Oral Cavity Fluid as an Alternative Postmortem Matrix: Comparison to Simultaneously Collected Blood and Urine Specimens.

ABSTRACT: In postmortem toxicology analysis, a variety of specimens consisting of fluids and tissues are often collected, each with an intrinsic value. Oral cavity fluid (OCF) is emerging as an alternative matrix in forensic toxicology for contributing to a diagnosis in postmortem cases; especially when blood is limited or not available. The aim of this study was to assess the analytical results obtained from OCF and compare them with blood, urine, and other traditional matrices collected from the same postmortem subjects. Of the 62 decedents studied (including 1 stillborn, 1 charred, and 3 decomposed subjects), 56 had quantifiable drugs and metabolites data in the OCF, blood, and urine. Notable findings were benzoylecgonine (24 cases), ethyl sulfate (23 cases), acetaminophen (21 cases), morphine (21 cases), naloxone (21 cases), gabapentin (20 cases), fentanyl (17 cases), and 6-acetylmorphine (15 cases), which were detected more frequently in OCF than in blood (heart, femoral, or body cavity) or urine. This study suggests that OCF is a suitable matrix for detecting and quantifying analytes in postmortem subjects compared with traditional matrices, particularly when other matrices are limited or difficult to collect because of body condition or putrefaction.

Synthesis and in vitro antibacterial behavior of curcumin-conjugated gold nanoparticles.

Owing to the rise in multidrug-resistant bacterial diseases and the dwindling supply of newer antibiotics, it is crucial to discover newer compounds or modify current compounds for more effective antimicrobial therapies. According to reports, more than 80% of bacterial infections have been linked to bacterial biofilms. In addition to having antimicrobial properties, the hydrophobic polyphenol curcumin (Cur) also inhibits quorum sensing. The application of curcumin was constrained by its weak aqueous solubility and quick degradation. Over the past years, nanotechnology-based biomaterials with multi-functional characteristics have been engineered with high interest. The present study focused on the development of nano-biomaterials with excellent testifiers for bacterial infection in vitro. In this study, water dispersibility and stability of curcumin were improved through conjugation with gold nanoparticles. The successful synthesis of curcumin-conjugated gold nanoparticles (Cur-AuNPs) was confirmed using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and UV-vis absorbance spectroscopy. Transmission electron microscopy (TEM) revealed an average particle size of about 10-13 nm. The antibacterial characteristics in terms of the minimum inhibitory concentration (MIC) of Cur-AuNP treatments were found to be lowest than those with AuNPs and Cur treatments. The quantitative analysis revealed the superior antibacterial characteristics of Cur-AuNP-treated bacterial cells compared to the untreated samples. In addition, curcumin-conjugated AuNPs, produced more reactive oxygen species and increased the membrane permeability. Besides, the biocompatibility of Cur-AuNPs was also assessed quantitatively and qualitatively. Statistical analyses revealed the augmented MG-63 cell proliferation in Cur-AuNPs compared to those with Cur and AuNPs treatments. Overall, Cur-AuNPs exhibited enhanced antibacterial, and antibiofilm characteristics and cytocompatibility.

Fatal Intoxication by the Novel Cathinone 4-Fluoro-3-methyl-α-PVP.

A 30 year old non-hispanic white male was found unresponsive at his workplace and admitted to the hospital in cardiac arrest. He was pronounced deceased shortly after arrival. At autopsy the pathologist noted a 176 pound, well-nourished, atraumatic, adult male with significant bilateral frothy pulmonary edema (right lung 930 g and left lung 1,130 g), cardiomegaly (430 g), dilated ventricles and slight cerebral edema. Upon completion of the systematic toxicological analysis scope for the Franklin County Coroner's Office Toxicology Laboratory, no known drugs were found. Further review of the gas chromatography--mass spectrometry (GC--MS) full-scan library summary reports showed an unknown peak in both the blood and urine solid phase extracts. An analogue of α-pyrrolidinovalerophenone (α-PVP) was identified, and a GC--MS selected ion monitoring method was developed to identify and quantitate the presence of 4-fluoro-3-methyl-α-PVP. This method quantified the drug at 26 ng/mL in gray top femoral blood, 30 ng/mL in purple top heart blood and 20 ng/mL in red top vitreous humor. Qualitative presence was also observed in the urine but was not detected in the liver. The decedent's cause of death was determined to be due to fluoro-methyl-PVP toxicity and the manner was ruled to be accidental. Investigational follow-up interviews corroborated drug use by the deceased with a preference of research chemicals and synthetic cannabinoids via the internet. No published literature is available currently, and to the author's knowledge this is the first incident of a fatal death solely attributed to this substituted cathinone.

Synthesis of biocompatible, BSA capped fluorescent CaCO3 pre-nucleation nanoclusters for cell imaging applications.

Novel, photostable, multicolour fluorescent, highly biocompatible, water soluble, BSA capped pre-nucleation CaCO3 nanoclusters (FCPN) (∼1.3 nm) are developed using a facile biomineralization process. M. oleifera leaf extract and BSA protein are used as sources of ascorbic acid and capping agent, respectively. The developed FCPN shows fluorescence in the blue, green, and yellow/red region with an average life time of 1.05, 6.23 and 30.60 ns, respectively. The MALDI-MS measurements reveal that these nanoclusters are 16, 50, 73, 222 and 936 molecules big. These FCPN, when incubated (up to 7 days) with MG-63 cells, demonstrate an increase in cell viability percentage with time period as compared to their control samples. Furthermore, these incubated cells were investigated using confocal microscopy to estimate the FCPN diffusion penetration depth using CTCF analysis. It has been observed that blue and green emitting FCPN penetrated 6 µm, whereas red emitting FCPN traversed only 4 µm. The relative quantum yield (Rhodamine 6G = 0.92) of FCPN for green emission was found to be 0.0175 in water. The prepared nanoclusters displayed four months shelf-life. These FCPN were prepared using an environmentally benign, inexpensive, green synthetic route without using toxic reducing agents. Furthermore, the current report discusses the detailed results, obtained from X-ray photoelectron spectroscopy, MALDI-MS, Fourier transform infrared spectroscopy, UV-visible, fluorescence spectroscopy, lifetime measurements, electron microscopy, fluorescence microscopy and confocal microscopy.

Flagellar targeting of an arginine kinase requires a conserved lipidated protein intraflagellar transport (LIFT) pathway in Trypanosoma brucei.

Both intraflagellar transport (IFT) and lipidated protein intraflagellar transport (LIFT) pathways are essential for cilia/flagella biogenesis, motility, and sensory functions. In the LIFT pathway, lipidated cargoes are transported into the cilia through the coordinated actions of cargo carrier proteins such as Unc119 or PDE6δ, as well as small GTPases Arl13b and Arl3 in the cilium. Our previous studies have revealed a single Arl13b ortholog in the evolutionarily divergent Trypanosoma brucei, the causative agent of African sleeping sickness. TbArl13 catalyzes two TbArl3 homologs, TbArl3A and TbArl3C, suggesting the presence of a conserved LIFT pathway in these protozoan parasites. Only a single homolog to the cargo carrier protein Unc119 has been identified in T. brucei genome, but its function in lipidated protein transport has not been characterized. In this study, we exploited the proximity-based biotinylation approach to identify binding partners of TbUnc119. We showed that TbUnc119 binds to a flagellar arginine kinase TbAK3 in a myristoylation-dependent manner and is responsible for its targeting to and enrichment in the flagellum. Interestingly, only TbArl3A, but not TbArl3C interacted with TbUnc119 in a GTP-dependent manner, suggesting functional specialization of Arl3-GTPases in T. brucei These results establish the function of TbUnc119 as a myristoylated cargo carrier and support the presence of a conserved LIFT pathway in T. brucei.

Sandalwood-derived carbon quantum dots as bioimaging tools to investigate the toxicological effects of malachite green in model organisms.

Malachite green is an N-methylated diaminophenylmethane dye that has generated much concern over its suggestive carcinogenic nature. After its excessive use in aquaculture industry as an effective ectoparasitide, much debate was raised over its toxicological effects leading to scientific studies conducted on animal models. Even after several bans, malachite green is still easily available in many parts of the world and unscrupulously even used to give green vegetables a fresher look. This study aims to address this concern by systematically studying the toxicological effects of malachite green through bioimaging in plant and animal cell and tissue. Sandalwood-derived carbon quantum dots have been used as a bioimaging tool since they are non-cytotoxic and show excellent fluorescence properties. Onion tissues demonstrate the translocation of the dye inside cells having high affinity for the nuclei and cell walls. Toxicological effects on the growth of Vigna radiata (mung beans) have been studied methodically. Bioimaging of the transverse cross-section of the dye-treated plant root shows a significant difference from the control. In animal cells, dose-dependent decrease in cell viability of MG-63 cells was observed with MG. CQD showed good fluorescence in both cytoplasm and nucleus of MG63 cells. In addition, CQDs were employed as a great tool for bioimaging of the histopathologically adverse effects of MG in Golden hamster animal model. This study showed CQDs could be used as an alternative non-site specific fluorescent probe for cell and tissue imaging for better visualization of cell and tissue architectural changes.

Identification of microRNA-target modules from rice variety Pusa Basmati-1 under high temperature and salt stress.

High temperature and salinity stress are major factors limiting the growth and productivity of rice crop on a global scale. It is therefore an essential prerequisite to understand the molecular genetic regulation of plant responses to dual stresses. MicroRNAs (miRs) are recognized as key controllers of gene expression which act mainly at the post-transcriptional level to regulate various aspects of plant development. The present study attempts to investigate the miR circuits that are modulated in response to high temperature and salinity stress in rice. To gain insights into the pathway, preliminary miR profiles were generated using the next-generation sequencing (NGS) datasets. The identified molecules were filtered on the basis of fold differential regulation under high temperature, and time kinetics of their expression under the two individual stresses was followed to capture the regulatory windows. The analysis revealed the involvement of common miR regulatory nodes in response to two different abiotic stresses, thereby broadening our perspective about the stress-mediated regulatory mechanisms operative in rice.

Prevalence of extended spectrum beta-lactamase producing uropathogens and their antibiotic resistance profile in patients visiting a tertiary care hospital in central India: Implications on empiric therapy.

CONTEXT: Antimicrobial resistance showed by different uropathogens is one of the barricades that might hinder a successful treatment. Detection of extended spectrum beta-lactamase (ESBL) production among uropathogens is an important marker of endemicity. AIMS: The present prospective study was done to identify the trends of uropathogens, to find the prevalence of ESBL isolates and to study the antibiotic resistance profile of the ESBL and non-ESBL uropathogenic isolates. MATERIALS AND METHODS: This study was conducted in the Department of Microbiology of a teaching tertiary care hospital from July 2013 to September 2013. All the uropathogenic isolates were identified up to species level by conventional methods. The prevalence of potential ESBL producers was explored. Antibiotic resistance test of the urinary isolates was done by disc-diffusion method and the results were interpreted according to Clinical Laboratory Standards Institute-2013 guidelines. RESULTS: A total of 670 urine samples from male and female patients visiting the outpatient department (OPD) and inpatient department (IPD) of our hospital were collected. A significantly higher number of IPD and OPD males (55.1% and 55.5%) were found to be culture positive. Escherichia coli (55.3%) was the most frequently isolated uropathogen followed by Klebsiella pneumoniae (23%). However, strains of Escherichia coli (41.6%) were the highest ESBL producing isolates followed by Pseudomonas aeruginosa (36.1%). ESBL producing isolates were found to be multidrug-resistant when compared to non-ESBL producers. However, excessive drug-resistance among non-ESBL producing isolates can't be ignored. CONCLUSION: Our study confirms a global trend toward increased resistance to beta-lactam antibiotics. We emphasize on the formulation of antibiotic policy for a particular geographical area.

Boswellic acid inhibits inflammatory angiogenesis in a murine sponge model.

The aim of the present study was to investigate the effects of boswellic acid (BA) on key components of inflammatory angiogenesis in the murine cannulated sponge implant angiogenesis model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and BA (12.5 or 25mg/kg/day) was given through installed cannulas for nine days. The implants collected at day 9 post-implantation were processed for the assessment of hemoglobin (Hb). Relevant levels of inflammatory, angiogenic and fibrogenic cytokines were also determined. BA treatment resulted in significant decrease in sponge vascularization (Hb content) and in vascular endothelial growth factor (VEGF) and transforming growth factor (TGF-ß1) at both doses. Further, BA decreased expression of VEGF and CD31 and reduced % microvessel density (MVD) in sponge implants. A regulatory function of BA on multiple parameters of the main components of inflammatory angiogenesis has been revealed giving an insight into the potential therapeutic use underlying the actions of BA.

Cytotoxic and antitumor effects of brucine on Ehrlich ascites tumor and human cancer cell line.

AIMS: Brucine (BRU), a natural plant alkaloid is reported to possess cytotoxic and antiproliferative activities. In this study we aimed to investigate its in vitro and in vivo antitumor and antiangiogenic effects. MAIN METHODS: Cell proliferation and viability was assessed using microculture tetrazolium tests (MTT). As predictive markers we determined intracellular levels of vascular endothelial growth factor (VEGF), Interleukin-12 (IL-12), tumor necrosis factor (TNF-α), caspase-3, -8 and -9 by ELISA and enzymatic activity assays. In addition, anti-VEGF neutralization effect was evaluated to assess whether it could result in augmented anticancer efficacy than the single agent. Antitumor activity was evaluated against Ehrlich ascites and solid tumor models. 15×10(6) EAC cells were implanted intraperitoneally (i.p., ascites tumor) and subcutaneous (s.c., solid tumor) in Swiss albino mice. Mice with established tumors received brucine i.p. at 12.5, 25, and 50mg/kg for 14days in ascites tumor and 50mg/kg in solid tumor for 30days. Tumor volume, cell viability, angiogenic, anti-angiogenic, anti-inflammatory factors and antioxidant parameters were determined. Immunohistochemistry analysis for VEGF and CD-31 was also performed. KEY FINDINGS: BRU produced time and dose-dependent inhibition of MCF-7 in vitro and EAC tumors in vivo. The anti-angiogenic effects were accompanied with decreased VEGF and TNF-α and increased IL-12 expression. BRU reduced peritoneal angiogenesis and microvessel density in vivo. CONCLUSION: Our findings suggest that BRU possesses antitumor and anti-angiogenic activities in vitro and in vivo. The above results showed that BRU can be used as a potential anticancer agent as an antimetastatic and anti-angiogenic agent.

Antitumor properties of Boswellic acid against Ehrlich ascites cells bearing mouse.

Boswellic acid (BA), a triterpene, isolated from Boswellia serrata (Burseraceae) has been found to possess potent anti-inflammatory and anti-cancer activity. The present study aimed at exploring the possible role of BA on ascites and solid Ehrlich tumor. Ascitic tumor development was evaluated 14 d after tumor implantation by quantification of the ascitic fluid volume whereas solid tumor was evaluated after 30 d tumor implantation by H&E and IHC. The i.p. administration of BA significantly inhibited ascitic and solid Ehrlich tumor model. This inhibition was observed with reduced ascitic volume, solid tumor volume and body weight when compared to control mice. The treatments also increased the survival of tumor-bearing mice. VEGF and TNF- α levels were decreased, whereas the IL-12 levels were increased with BA treatment at 25mg/kg. Further, results on decrease in the peritoneal angiogenesis and microvessel density showed the anti-angiogenic potential. Microscopic examination of tumors revealed that in BA-treated groups the expression of Bax and caspase 3 increased, suggesting drug induced tumor cell apoptosis through activating the pro-apoptotic bcl-2 family and caspase-3. The present study sheds light on the potent antitumor property of the boswellic acid and can be extended further to develop therapeutic protocols for treatment of cancer.