Delayed Cerebral Infarction Following Trans-Sylvian Surgery for Craniopharyngioma Presenting as Status Epilepticus.

The postoperative period after craniopharyngioma surgery has a high likelihood of complications and its management can be challenging. We present the case of a 10-year-old boy who was operated for craniopharyngioma. In the postoperative period he developed lacunar infarct, endocrine disturbances, delayed vasospasm leading to cerebral infarction and status epilepticus. The likely cause of status epilepticus was likely delayed cerebral infarction compounded with dyselectrolytemia. We discuss the possible etiology of delayed cerebral infarction.

Aryl Hydrocarbon Receptor Mediates Larval Zebrafish Fin Duplication Following Exposure to Benzofluoranthenes.

The aryl hydrocarbon receptor (AHR) mediates developmental toxicity of several xenobiotic classes including polycyclic aromatic hydrocarbons. Using embryonic zebrafish, we previously identified 4 polycyclic aromatic hydrocarbons that caused a novel phenotype among AHR ligands-growth of a lateral, duplicate caudal fin fold. The window of sensitivity to the most potent inducer of this phenotype, benzo[k]fluoranthene (BkF), was prior to 36 h postfertilization (hpf), although the phenotype was not manifest until 60 hpf. AHR dependency via Ahr2 was demonstrated using morpholino knockdown. Hepatocyte ablation demonstrated that hepatic metabolism of BkF was not required for the phenotype, nor was it responsible for the window of sensitivity. RNA sequencing performed on caudal trunk tissue from BkF-exposed animals collected at 48, 60, 72, and 96 hpf showed upregulation of genes associated with AHR activation, appendage development, and tissue patterning. Genes encoding fibroblast growth factor and bone morphogenic protein ligands, along with retinaldehyde dehydrogenase, were prominently upregulated. Gene Ontology term analysis revealed that upregulated genes were enriched for mesoderm development and fin regeneration, whereas downregulated genes were enriched for Wnt signaling and neuronal development. MetaCore (Clarivate Analytics) systems analysis of orthologous human genes predicted that R-SMADs, AP-1, and LEF1 regulated the expression of an enriched number of gene targets across all time points. Our results demonstrate a novel aspect of AHR activity with implications for developmental processes conserved across vertebrate species.

Coupling Genome-wide Transcriptomics and Developmental Toxicity Profiles in Zebrafish to Characterize Polycyclic Aromatic Hydrocarbon (PAH) Hazard.

Polycyclic Aromatic Hydrocarbons (PAHs) are diverse environmental pollutants associated with adverse human health effects. Many studies focus on the carcinogenic effects of a limited number of PAHs and there is an increasing need to understand mechanisms of developmental toxicity of more varied yet environmentally relevant PAHs. A previous study characterized the developmental toxicity of 123 PAHs in zebrafish. Based on phenotypic responses ranging from complete inactivity to acute mortality, we classified these PAHs into eight bins, selected 16 representative PAHs, and exposed developing zebrafish to the concentration of each PAH that induced 80% phenotypic effect. We conducted RNA sequencing at 48 h post fertilization to identify gene expression changes as a result of PAH exposure. Using the Context Likelihood of Relatedness algorithm, we inferred a network that links the PAHs based on coordinated gene responses to PAH exposure. The 16 PAHs formed two broad clusters: Cluster A was transcriptionally more similar to the controls, while Cluster B consisted of PAHs that were generally more developmentally toxic, significantly elevated cyp1a transcript levels, and induced Ahr2-dependent Cyp1a protein expression in the skin confirmed by gene-silencing studies. We found that cyp1a transcript levels were associated with transcriptomic response, but not with PAH developmental toxicity. While all cluster B PAHs predominantly activated Ahr2, they also each enriched unique pathways like ion transport signaling, which likely points to differing molecular events between the PAHs downstream of Ahr2. Thus, using a systems biology approach, we have begun to evaluate, classify, and define mechanisms of PAH toxicity.

Systematic developmental neurotoxicity assessment of a representative PAH Superfund mixture using zebrafish.

Superfund sites often consist of complex mixtures of polycyclic aromatic hydrocarbons (PAHs). It is widely recognized that PAHs pose risks to human and environmental health, but the risks posed by exposure to PAH mixtures are unclear. We constructed an environmentally relevant PAH mixture with the top 10 most prevalent PAHs (SM10) from a Superfund site derived from environmental passive sampling data. Using the zebrafish model, we measured body burden at 48 hours post fertilization (hpf) and evaluated the developmental and neurotoxicity of SM10 and the 10 individual constituents at 24 hours post fertilization (hpf) and 5 days post fertilization (dpf). Zebrafish embryos were exposed from 6 to 120 hpf to (1) the SM10 mixture, (2) a variety of individual PAHs: pyrene, fluoranthene, retene, benzo[a]anthracene, chrysene, naphthalene, acenaphthene, phenanthrene, fluorene, and 2-methylnaphthalene. We demonstrated that SM10 and only 3 of the individual PAHs were developmentally toxic. Subsequently, we constructed and exposed developing zebrafish to two sub-mixtures: SM3 (comprised of 3 of the developmentally toxicity PAHs) and SM7 (7 non-developmentally toxic PAHs). We found that the SM3 toxicity profile was similar to SM10, and SM7 unexpectedly elicited developmental toxicity unlike that seen with its individual components. The results demonstrated that the overall developmental toxicity in the mixtures could be explained using the general concentration addition model. To determine if exposures activated the AHR pathway, spatial expression of CYP1A was evaluated in the 10 individual PAHs and the 3 mixtures at 5 dpf. Results showed activation of AHR in the liver and vasculature for the mixtures and some individual PAHs. Embryos exposed to SM10 during development and raised in chemical-free water into adulthood exhibited decreased learning and responses to startle stimulus indicating that developmental SM10 exposures affect neurobehavior. Collectively, these results exemplify the utility of zebrafish to investigate the developmental and neurotoxicity of complex mixtures.

Comparative developmental toxicity of a comprehensive suite of polycyclic aromatic hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants that occur in complex mixtures. Several PAHs are known or suspected mutagens and/or carcinogens, but developmental toxicity data is lacking for PAHs, particularly their oxygenated and nitrated derivatives. Such data are necessary to understand and predict the toxicity of environmental mixtures. 123 PAHs were assessed for morphological and neurobehavioral effects for a range of concentrations between 0.1 and 50 µM, using a high throughput early-life stage zebrafish assay, including 33 parent, 22 nitrated, 17 oxygenated, 19 hydroxylated, 14 methylated, 16 heterocyclic, and 2 aminated PAHs. Additionally, each PAH was evaluated for AHR activation, by assessing CYP1A protein expression using whole animal immunohistochemistry (IHC). Responses to PAHs varied in a structurally dependent manner. High-molecular weight PAHs were significantly more developmentally toxic than the low-molecular weight PAHs, and CYP1A expression was detected in five distinct tissues, including vasculature, liver, skin, neuromasts and yolk.

Aerobic Bioremediation of PAH Contaminated Soil Results in Increased Genotoxicity and Developmental Toxicity.

The formation of more polar and toxic polycyclic aromatic hydrocarbon (PAH) transformation products is one of the concerns associated with the bioremediation of PAH-contaminated soils. Soil contaminated with coal tar (prebioremediation) from a former manufactured gas plant (MGP) site was treated in a laboratory scale bioreactor (postbioremediation) and extracted using pressurized liquid extraction. The soil extracts were fractionated, based on polarity, and analyzed for 88 PAHs (unsubstituted, oxygenated, nitrated, and heterocyclic PAHs). The PAH concentrations in the soil tested, postbioremediation, were lower than their regulatory maximum allowable concentrations (MACs), with the exception of the higher molecular weight PAHs (BaA, BkF, BbF, BaP, and IcdP), most of which did not undergo significant biodegradation. The soil extract fractions were tested for genotoxicity using the DT40 chicken lymphocyte bioassay and developmental toxicity using the embryonic zebrafish (Danio rerio) bioassay. A statistically significant increase in genotoxicity was measured in the unfractionated soil extract, as well as in four polar soil extract fractions, postbioremediation (p < 0.05). In addition, a statistically significant increase in developmental toxicity was measured in one polar soil extract fraction, postbioremediation (p < 0.05). A series of morphological abnormalities, including peculiar caudal fin malformations and hyperpigmentation in the tail, were measured in several soil extract fractions in embryonic zebrafish, both pre- and postbioremediation. The increased toxicity measured postbioremediation is not likely due to the 88 PAHs measured in this study (including quinones), because most were not present in the toxic polar fractions and/or because their concentrations did not increase postbioremediation. However, the increased toxicity measured postbioremediation is likely due to hydroxylated and carboxylated transformation products of the 3- and 4-ring PAHs (PHE, 1MPHE, 2MPHE, PRY, BaA, and FLA) that were most degraded.

Non-instrumented incubation of a recombinase polymerase amplification assay for the rapid and sensitive detection of proviral HIV-1 DNA.

Sensitive diagnostic tests for infectious diseases often employ nucleic acid amplification technologies (NAATs). However, most NAAT assays, including many isothermal amplification methods, require power-dependent instrumentation for incubation. For use in low resource settings (LRS), diagnostics that do not require consistent electricity supply would be ideal. Recombinase polymerase amplification (RPA) is an isothermal amplification technology that has been shown to typically work at temperatures ranging from 25-43°C, and does not require a stringent incubation temperature for optimal performance. Here we evaluate the ability to incubate an HIV-1 RPA assay, intended for use as an infant HIV diagnostic in LRS, at ambient temperatures or with a simple non-instrumented heat source. To determine the range of expected ambient temperatures in settings where an HIV-1 infant diagnostic would be of most use, a dataset of the seasonal range of daily temperatures in sub Saharan Africa was analyzed and revealed ambient temperatures as low as 10°C and rarely above 43°C. All 24 of 24 (100%) HIV-1 RPA reactions amplified when incubated for 20 minutes between 31°C and 43°C. The amplification from the HIV-1 RPA assay under investigation at temperatures was less consistent below 30°C. Thus, we developed a chemical heater to incubate HIV-1 RPA assays when ambient temperatures are between 10°C and 30°C. All 12/12 (100%) reactions amplified with chemical heat incubation from ambient temperatures of 15°C, 20°C, 25°C and 30°C. We also observed that incubation at 30 minutes improved assay performance at lower temperatures where detection was sporadic using 20 minutes incubation. We have demonstrated that incubation of the RPA HIV-1 assay via ambient temperatures or using chemical heaters yields similar results to using electrically powered devices. We propose that this RPA HIV-1 assay may not need dedicated equipment to be a highly sensitive tool to diagnose infant HIV-1 in LRS.

Multidimensional in vivo hazard assessment using zebrafish.

There are tens of thousands of man-made chemicals in the environment; the inherent safety of most of these chemicals is not known. Relevant biological platforms and new computational tools are needed to prioritize testing of chemicals with limited human health hazard information. We describe an experimental design for high-throughput characterization of multidimensional in vivo effects with the power to evaluate trends relating to commonly cited chemical predictors. We evaluated all 1060 unique U.S. EPA ToxCast phase 1 and 2 compounds using the embryonic zebrafish and found that 487 induced significant adverse biological responses. The utilization of 18 simultaneously measured endpoints means that the entire system serves as a robust biological sensor for chemical hazard. The experimental design enabled us to describe global patterns of variation across tested compounds, evaluate the concordance of the available in vitro and in vivo phase 1 data with this study, highlight specific mechanisms/value-added/novel biology related to notochord development, and demonstrate that the developmental zebrafish detects adverse responses that would be missed by less comprehensive testing strategies.

The extrinsic origin of the magnetodielectric effect in the double perovskite La2NiMnO6.

A La(2)NiMnO(6) polycrystalline sample prepared by the sol-gel method showed monoclinic crystal structure with the P2(1)/n space group and a saturation magnetization of 4.63 µ(B)/f.u. at 5 K. Impedance spectroscopy results in the temperature range of 10 K < T < 300 K have revealed a distinct conduction process at grains and grain boundaries, where the grains followed the variable range hopping mechanism and the grain boundaries obeyed Arrhenius thermal activation. A negative magnetoresistance of 2.5% was observed at the paramagnetic to ferromagnetic transition, and this became temperature independent below the magnetic ordering. A marginal positive magnetodielectric (MD) effect that followed the dielectric relaxation was observed and its magnitude was found to decrease with increase of the frequency. A systematic study on the magnetic field induced dielectric properties, dc transport and dc bias effect on the dielectric permittivity has revealed the extrinsic origin of the MD effect in the bulk sample of La(2)NiMnO(6).

Site-selective synthesis of in situ Ni-filled multi-walled carbon nanotubes using Ni(salen) as a catalyst source.

The synthesis of Ni-filled multi-walled carbon nanotubes was performed by atmospheric pressure chemical vapor deposition with propane on Si at 850 °C using a simple mixture of (N, N'-bis(salicylidene)-ethylenediiminato) nickel(II), commonly known as Ni(salen), and a conventional photoresist. Analysis of the carbon nanotubes using scanning electron microscopy together with high-resolution transmission electron microscopy show that the nanotubes have grown by a tip-growth mechanism and exhibit a multi-walled structure with partial Ni filling. The high quality of the Ni-filled nanotubes is evidenced by Raman spectroscopy. The magnetic properties of Ni-filled nanotubes were analyzed using a superconducting quantum interference device which revealed their ferromagnetic behavior with large coercivity. A scalable as well as site-selective growth of high quality Ni-filled carbon nanotubes is achieved by a simple photolithographic method.

Role of peritoneal macrophages and lymphocytes in the development of hypogonadal osteoporosis in an ovariectomized rat model: possible phytoestrogenic efficacy of oil extract of garlic to preserve skeletal health.

This study was to examine whether skeletal health deterioration in the hypogonadal situation is a consequence of an alteration in the functional status of peripheral mononuclear cells and its amelioration, if any, by an oil extract of garlic. The results suggest that hypogonadism-induced oxidative stress of peritoneal macrophages and lymphocytes could be reduced by supplementation with an oil extract of garlic. However, estrogen deficiency did not cause any significant change in DNA fragmentation of peritoneal macrophages. The hypogonadism-induced increase in the serum levels of IL-6 and TNF-alpha were significantly reduced by an oil extract of garlic. Further, such supplementation could revive the hypogonadism-induced decrease in serum estrogen titer and counter-balance the increase in bone turnover as determined by low bone tensile strength and alterations in bone related biochemical variables such as urinary calcium, hydroxyproline, calcium to creatinine ratio and serum tartrate resistant acid phosphatase activity (TRAP). The garlic oil supplemented partial recovery of the serum estrogen titer in hypogonadal rats was found to be persistently associated with reduced oxidative stress of peritoneal macrophages and lymphocytes, reduced serum interleukins and better preservation of bone mass. This study proposes that the hypogonadism-induced bone loss has a direct correlation with the functional status of lymphocytes and peritoneal macrophages, and garlic can prevent this.

Effects of garlic oil on postmenopausal osteoporosis using ovariectomized rats: comparison with the effects of lovastatin and 17beta-estradiol.

The purpose of this study was to examine the antiosteoporosis effects of garlic oil in an ovariectomized (Ovx) rat model of osteoporosis and to compare its efficacy with lovastatin (a synthetic hypocholesterolemic drug) and 17beta-estradiol (a potent antiosteoporotic agent). Animals were divided into five groups: sham-operated control, ovariectomized, ovariectomized supplemented with lovastatin, ovariectomized supplemented with garlic oil and ovariectomized supplemented with 17beta-estradiol. In our study, the development of a high rate of bone turnover and osteoporosis in the ovariectomized animals were confirmed by significant alterations of serum alkaline phosphatase activity, serum tartrate-resistant acid phosphatase activity, urinary excretion of calcium, phosphate, hydroxyproline and urinary calcium to creatinine ratio, when compared with the sham-operated control group. Supplementation of these animals with either garlic oil or lovastatin or 17beta-estradiol, in addition to their hypocholesterolemic effect, could counterbalance all these changes. The results revealed that all three compounds significantly protected the hypogonadal bone loss as reflected by higher bone densities and higher bone mineral contents than the ovariectomized group of animals. The results emphasize that, like 17beta-estradiol, the hypocholesterolemic compounds garlic oil and lovastatin are also effective in suppressing bone loss owing to estrogen deficiency and their efficacy in the order of lower to higher is garlic < lovastatin < 17beta-estradiol.

Fractal studies on the protein secondary structure elements.

The fractal dimensions of the protein secondary structure elements based on the positional distributions were calculated. The seven secondary structure elements could be grouped into two distinct classes based on their fractal dimensions. A negative correlation was observed between the dimensional values of the secondary structures and their abundances. Markov model was applied, to check the persistence of transitions of the secondary structure elements, in the protein sequences. The results suggest the presence of long-range correlations in the protein sequences that may be useful in prediction algorithms.

Prevention of bone loss by oil extract of garlic (Allium sativum Linn.) in an ovariectomized rat model of osteoporosis.

The effects of oil extract of garlic (Allium sativum Linn.) on different primary and secondary osteoporotic marker changes were tested in an ovariectomized rat model of osteoporosis. Experiments were performed on three different rat models: sham-operated control, ovariectomized and ovariectomized supplemented with garlic oil. In ovariectomized group, there has been a significant increase in different relative organ weights compared to sham-operated control, while the uterine weight was found to be decreased. Supplementation with oil extract of garlic could effectively reverse these changes. Also low bone densities that developed in the ovariectomized group were significantly recovered in the garlic oil supplemented group. In our study, the development of high rate of bone turnover and osteoporosis in the ovariectomized animals were confirmed by significant alteration of serum alkaline phosphatase activity, serum tartrate resistant acid phosphatase activity, urinary excretion of calcium, phosphate, hydroxyproline and urinary calcium to creatinine ratio, when compared with the sham-operated control group. Garlic oil extract supplementation, apart from its unique influence in lowering blood cholesterol, could also prevent ovariectomy-induced rise in all the above-mentioned marker changes. The results of this study emphasize that oil extract of garlic possibly has a positive role in suppressing ovariectomy-induced bone resorption.

Observation of minority spin character of the new electron doped manganite La0.7Ce0.3MnO3 from tunneling magnetoresistance.

We report the magnetotransport characteristics of a trilayer ferromagnetic tunnel junction built of an electron doped manganite (La0.7Ce0.3MnO3) and a hole doped manganite (La0.7Ca0.3MnO3). At low temperatures the junction exhibits a large positive tunneling magnetoresistance (TMR), irrespective of the bias voltage. At intermediate temperatures below T(C) the sign of the TMR is dependent on the bias voltage across the junction. The magnetoresistive characteristics of the junction strongly suggest that La0.7Ce0.3MnO3 is a minority spin carrier ferromagnet with a high degree of spin polarization, i.e., a transport half-metal.

Effects of different intensities of cold stress on certain physiological phenomena related to skeletal health in a hypogonadal rat model.

Intestinal transference pattern of calcium and associated changes in the activities of intestinal mucosal enzymes, rate of bone turnover and bone metabolism were evaluated in ovariectomized rats exposed to cold stress of various intensities i.e., mild (15 degrees C) or stronger (8 degrees C and 4 degrees C) for 5 min everyday for 7 consecutive days. Except mild cold stress-induced group (15 degrees C), rats of other two groups (8 degrees C and 4 degrees C) showed considerable decrease in the rate of in situ intestinal transference of calcium. Likewise, in these groups, the activities of intestinal mucosal enzymes, alkaline phosphatase (AP) and calcium ATPase (Ca2+-ATPase) were decreased significantly in all the segments of small intestine in a descending gradient. Also significant changes in bone turnover and bone resorption were confirmed in these animals by marked alterations in plasma AP activity, urinary calcium and phosphate excretion and urinary calcium to creatinine ratio (Ca: Cr). The skeletal changes were further ascertained by examining other physical and biochemical parameters of bone metabolism viz., body mass, bone density, ratio of mineral to matrix and mineral content of bone ash (calcium and phosphate) in the ovariectomized rat model. All these results suggest that stronger cold stress (8 degrees C and 4 degrees C) may be an important ecological factor in the development of earlier bone loss in hypogonadal rats.

Modeling the surface phenomena in carbon paste electrodes by low frequency impedance and double-layer capacitance measurements.

Impedance and capacitance studies have been performed with covalently coupled Glucose oxidase (GOD) enzyme, covalently coupled flavin adenine dinucleotide (FAD), reconstituted GOD enzyme and blank carbon paste electrodes to study the changes in the electrochemical interfacial properties. Impedance studies were performed using a low frequency impedance technique and the electrochemical surface capacitance was measured by a pulse technique. We have attempted to fit the experimental values to an equivalent circuit model. The Randles' cell circuit with Warburg impedance modeled well the experimental values and the behavior of the enzyme electrodes. The individual components of the model were calculated and the parameters were explained. The blank paste electrode showed a constant phase element behavior.

Effects of a high intake of unsaturated and saturated oils on intestinal transference of calcium and calcium mobilization from bone in an ovariectomized rat model of osteoporosis.

Intestinal transference of calcium and rate of bone turnover were evaluated in ovariectomized rats fed for 15 days with a high amount (30%) of lipid enriched with monounsaturated (groundnut oil), polyunsaturated (sunflower oil) and saturated (coconut oil) fatty acids. The results were compared with those for sham-operated control and ovariectomized groups fed a normal diet (7% groundnut oil). Irrespective of the saturation and unsaturation characteristics, all lipids (edible oils) used in our study considerably decreased the rate of in situ intestinal transference of calcium. Likewise, the activities of intestinal mucosal enzymes, alkaline phosphatase (AP) and calcium ATPase (Ca2+-ATPase) were decreased significantly in all the segments of the small intestine in a descending gradient. Significant changes in bone turnover and bone calcium (Ca) mobilization were confirmed in these animals by marked alterations in plasma AP activity, urinary calcium and phosphate excretion and calcium to creatinine (Ca:creatinine) ratio. Lipid supplementation (30%) in such ovariectomized rats using groundnut oil (monounsaturated), sunflower oil (polyunsaturated) or coconut oil (saturated) for 15 days further enhanced all of the above observed parameters. These results suggest that the intake of high amounts of lipids with different unsaturation and saturation characteristics may be an important factor in determining bone loss in ovariectomized rats.

Cold stress facilitates calcium mobilization from bone in an ovariectomized rat model of osteoporosis.

In an ovariectomized rat model of osteoporosis, the effects of cold stress on intestinal Ca2+ transference and rate of bone turnover were evaluated. In the ovariectomized rats, a significant reduction in intestinal transference of Ca2+ was associated with decreased activities of intestinal mucosal enzymes, alkaline phosphatase (AP), and calcium ATPase (Ca2+-ATPase) in all the different segments of small intestine in a descending gradient. The development of a high rate of bone turnover and osteoporosis in these animals was confirmed by significant alteration in plasma AP activity and calcium (Ca) level, urinary excretion of Ca and phosphate, and Ca : creatinine ratio. Cold stress in this model, apart from its unique influence in elevating plasma corticosterone and thyroid hormone level, enhanced all the above referred parameters studied in connection with intestinal transference of Ca2+, bone turnover rate, and osteoporosis. The results of this study emphasize that cold stress may have a positive influence on bone loss for an early development of hypogonadal osteoporosis in rats.

High-lipid diet intake is a possible predisposing factor in the development of hypogonadal osteoporosis.

The effects of a high-lipid diet on the intestinal transference of Ca2+ and the activities of intestinal enzymes were studied in ovariectomized rats. The plasma Ca2+ level and urinary loss of Ca2+ were also observed. Ovariectomy decreased both alkaline phosphatase (AP) and calcium ATPase (Ca(2+)-ATPase) activity, and also reduced the mucosal transference of Ca2+ in all parts of the small intestine. Although the plasma Ca2+ level did not change significantly, the urinary loss of Ca2+ was enhanced. A high-lipid diet supplement enhanced all these parameters in ovariectomized rats. It is therefore believed that a high-lipid diet may promote osteoporosis in ovariectomized rats as the result of Ca2+ transfer and enhanced urinary Ca2+ loss.