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Background: Health-care workers (HCWs) can sustain needlestick injuries (NSIs) during patient care activities and are at a greater risk of exposure to blood-borne infectious agents such as HBV, HCV, and HIV, among which only HBV has an effective vaccine. Unfortunately, both the voluntary reporting rates of NSIs and HBV vaccination coverage among HCWs have been generally low. This study aims to analyze the trends of voluntary reporting of NSIs and the HBV vaccination status among the HCWs of our tertiary health care center., Materials and Methods: A retrospective study was conducted between January 2016 and July 2018 at a tertiary health care center in Puducherry. A standard pro forma was filled by all HCWs reporting NSIs. HIV, hepatitis B virus surface antigen, and hepatitis C virus were tested using rapid card tests, and anti-hepatitis B surface titers were tested using enzyme-linked immunosorbent assay according to the manufacturer's instructions., Results: A total of 83 incidents of NSIs were reported. Nursing staffs reported the maximum number of incidents followed by students. About 42 incidents of NSI were reported in the year 2018 compared to 26 and 15 incidents reported in 2017 and 2016, respectively. Only 37 HCWs had taken the complete course of HBV vaccine., Conclusion: This study shows an increase in the voluntary reporting of NSIs by the HCWs, but the HBV vaccination status among them was found to be low., Competing Interests: There are no conflicts of interest., (Copyright: © 2019 Journal of Laboratory Physicians.)

Objectives: Pathogenesis of gallstone includes bile stasis due to defect in the gallbladder muscle contraction. Our aim of the study is to find out the role of 99mTc-HIDA scan in assessment of gallbladder dyskinesia in cholelithiasis patients before laparoscopic cholecystectomy and compare the gallbladder dyskinesia with various parameters like symptoms of patients, diabetic status of patients, gallstones size and number, and cholecystitis features in histopathology report after surgery., Material and Method: This is a prospective observational study conducted at our hospital for three years. Totally 40 patients with gallstone were subjected to 99mTc-HIDA scan, to assess the ejection fraction of gallbladder. For all these patients detailed clinical history, presence of comorbid illness like diabetics, and symptomatology were elicited. For all patients, ultrasonogram of abdomen was done to assess number and size of stones. All parameters were tabulated and correlated., Result: While comparing 99mTc-HIDA scan findings with symptoms of patients, 21.2% were asymptomatic and 78.8% symptomatic patients who had ejection fraction less than 80%. All patients in EF >80% group were symptomatic only. It is not statistically significant. On comparing 99mTc-HIDA scan findings with diabetic status of the patients, 42.4% of diabetic and 57.6 % of nondiabetic patients had ejection fraction less than 80%. It is not statistically significant (0.681). While comparing 99mTc-HIDA scan findings with size of the gallstone in ultrasound, 63.6% patients with size less than 1cm and 36.4% with size more than 1cm had ejection fraction < 80%. It is statistically significant (0.048). On comparing 99mTc-HIDA scan findings with number of stones in ultrasound, 18.2% single gallstone patients and 81.8% multiple gallstone patients had EF less than 80% which is statistically significant (0.001). While comparing the 99mTc-HIDA scan findings with histopathology report after laparoscopic cholecystectomy, 21.2% non-cholecystitis patients and 78.8% cholecystitis patients had EF less than 80%, which is statistically (0.017) significant., Conclusion: 99mTc-HIDA scan can be an accurate method to diagnose the gallbladder dyskinesia. Gallbladder dyskinesia in 99mTc-HIDA scan can be used to predict large size stones and multiple stones before surgery. The sensitivity can be improved by 99mTc-HIDA scan in diagnosing cholecystitis patients.

[This corrects the article DOI: 10.1155/2016/3059434.].

Film rehydration method is commonly used for membrane protein (MP) reconstitution into block copolymer (BCP), but the lack of control in the rehydration step formed a heterogeneous population of proteopolymersomes that interferes with the characterization and performance of devices incorporating them. To improve the self-assembly of polymersomes with simultaneous MP reconstitution, the study reported herein aimed to understand the effects of different variants of the rehydration procedure on the MP reconstitution into BCP membranes. The model MP used in this study was AquaporinZ (AqpZ), an α-helical MP that has been shown to have a high permeation rate exclusive to water molecules. Comparing four rehydration methods differing in the hydration time (i.e., brief wetting or full hydration) and medium (i.e., in buffer or AqpZ stock solution), prehydration with buffer prior to adding AqpZ was found to be most desirable and reproducible reconstitution method because it gave rise to the highest proportion of well-formed vesicles with intact AqpZ functionality as evidenced by the transmission electron microscopy images, dynamic light scattering, and stopped-flow analyses. The mechanisms by which effective AqpZ reconstitution takes place were also investigated and discussed. Small-angle X-ray scattering analysis shows that hydrating the initially dry multilamellar BCP films allows the separation of lamellae. This is anticipated to increase the membrane fluidity that facilitates a fast and spontaneous integration of AqpZ as the detergent concentration is considerably lowered below its critical micelle concentration. Dilution of detergent can result in precipitation of proteins in the absence of well-fluidized membranes for protein integration that underscores the importance of membrane fluidity in MP reconstitution.

Objectives. In our study we are aiming to analyse the learning curves in our surgical trainees by using two standard methods of intracorporeal knot tying. Material and Method. Two randomized groups of trainees are trained with two different intracorporeal knot tying techniques (loop and winding) by single surgeon for eight sessions. In each session participants were allowed to make as many numbers of knots in thirty minutes. The duration for each set of knots and the number of knots for each session were calculated. At the end each session, participants were asked about their frustration level, difficulty in making knot, and dexterity. Results. In winding method the number of knots tied was increasing significantly in each session with less frustration and less difficulty level. Discussion. The suturing and knotting skill improved in every session in both groups. But group B (winding method) trainees made significantly higher number of knots and they took less time for each set of knots than group A (loop method). Although both knotting methods are standard methods, the learning curve is better in loop method. Conclusion. The winding method of knotting is simpler and easier to perform, especially for the surgeons who have limited laparoscopic experience.

Introduction: Despite the advances in the treatment modalities offered for oral squamous cell carcinoma. The recurrence rate of it still remains quite high. Early detection of recurrence will improve the outcome and the survival of the patient. Osteopontin, a transformation-related phosphorylated protein in epithelial cells has been closely related with tumourigenesis. This study was undertaken to explore the potential of OPN as a tumour marker of recurrence in OSCC., Aim: To analyse the expression of Osteopontin (OPN) in Oral Squamous Cell Carcinoma (OSCC), patient matched tumour free surgical margins and normal oral mucosa and to correlate with local & loco regional recurrence., Materials and Methods: Twenty cases each of formalin fixed paraffin embedded blocks of histopathologically diagnosed cases of OSCC, patient matched tumour free surgical margins and normal oral mucosal tissues were obtained from the archives of the Oral Pathology & Microbiology Department, Faculty of Dental Sciences, SRU and Govt. Arignar Anna Memorial Cancer Hospital, Kancheepuram. Immunohistochemical analysis was performed with an antibody to Osteopontin protein. Patients with secondary tumours and those treated with chemotherapy and radiotherapy were excluded from this study., Results: The expression of OPN was elevated in 95% of tumours & 55% of histologically tumour free margin samples. There was negative OPN expression in normal mucosal samples. The result of the study was statistically analysed using Pearson chi-square test and was found to be statistically significant., Conclusion: OPN can be used as a diagnostic marker in Oral Squamous Cell Carcinoma. In the tumour free surgical margins, elevated levels of OPN may predict a significantly increased risk of recurrence.

Osteopontin (OPN), a matrix extracellular glyco-phosphoprotein is found in various tissues such as epithelium lined tissues, kidney, bone and teeth .It is also detected in all body fluids including blood and breast milk. OPN plays role in a number of physiological and pathologic events such as cell adhesion, migration and cell survival, angiogenesis, apoptosis, inflammation and wound healing. This review summarizes the current data of the biological activities of OPN in the development of tumour, its progression and metastasis.

Cold atmospheric plasma (CAP), an ambient temperature ionized gas, is gaining extensive interest as a promising addition to anti-tumor therapy primarily due to the ability to generate and control delivery of electrons, ions, excited molecules, UV photons, and reactive species such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) to a specific site. The heterogeneous composition of CAP offers the opportunity to mediate several signaling pathways that regulate tumor cells. Consequently, the array of CAP generated products has limited the identification of the mechanisms of action on tumor cells. The aim of this work is to assess the cell death response of human myeloid leukemia cells by remote exposure to CAP generated RNS by utilizing a novel resistive barrier discharge system that primarily produces RNS. The effect of variable treatments of CAP generated RNS was tested in THP-1 cell (human monocytic leukemia cell line), a model for hematological malignancy. The number of viable cells was evaluated with erythrosine-B staining, while apoptosis and necrosis was assessed by endonuclease cleavage observed by agarose gel electrophoresis and detection of cells with the exclusionary dye propidium iodide and fluorescently labeled annexin-V by flow cytometry and fluorescent microscopy. Our observations indicate that treatment dosage levels of 45 s of exposure to CAP emitted RNS-induced apoptotic cell death and for higher dosage conditions of ≥50 s of exposure to CAP induced necrosis. Overall the results suggest that CAP emitted RNS play a significant role in the anti-tumor potential of CAP., (© 2013 Wiley Periodicals, Inc.)

Introduction: Chordomas are rare, slow growing, locally destructive bone tumours arising from the notochord., Presentation of Case: Presenting a case of a 65 year old man, who presented with complaints of swelling on the right lower back for 1 year associated with pain. On, physical examination, a swelling measuring 5cm×4cm was noted in the lower back with posterior wall indentation on per rectal examination. MRI revealed a mass lesion involving the sacrum (s3-s4) and coccyx. FNAC showed features of a chroma. At surgery, we excised a mass from the retrorectal space and biopsy proved it to be a chondroid chordoma, a variant of chordoma., Discussion: Chordomas are solid malignant tumours that arise from vestiges of the foetal notochord. Common locations are the clivus and the sacrococcygeus region. Annual incidence of these tumours is 1 in one million. MRI is the imaging modality of choice. Prognosis improves based on the age, resected margins and postoperative treatment., Conclusion: Here, we shall discuss the literature, variants, treatment and prognosis of this rare tumour., (Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The conservation of sleep across all animal species suggests that sleep serves a vital function. We here report that sleep has a critical function in ensuring metabolic homeostasis. Using real-time assessments of tetramethylammonium diffusion and two-photon imaging in live mice, we show that natural sleep or anesthesia are associated with a 60% increase in the interstitial space, resulting in a striking increase in convective exchange of cerebrospinal fluid with interstitial fluid. In turn, convective fluxes of interstitial fluid increased the rate of β-amyloid clearance during sleep. Thus, the restorative function of sleep may be a consequence of the enhanced removal of potentially neurotoxic waste products that accumulate in the awake central nervous system.

Background: Neurodegenerative diseases such as Alzheimer's are associated with the aggregation of endogenous peptides and proteins that contribute to neuronal dysfunction and loss. The glymphatic system, a brain-wide perivascular pathway along which cerebrospinal fluid (CSF) and interstitial fluid (ISF) rapidly exchange, has recently been identified as a key contributor to the clearance of interstitial solutes from the brain, including amyloid β. These findings suggest that measuring changes in glymphatic pathway function may be an important prognostic for evaluating neurodegenerative disease susceptibility or progression. However, no clinically acceptable approach to evaluate glymphatic pathway function in humans has yet been developed., Methods: Time-sequenced ex vivo fluorescence imaging of coronal rat and mouse brain slices was performed at 30-180 min following intrathecal infusion of CSF tracer (Texas Red- dextran-3, MW 3 kD; FITC- dextran-500, MW 500 kD) into the cisterna magna or lumbar spine. Tracer influx into different brain regions (cortex, white matter, subcortical structures, and hippocampus) in rat was quantified to map the movement of CSF tracer following infusion along both routes, and to determine whether glymphatic pathway function could be evaluated after lumbar intrathecal infusion., Results: Following lumbar intrathecal infusions, small molecular weight TR-d3 entered the brain along perivascular pathways and exchanged broadly with the brain ISF, consistent with the initial characterization of the glymphatic pathway in mice. Large molecular weight FITC-d500 remained confined to the perivascular spaces. Lumbar intrathecal infusions exhibited a reduced and delayed peak parenchymal fluorescence intensity compared to intracisternal infusions., Conclusion: Lumbar intrathecal contrast delivery is a clinically useful approach that could be used in conjunction with dynamic contrast enhanced MRI nuclear imaging to assess glymphatic pathway function in humans.

The aim of this study was to assess the open pond and groundwater quality of Tiruchirapalli city of Tamil Nadu, India. The groundwater quality viz., pH, electrical conductivity, total hardness, calcium ion, magnesium ion, chloride, carbonate, bicarbonate, inorganic nitrate, nitrite, phosphate, ammonia and reactive silicate were analysed with respect to various seasons and recorded in the range of 7.1 to 8.1, 97.67 to 533.67 mhos cm(-1), 7.07 to 186 mg l(-1), 4.67 and 112.0 mg l(-1), 2.40 to 92.80 mg l(-1), 15.23 to 661.73 mg l(-1), 60 to 480 mg l(-1), 22.7 to 544.9 mg l(-1), 15.33 to 68.00 mg l(-1), 0.001 to 0.480 mg l(-1), 0.01 to 0.42 mg l(-1), 0.02 to 0.75 mg l(-1) and 1.1 to 2.96 mg l(-1) respectively. The present findings concluded that the quality of ground waters can be considered suitable for human consumption. But the pond water available in and around Tiruchirappalli city was not fit for human usage, agricultural or industrial purposes.

In this work an atmospheric pressure non-thermal resistive barrier (RB) plasma jet was constructed, characterized and was applied for biomedical applications. The RB plasma source can operate in both DC (battery) as well as in standard 60/50 Hz low frequency AC excitation, and it functions effectively in both direct and indirect plasma exposure configurations. The characteristics of the RB plasma jet such as electrical properties, plasma gas temperature and nitric oxides concentration were determined using voltage-current characterization, optical emission spectroscopy and gas analyzer diagnostic techniques. Plasma discharge power of 26.33 W was calculated from voltage-current characterization. An optical emission spectroscopy was applied and the gas temperature which is equivalent to the nitrogen rotational (Trot) temperatures was measured. The concentrations of the reactive oxygen species at different spatial distances from the tip of the plasma jet were measured and the ppm concentration of NO is at the preferred level for a wide range of standard biomedical treatment applications. The ppm values of nitric oxides after the cooling unit are observed to be of the same order of magnitude as compared to plasma jet. The portable RB plasma source was tested to be very effective for decontamination and disinfection of a wide range of foodborne and opportunistic nosocomial pathogens such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus and the preliminary results are presented. The effects of indirect exposure of the portable RBP source on monocytic leukemia cancer cells (THP-1) were also tested and the results demonstrate that a preference for apoptosis in plasma treated THP-1 cells under particular plasma parameters and dosage levels.

In the brain, a paravascular space exists between vascular cells and astroglial end-foot processes, creating a continuous sheath surrounding blood vessels. Using in vivo two-photon imaging we demonstrate that the paravascular circulation facilitates selective transport of small lipophilic molecules, rapid interstitial fluid movement and widespread glial calcium signaling. Depressurizing the paravascular system leads to unselective lipid diffusion, intracellular lipid accumulation and pathological signaling in astrocytes. As the central nervous system is devoid of lymphatic vessels, the paravascular space may serve as a lymphatic equivalent that represents a separate highway for the transport of lipids and signaling molecules.

This research investigated the modulation of cell death through exposure of non-thermal resistive barrier based indirect air plasma on monocytic leukemia cancer cells (THP-1). Specifically, we explored cell death through apoptosis and necrosis, since generally apoptotic cell death has a limited inflammatory response as compared to necrosis. We have demonstrated a preference for apoptosis in plasma treated THP-1 cells, under specific plasma characteristics and dosage levels, using fluorescent dyes conjugated with annexin V followed by identification of the cells through fluorescent microscopy and flowcytometry diagnostics. At much higher plasma dosages, the necrotic morphologies in the THP-1 cells were observed. The presented outcomes in the death morphologies of plasma treated THP-1 cells signify the need for further investigation on the cellular mechanisms induced by the indirect plasma exposure. The results obtained from this research indicate the significant potential for the use of our portable non-thermal resistive barrier based indirect plasma treatment method as an inexpensive and less invasive method for treating leukemia and other cancerous lesions.

The purpose of this study was to determine the effects of plasma treatment on bacteria in liquid phases. We predict that the plasma gas can penetrate the liquid culture media and plasma treatment will efficiently kill the bacteria at unique time and distance parameters. It is also hypothesized that less stringent plasma treatment will negatively affect the growth rate of some species of bacteria and possibly their pathogenicity. The bacteria were exposed to hot and cold plasma at various time lengths and distance parameters. Our results indicated that 2 minutes of hot plasma treatment with the plasma torch 5 cm away from the liquid culture is effective in killing/sterilizing cultures of S. aureus, S. pyogenes, Salmonella spp, N. meningitidis, and E. coli. Five minutes of cold plasma with the probe immersed 1-2 cm inside the liquid culture were needed to kill the bacteria. The portable nonthermal plasma system can be used for infection treatment and wound healing applications affected by the microbes studied in this research.

This research study examined the effect of non-thermal portable atmospheric air plasma system on leukemia cancer cells. Acute monocytic leukemia cells (THP-1) were exposed to atmospheric pressure non-thermal plasma. To assess death caused by plasma exposure, cells were subjected to trypan blue exclusion assays and a kill-curve and assessment of death overtime were compiled using data from the assays. In addition to this, DNA was harvested from treated and untreated samples to determine if apoptotic ladders were present. Results have indicated that non-thermal plasma can cause cell death in THP-1 cells overtime, and the death that occurs corresponds directly to the amount of time that the cells were exposed to ionized plasma. Preliminary fluorescent imaging of the treated cells revealed that higher treatment doses are not only more likely to induce cellular death but are likely to induce necrotic death, while lower treatment doses that are capable of inducing death may induce apoptotic or programmed cellular death. Ideally the results obtained from these experiments will allow for further investigation of the effects of ionized non-thermal plasma on melanoma cell lines and will lead to an inexpensive method for treating early stage skin cancer and cancerous lesions.

Objective: Activated protein C (APC) is neuroprotective in stroke models and promotes postischemic neovascularization and neurogenesis. We used a controlled cortical impact (CCI) in mice to determine the effects of APC on neuroprotection and angiogenesis and neurogenesis after traumatic brain injury (TBI)., Methods: Mice were given (1) single-dose APC (0.8 mg/kg intraperitoneally) 15 minutes after injury, (2) multidose APC (0.8 mg/kg intraperitoneally) 15 minutes and 6 to 48 hours after injury, or (3) vehicle. We then assessed the effects of APC on posttraumatic motor function with the rotarod and wire grip and beam balance tasks, and we determined the lesion volumes and studied the formation of new blood vessels and markers of neurogenesis., Results: Mice treated with single-dose or multidose APC, compared with vehicle, showed significantly improved motor function on all tests. In the single-dose and multidose APC treatment groups, at 7 days after treatment, lesion volume was significantly decreased by 30% and 50%, respectively. Multidose APC, but not single-dose APC, increased new blood vessel formation as shown by CD105(+)/Ki-67(+) double immunostaining by nearly 2-fold at 7 days. Multidose APC also promoted posttraumatic proliferation of neuroblasts in the subventricular zone (SVZ) and their migration from the SVZ to the perilesional area., Conclusion: Activated protein C improves functional outcome and is neuroprotective after TBI. It also promotes angiogenesis and survival and migration of neuroblasts from the SVZ to the perilesional area, but the exact role of these brain repair mechanisms remains to be determined. The present findings suggest that APC therapy may hold a significant therapeutic potential for TBI.

Activated protein C (APC) is a signaling protease with anticoagulant activity. Here, we have used mice expressing a mutation in superoxide dismutase-1 (SOD1) that is linked to amyotrophic lateral sclerosis (ALS) to show that administration of APC or APC analogs with reduced anticoagulant activity after disease onset slows disease progression and extends survival. A proteolytically inactive form of APC with reduced anticoagulant activity provided no benefit. APC crossed the blood-spinal cord barrier in mice via endothelial protein C receptor. When administered after disease onset, APC eliminated leakage of hemoglobin-derived products across the blood-spinal cord barrier and delayed microglial activation. In microvessels, motor neurons, and microglial cells from SOD1-mutant mice and in cultured neuronal cells, APC transcriptionally downregulated SOD1. Inhibition of SOD1 synthesis in neuronal cells by APC required protease-activated receptor-1 (PAR1) and PAR3, which inhibited nuclear transport of the Sp1 transcription factor. Diminished mutant SOD1 synthesis by selective gene excision within endothelial cells did not alter disease progression, which suggests that diminished mutant SOD1 synthesis in other cells, including motor neurons and microglia, caused the APC-mediated slowing of disease. The delayed disease progression in mice after APC administration suggests that this approach may be of benefit to patients with familial, and possibly sporadic, ALS.

Brain hemorrhage is a serious complication of tissue plasminogen activator (tPA) therapy for ischemic stroke. Here we report that activated protein C (APC), a plasma serine protease with systemic anticoagulant, anti-inflammatory and antiapoptotic activities, and direct vasculoprotective and neuroprotective activities, blocks tPA-mediated brain hemorrhage after transient brain ischemia and embolic stroke in rodents. We show that APC inhibits a pro-hemorrhagic tPA-induced, NF-kappaB-dependent matrix metalloproteinase-9 pathway in ischemic brain endothelium in vivo and in vitro by acting through protease-activated receptor 1. The present findings suggest that APC may improve thrombolytic therapy for stroke, in part, by reducing tPA-mediated hemorrhage.

Recent studies suggest that moderate red wine consumption reduces the incidence of Alzheimer's disease (AD) clinical dementia. Using Tg2576 mice, which model AD-type amyloid beta-protein (Abeta) neuropathology, we tested whether moderate consumption of the red wine Cabernet Sauvignon modulates AD-type neuropathology and cognitive deterioration. The wine used in the study was generated using Cabernet Sauvignon grapes from Fresno, California, and was delivered to Tg2576 in a final concentration of approximately 6% ethanol. We found that Cabernet Sauvignon significantly attenuated AD-type deterioration of spatial memory function and Abeta neuropathology in Tg2576 mice relative to control Tg2576 mice that were treated with either a comparable amount of ethanol or water alone. Chemical analysis showed the Cabernet Sauvignon used in this study contains a very low content of resveratrol (0.2 mg/L), 10-fold lower than the minimal effective concentration shown to promote Abeta clearance in vitro. Our studies suggest Cabernet Sauvignon exerts a beneficial effect by promoting nonamyloidogenic processing of amyloid precursor protein, which ultimately prevents the generation of Abeta peptides. This study supports epidemiological evidence indicating that moderate wine consumption, within the range recommended by the FDA dietary guidelines of one drink per day for women and two for men, may help reduce the relative risk for AD clinical dementia.

Nicotinamide adenine dinucleotide (NAD)+-dependent sirtuins have been identified to be key regulators in the lifespan extending effects of calorie restriction (CR) in a number of species. In this study we report for the first time that promotion of the NAD+-dependent sirtuin, SIRT1-mediated deacetylase activity, may be a mechanism by which CR influences Alzheimer disease (AD)-type amyloid neuropathology. Most importantly, we report that the predicted attenuation of beta-amyloid content in the brain during CR can be reproduced in mouse neurons in vitro by manipulating cellular SIRT1 expression/activity through mechanisms involving the regulation of the serine/threonine Rho kinase ROCK1, known in part for its role in the inhibition of the non-amyloidogenic alpha-secretase processing of the amyloid precursor protein. Conversely, we found that the expression of constitutively active ROCK1 in vitro cultures significantly prevented SIRT1-mediated response, suggesting that alpha-secretase activity is required for SIRT1-mediated prevention of AD-type amyloid neuropathology. Consistently we found that the expression of exogenous human (h) SIRT1 in the brain of hSIRT1 transgenics also resulted in decreased ROCK1 expression and elevated alpha-secretase activity in vivo. These results demonstrate for the first time a role for SIRT1 activation in the brain as a novel mechanism through which CR may influence AD amyloid neuropathology. The study provides a potentially novel pharmacological strategy for AD prevention and/or treatment.

Background: The cause of neuronal death in amyotrophic lateral sclerosis (ALS) is uncertain but mitochondrial dysfunction may play an important role. Ketones promote mitochondrial energy production and membrane stabilization., Results: SOD1-G93A transgenic ALS mice were fed a ketogenic diet (KD) based on known formulations for humans. Motor performance, longevity, and motor neuron counts were measured in treated and disease controls. Because mitochondrial dysfunction plays a central role in neuronal cell death in ALS, we also studied the effect that the principal ketone body, D-beta-3 hydroxybutyrate (DBH), has on mitochondrial ATP generation and neuroprotection. Blood ketones were > 3.5 times higher in KD fed animals compared to controls. KD fed mice lost 50% of baseline motor performance 25 days later than disease controls. KD animals weighed 4.6 g more than disease control animals at study endpoint; the interaction between diet and change in weight was significant (p = 0.047). In spinal cord sections obtained at the study endpoint, there were more motor neurons in KD fed animals (p = 0.030). DBH prevented rotenone mediated inhibition of mitochondrial complex I but not malonate inhibition of complex II. Rotenone neurotoxicity in SMI-32 immunopositive motor neurons was also inhibited by DBH., Conclusion: This is the first study showing that diet, specifically a KD, alters the progression of the clinical and biological manifestations of the G93A SOD1 transgenic mouse model of ALS. These effects may be due to the ability of ketone bodies to promote ATP synthesis and bypass inhibition of complex I in the mitochondrial respiratory chain.

To fulfill the SIA roadmap requirements for EUV resists, the development of entirely new polymer platforms is necessary. In order to address issues like Line Edge Roughness (LER) and photospeed, we have developed a novel chemically amplified photoresist containing a photoacid generator (PAG) in the main chain of the polymer. The incorporation of a cationic PAG unit, phenyl methacrylate dimethylsulfonium nonaflate (PAG), in the resist backbone showed increased sensitivity, when compared with analogous blend PAG resist samples. In addition, the overall lithographic performance improved by using the counter anion (nonaflate) in the PAG units. The newly synthesized polymer bound PAG resist, poly (4-hydroxystyrene-co-2-ethyl-2-adamantyl methacrylate-co-PAG) showed sub-50 nm features using EUV Lithography.

Object: The authors evaluated the neuroprotective effect of 5,10,15,20-tetrakis(N-methyl-4'-pyridyl)porphyrinato-iron(III) (FeTMPyP), a peroxynitrite decomposition catalyst, and 1,5-isoquinolinediol (ISO), a poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) inhibitor, alone and in combination in rats with focal cerebral ischemia induced by middle cerebral artery occlusion (MCAO)., Methods: Male Sprague-Dawley rats were subjected to 2 hours of MCAO followed by 22 hours of reperfusion. Cerebral infarction and neurological deficits were estimated after ischemia. Intraperitoneal injections of FeTMPyP (1 and 2 mg/kg) and ISO (0.05 and 0.1 mg/kg) were administered alone or in combination in ischemic animals. The PARP activity in vehicle- and drug-treated groups was estimated using anti-poly(ADP-ribose) antibody in immunofluorescence and immunoblotting studies. Two hours of MCAO and 22 hours of reperfusion produced significant cerebral infarction and neurological deficits. Treatment with FeTMPyP (1 and 2 mg/kg) and ISO (0.05 and 0.1 mg/kg) produced a significant reduction in cerebral infarction and neurological deficits. Combination therapy (2 mg/kg FeTMPyP and 0.1 mg/kg ISO) enhanced the inhibition of ischemic volume (77.81+/-0.86%) compared with monotherapies (FeTMPyP 54.07+/-5.6% and ISO 53.06+/-3.88%). Immunoblotting and immunofluorescence studies showed PARP activation after ischemia, which was reduced by drug treatment., Conclusions: Neuroprotection observed with FeTMPyP and ISO alone and in combination may be attributed to inhibition of the peroxynitrite-PARP cascade of cerebral ischemia/reperfusion injury.

Free radicals have been implicated in cerebral ischemia reperfusion (IR) injury. Massive production of nitric oxide and superoxide results in continuous formation of peroxynitrite even several hours after IR insult. This can produce DNA strand nicks, hydroxylation and/or nitration of cytosolic components of neuron, leading to neuronal death. Peroxynitrite decomposition catalysts 5,10,15,20-tetrakis(N-methyl-4'-pyridyl)porphyrinato iron (III) (FeTMPyP) and 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrinato iron (III) (FeTPPS) have been demonstrated to protect neurons in in vitro cultures; however, their neuroprotective efficacy in cerebral IR injury has not been explored. In the present study, we investigated the efficacy and the therapeutic time window of FeTMPyP and FeTPPS in focal cerebral ischemia (FCI). FCI was induced according to the middle cerebral artery occlusion (MCAO) method. After 2 h of MCAO and 70 h of reperfusion, the extent of neurological deficits, infarct and edema volume were measured in Sprague-Dawley rats. FeTMPyP and FeTPPS were administered at different time points 2, 6, 9 and 12 h post MCAO. FeTMPyP and FeTPPS (3 mg kg(-1), i.v.) treatment at 2 and 6 h post MCAO produced significant reduction in infarct volume, edema volume and neurological deficits. However, treatment at latter time points did not produce significant neuroprotection. Significant reduction of peroxynitrite in blood and nitrotyrosine in brain sections was observed on FeTMPyP and FeTPPS treatment. As delayed treatment of FeTMPyP and FeTPPS produced neuroprotection, we tested whether treatment had any influence over the apoptotic neuronal death. DNA fragmentation and in situ nick end-labeling assays showed that FeTMPyP and FeTPPS treatment reduced IR injury-induced DNA fragmentation. In conclusion, peroxynitrite decomposition catalysts (FeTMPyP and FeTPPS) produced prominent neuroprotection even if administered 6 h post MCAO and the neuroprotective effect is at least in part due to the reduction of peroxynitrite and apoptosis.

In the present study, neuroprotective effect of 4-amino-1,8-napthalimide (4-ANI), a poly(ADP-ribose) polymerase (PARP) inhibitor was investigated in middle cerebral artery occlusion (MCAo)-induced focal ischemia. Sprague-Dawley rats were subjected to 2 h of middle cerebral artery occlusion followed by 22 h of reperfusion. After 22 h of reperfusion rats were evaluated for cerebral infarction, neurological deficits, brain NAD levels, and in situ terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL). Focal ischemia produced significant infarct volume (201 +/- 14 mm3), neurological scores (2 +/- 0.5) and 28 +/- 4.5% brain NAD depletion. Ischemia was associated with increased in TUNEL positive cells in brain sections indicating DNA fragmentation. 4-ANI treatment (1 and 3 mg/kg, i.p.) significantly decreased infarct volume to 35 +/- 7% and 70 +/- 6%, respectively. Neurological functions were also significantly improved at these doses. 4-ANI (3 mg/kg) completely reversed brain NAD depletion and significantly reduced the increase in the number of TUNEL positive cells. Nevertheless, 4-ANI treatment did not alter cerebral blood flow and blood pressure. Our study suggests 4-ANI is a potent neuroprotective agent in focal cerebral ischemia and its neuroprotective effects may be attributed to reduction of NAD depletion and DNA fragmentation.

Free radical induced neuronal damage is implicated in cerebral ischemia reperfusion (IR) injury and antioxidants are reported to have neuroprotective activity. Several in vitro and in vivo studies have proved the antioxidant potential of curcumin and its metabolites. Hence, in the present study the neuroprotective potential of curcumin was investigated in middle cerebral artery occlusion (MCAO) induced focal cerebral IR injury. 2 h of MCAO and 22 h of reperfusion resulted in the infarct volume of 210.39 +/- 31.25 mm3. Administration of curcumin 100 and 300 mg/kg, i.p. 30 min. after MCAO produced 37.23 +/- 5.10% and 46.39 +/- 10.23% (p < 0.05) reduction in infarct volume, respectively. Ischemia induced cerebral edema was reduced in a dose dependent manner. Curcumin at 300 mg/kg, i.p. produced 50.96 +/- 6.04% reduction in edema (p < 0.05) volume. Increase in lipid peroxidation after MCAO in ipsilateral and contralateral hemisphere of brain was observed, which was reduced by curcumin (300 mg/kg, i.p.)-treatment. Decrease in superoxide dismutase and glutathione peroxidase activity was observed in ipsilateral hemisphere of MCAO animal. Curcumin-treatment (300 mg/kg, i.p.) prevented IR injury mediated fall in glutathione peroxide activity. Peroxynitrite measured using rhodamine123 fluorescence and anti-nitrotyrosine immunofluorescence indicated increased peroxynitrite formation after IR insult. Curcumin-treatment reduced peroxynitrite formation and hence the extent of tyrosine nitration in the cytosolic proteins. These results suggest the neuroprotective potential of curcumin in cerebral ischemia and is mediated through its antioxidant activity.

A novel template guided enzymatic approach has been developed to synthesize optically active conducting polyaniline (PANI) nanocomposites in the presence of H2O2 as an oxidant, using (+) and (-) 10-camphorsulfonic acid (CSA) as a dopant and chiral inductor. The formation of chiral polyaniline in the nanocomposites was confirmed by circular dichroism (CD). Interestingly, the CD spectra of nanocomposites formed either with (+) or with (-) CSA show the enzyme itself plays a critical role in controlling the stereospecificity of the polyaniline (PANI) in the nanocomposite. The enzyme used for the polymerization of aniline in the nanocomposite was horseradish peroxidase (HRP). It was shown that this enzyme prefers a specific helical conformation, regardless of whether induced chirality in the complex CSA-aniline is from (+) or (-) CSA. UV-vis spectra show that the polyaniline is in the conducting form, and transmission electron micrographs (TEM) show that the nanocomposites are dispersed nicely with particle size dimensions in the range of 20-50 nm. Electron diffraction patterns of these chiral polymer nanocomposites suggest that these nanocomposites are in both crystalline and amorphous states.

The present study was undertaken to investigate the effect of testosterone on the alpha-adrenoceptor-mediated contractile responses in ventral lobe of rat prostate. Contractile responses to various alpha-adrenoceptor agonists (phenylephrine, A61603 (N-[5-(4,5-dihydro-1H-imidazol-2-yl)-2-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl] methanesulphonamide), clonidine, guanfacine, ST587 ((2,3-dihydro-benzo[1,4]dioxin-2-ylmethyl)-[2-(2,6-dimethoxy-phenoxy)-ethyl]-amine) and xylazine) were tested in prostate strips obtained from control and testosterone (3 mg/kg, s.c. 5 days a week for 15 days-10 doses total)-treated rats. Dose-response curves for alpha-adrenoceptor agonists in testosterone-treated animals showed a leftward shift, indicating increased sensitivity of tissue to alpha-adrenoceptor agonists. To find the mechanism of increased sensitivity, K(A) value and receptor reserve of phenylephrine were estimated. Neither the K(A) value nor the receptor reserve of phenylephrine was altered in testosterone-treated rats. The concentration-occupancy curve for A61603 was shifted leftward and the K(A) value for A61603 decreased about four-fold. The K(B) value of 2-(2,6-dimethoxyphenoxyethyl) aminomethyl-1,4-benzodioxane (WB4101) was not altered, however, the K(B) value for prazosin was decreased approximately 5.5-fold. These findings indicate that the testosterone-mediated increase in sensitivity of prostate to alpha-adrenoceptor agonists is due to alterations in the alpha(1)-adrenoceptor pool.

Benign prostate hyperplasia (BPH) is a common malady affecting elderly men throughout the world, and it is the most common cause of voiding dysfunctions in man. The disease becomes prevalent, as the mean age of the population increases. In BPH the glandular cells and myofibroblasts of periurethral and transition zones proliferate excessively. This excessive growth of tissue mass physically compresses the urethra, leading to urinary obstruction and lower urinary tract symptoms (LUTS). Another major factor that contributes to LUTS in BPH is the increase in smooth muscle tone, a dynamic component. Contraction of the prostate gland is mainly under the control of the autonomic system and is mediated through alpha-adrenoceptors. In addition to alpha1-adrenoceptors, there are many other components that influence growth and contraction of the prostate gland, resulting in the development of BPH and LUTS. The prostate gland is contracted on stimulating 5-HT, endothelin, tachykinin, and muscarinic cholinergic receptors. Growth of the prostate gland is under the control of androgens, endothelin growth factor, vasoactive intestinal peptide, nerve growth factor, and growth hormone. The combination of excessive growth and the tone produced by different components results in LUTS and BPH. alpha1-Adrenoceptor antagonists were successfully used in the treatment of BPH to relieve the LUTS. The high selectivity of alpha1A-adrenoceptor antagonists reflects the uroselectivity. The uroselective compounds like terazosin, doxazosin, tamsulosin, and alfuzosin are in clinical use. Other new potent uroselective compounds, which are analogs of nigulidipine, 5-methyl-urapidil, or naftopidil, are in clinical trials. In addition to alpha-antagonists, antiandrogens (5alpha-reductase inhibitors) and polyherbal formulations are used to treat BPH. The success of BPH treatment lies in the development of new chemical entities which are efficacious as well as more uroselective and hence have least side effects., (Copyright 2002 S. Karger AG, Basel)

Mutation of Galpha(q) or Galpha(s) N-terminal contact sites for Gbetagamma resulted in alpha subunits that failed to localize at the plasma membrane or undergo palmitoylation when expressed in HEK293 cells. We now show that overexpression of specific betagamma subunits can recover plasma membrane localization and palmitoylation of the betagamma-binding-deficient mutants of alpha(s) or alpha(q). Thus, the betagamma-binding-defective alpha is completely dependent on co-expression of exogenous betagamma for proper membrane localization. In this report, we examined the ability of beta(1-5) in combination with gamma(2) or gamma(3) to promote proper localization and palmitoylation of mutant alpha(s) or alpha(q). Immunofluorescence localization, cellular fractionation, and palmitate labeling revealed distinct subtype-specific differences in betagamma interactions with alpha subunits. These studies demonstrate that 1) alpha and betagamma reciprocally promote the plasma membrane targeting of the other subunit; 2) beta(5), when co-expressed with gamma(2) or gamma(3), fails to localize to the plasma membrane or promote plasma membrane localization of mutant alpha(s) or alpha(q); 3) beta(3) is deficient in promoting plasma membrane localization of mutant alpha(s) and alpha(q), whereas beta(4) is deficient in promoting plasma membrane localization of mutant alpha(q); 4) both palmitoylation and interactions with betagamma are required for plasma membrane localization of alpha.

The binding affinity of purified native Ustilago maydis topoisomerase I enzyme for radiolabeled DNA substrates with various secondary structures was determined by gel shift and equilibrium binding analysis. Topoisomerase I exhibited cooperativity in binding to DNA regardless of the substrate structure. Further analysis demonstrated that cruciform DNA has two populations of binding sites for topoisomerase I while the other substrates (single-stranded DNA, DNA molecules containing six or one mismatched base pairs, hairpin, and fully homologous duplex DNA) have a single population of binding sites. The affinity of topoisomerase I for cruciform was found to be an order of magnitude higher affinity than for any of the other substrates. The high affinity of topoisomerase I for cruciform and specificity of topoisomerase I-cruciform structure interaction were confirmed by competition experiments. These studies demonstrate the high affinity of topoisomerase I for cruciform structure.

The Ustilago maydis genomic TOP1 gene encoding DNA topoisomerase I was cloned by amplifying a gene fragment using the polymerase chain reaction, and using this fragment to search a genomic DNA library by hybridization. The predicted peptide sequence exhibited 30-40% identity to other eukaryotic TOP1 genes, yet differed in several features. First, an unusually long acidic region was identified near the amino terminus (28/29 residues are acidic), which resembles other nucleolar peptide motifs. Second, an atypical carboxy-terminal 'tail', absent in other TOP1 genes, followed the active site tyrosine residue. A top1 gene disruption mutant was constructed by replacing the genomic TOP1 gene, with a top1::HygR null allele. This mutant lost the abundant topoisomerase I activity evident in wild-type U.maydis, and displayed a subtle coloration phenotype evident during cell senescence.

Ustilago maydis topoisomerase I relaxes superhelical DNA in the absence of any co-factors. The reaction reaches a defined end-point proportional to the amount of enzyme added and an analysis of the reaction by Hill plot transformation indicates that at least two molecules of topoisomerase must interact with the DNA to catalyze relaxation. The addition of purified Ustilago histone H1 reduces the stoichiometric amount of topoisomerase I required by 50%. H1 histone may function to enhance DNA relaxation through a cooperative mechanism. The purified HMG-like protein from Ustilago also enhances DNA relaxation mediated by the topoisomerase. Whereas H1 stimulates topo I-mediated DNA relaxation through a processive mode, the HMG-like protein enhances through a distributive mechanism. Taken together, these results demonstrate that the interaction of chromosomal proteins with topoisomerase can influence DNA topology, and mechanisms are proposed to explain this enhancement.