%. This test uses maternal serum free ß human Chorionic Gonadotropin (free ß hCG) and Pregnancy Associated Plasma Protein A (PAPP-A) along with maternal demographic and foetal sonographic indices to calculate risk for foetal aneuploidy. PAPP-A is a zinc binding metalloproteinase which is also responsible for placental development and foetal growth. So, zinc might also have some role in first trimester screening. No studies are available substantiating the role of zinc in first trimester screening. AIM: To measure and correlate maternal serum zinc with PAPP-A and calculated risk ratio of first trimester screening. MATERIALS AND METHODS: PAPP-A and zinc were measured from the serum of 84 pregnant women aged 20-40 years in 11th-13th weeks+6 days of gestation who underwent dual test during their antenatal check-up. Risk calculation was done using Screening Software for Downs Windows Lab (SSDWL). RESULTS: Maternal serum PAPP-A showed a significant positive correlation with maternal serum zinc (p-value <0.001). Maternal serum PAPP-A and zinc both showed a significant positive correlation with calculated risk ratio (p-value<0.01). CONCLUSION: As PAPP-A is a zinc binding metalloproteinase, zinc showed significant pattern of correlation with aneuploidy risk as shown by PAPP-A. So there could be a possible role for serum zinc in first trimester of screening.