Palmitate exacerbates bisphenol A toxicity via induction of ER stress and mitochondrial dysfunction

Combined exposure to dietary nutrients and environmental chemicals may elicit significantly different physiological effects than single exposures. Exposure to dietary saturated fats and environmental toxins is a physiologically-significant dual exposure that is particularly associated with lower socioeconomic status, potentially placing these individuals at heightened risk of xenobiotic toxicities. However, no prior studies have examined interactions between specific lipids and environmental xenobiotics in modulating cellular health. Using primary mouse embryonic fibroblasts, we have discovered that prior exposure to the saturated fatty acid, palmitate, exacerbates cellular toxicity associated with the industrial plasticizer, bisphenol A (BPA). Cell death upon BPA exposure following palmitate pre-treatment was greater than that occurring with either exposure alone. Mechanistically, cell death was preceded by increased endoplasmic reticulum stress and loss of mitochondrial membrane potential in palmitate plus BPA exposed cells, leading to increased caspase-3 cleavage and subsequent apoptosis. Interestingly, inclusion of the unsaturated fatty acid, oleate, along with palmitate during the pre-treatment period completely abrogated the ER stress, mitochondrial toxicity, and cell death induced by subsequent exposure to BPA. Thus, our data identify for the first time an important interaction between a fatty acid and an environmental toxin and have implications for developing nutritional interventions to mitigate the deleterious effects of such xenobiotic exposures.     

Evaluation of physiology lectures conducted by students: comparison between evaluation by staff and students

As a peer-assisted learning process, minilectures on physiology were conducted by students. During this process, students lecture to their colleagues in the presence of faculty staff members. These lectures were evaluated by faculty staff and students simultaneously. The aim of this study was to compare feedback from faculty members and students on 66 minilectures conducted by students. Their perception of different qualities of lecture was assessed using a questionnaire. There were significant correlations between students and faculty members for many qualities of the lecture, including the speed of the lecture, retaining attention, clear introduction, and the overall quality of the lecture. However, ratings for gesture, eye contact, language usage, illustration usage, audiovisuals, voice usage, and important points stressed were significantly different between students and faculty members. Multiple regression analysis was performed to assess the degree of effect of different aspects of a lecture on its overall quality. Aspects such as gesture, eye contact, and language usage showed very low β-values, suggesting a poor contribution of these factors to the overall quality of the lecture for both students and faculty members. The speed of the lecture, retaining attention, and clear introduction were qualities that faculty members and students rated equally, and these were the main contributors to the overall quality of the lecture. Awareness about the possible discrepancy between ratings given by faculty members and students may be important when interpreting the evaluation results of formal lectures by these two groups.     

State-of-the-art therapeutic medical countermeasures for viral threat agents

In recent years, there has been an increase in the perceived threat of biological agents being used against civilian populations. This has prompted an urgent need for the development and procurement of medical countermeasures (MCMs) against highly pathogenic viruses that can prevent morbidity and mortality from infections caused by these agents. To date, antiviral drug development has been largely focused on clinically prevalent chronic infections due to their commercial viability. This has left a huge gap in the drug development path for acute infections of biodefense importance. In this review, we discuss the antiviral research and development initiatives focusing specifically on poxviruses, filoviruses, and equine encephalitis viruses (EEV). We discuss the benefits and technical challenges in the current development strategies and the hurdles in the licensure path for MCMs against these highly pathogenic viruses under the FDA Animal Rule, and we provide recommendations for the path forward.     

Transmission of aerosolized seasonal H1N1 influenza A to ferrets

Influenza virus is a major cause of morbidity and mortality worldwide, yet little quantitative understanding of transmission is available to guide evidence-based public health practice. Recent studies of influenza non-contact transmission between ferrets and guinea pigs have provided insights into the relative transmission efficiencies of pandemic and seasonal strains, but the infecting dose and subsequent contagion has not been quantified for most strains. In order to measure the aerosol infectious dose for 50% (aID₅₀) of seronegative ferrets, seasonal influenza virus was nebulized into an exposure chamber with controlled airflow limiting inhalation to airborne particles less than 5 µm diameter. Airborne virus was collected by liquid impinger and Teflon filters during nebulization of varying doses of aerosolized virus. Since culturable virus was accurately captured on filters only up to 20 minutes, airborne viral RNA collected during 1-hour exposures was quantified by two assays, a high-throughput RT-PCR/mass spectrometry assay detecting 6 genome segments (Ibis T5000™ Biosensor system) and a standard real time RT-qPCR assay. Using the more sensitive T5000 assay, the aID₅₀ for A/New Caledonia/20/99 (H1N1) was approximately 4 infectious virus particles under the exposure conditions used. Although seroconversion and sustained levels of viral RNA in upper airway secretions suggested established mucosal infection, viral cultures were almost always negative. Thus after inhalation, this seasonal H1N1 virus may replicate less efficiently than H3N2 virus after mucosal deposition and exhibit less contagion after aerosol exposure.     

A unique radiation of marine littorinid snails in the freshwater streams of the Western Ghats of India: the genus Cremnoconchus W.T. Blanford, 1869 (Gastropoda: Littorinidae)

The caenogastropod family Littorinidae is almost exclusively marine, but a unique freshwater genus, Cremnoconchus, is known from India. Its members are restricted to montane streams on the western escarpment of the Western Ghats, at altitudes between 300 and 1400 m. Four species and several varieties were described in the 19th century, but no taxonomic study has been carried out for over 120 years and the last anatomical report was in 1935. Nevertheless, they are of unusual evolutionary interest and also of conservation concern as a genus endemic to the Western Ghats biodiversity hotspot. Based on anatomical study of newly collected material and examination of historical and type specimens, we present a systematic revision of Cremnoconchus, illustrating shells, radulae, and reproductive anatomy. The very large eggs, invaginated penial filament, and calcified operculum are unique among Littorinidae. Three valid, described species (C. syhadrensis, C. conicus, C. canaliculatus) are recognized in the northern Western Ghats in Maharashtra state, where all can occasionally be found sympatrically. We describe an additional six new species from the central Western Ghats in a small area (linear distance 80 km) of Karnataka state, over 500 km south of the previously known range of the genus. Here the species each appear to be restricted to a single drainage system. Because of their highly restricted distribution and fragile habitat, this radiation of nine species is judged to be endangered. © 2013 The Linnean Society of London     

In vitro propagation and assessment of genetic stability of micropropagated Samanea saman (rain tree) using microsatellite markers

An efficient in vitro propagation of Samanea saman (rain tree) protocol has been successfully developed using nodal explants from a 20-year-old tree. Higher percentage (76 %) of explants produced up to five shoots per explant on Murashige and Skoog (MS) medium supplemented with 2 mg L^?1 6-benzyladenine (BA), 0.1 mg L^?1 gibberellic acid (GA₃) and 100 mg L^?1 casein hydrolysate after 3 weeks of culture. When explants were subcultured to fresh medium after harvesting first batch of shoots, more shoots could be generated (another eight shoots per explant). Shoot elongation was achieved (3 cm) when shoots were cultured on MS medium supplemented with 0.25 mg L^?1 BA and 0.75 mg L^?1 GA₃. In vitro generated shoots rooted on MS medium fortified with 0.75 mg L^?1 indole-3-butyric acid plus 0.1 % of activated charcoal. A higher percentage of explant response and shoots per explant were obtained on MS medium with BA and GA₃. Each responsive nodal explant yields an average of 15 rooted plants within a period of 10 weeks. Rooted plantlets were successfully acclimatized in green house with a survival rate of 90 %. Micropropagated plants were tested for genetic stability using simple sequence repeats (SSR) markers. Use of the 12 high-resolution SSR markers revealed the exact same genetic profile between the mother tree (donor) and micropropagated plants, suggesting the genetic fidelity of our micropropagation protocol. The same protocol was also used successfully in propagating a “Golden Rain Tree” although response of explant and efficiency of propagation was much lower. This protocol will be useful for germplasm preservation/large scale production of true-to-type clones of desirable genotypes.     

N‐acetylcysteine prevents oxidized low‐density lipoprotein‐induced reduction of MG53 and enhances MG53 protective effect on bone marrow stem cells

MG53 is an important membrane repair protein and partially protects bone marrow multipotent adult progenitor cells (MAPCs) against oxidized low‐density lipoprotein (ox‐LDL). The present study was to test the hypothesis that the limited protective effect of MG53 on MAPCs was due to ox‐LDL‐induced reduction of MG53. MAPCs were cultured with and without ox‐LDL (0‐20 μg/mL) for up to 48 hours with or without MG53 and antioxidant N‐acetylcysteine (NAC). Serum MG53 level was measured in ox‐LDL‐treated mice with or without NAC treatment. Ox‐LDL induced significant membrane damage and substantially impaired MAPC survival with selective inhibition of Akt phosphorylation. NAC treatment effectively prevented ox‐LDL‐induced reduction of Akt phosphorylation without protecting MAPCs against ox‐LDL. While having no effect on Akt phosphorylation, MG53 significantly decreased ox‐LDL‐induced membrane damage and partially improved the survival, proliferation and apoptosis of MAPCs in vitro. Ox‐LDL significantly decreased MG53 level in vitro and serum MG53 level in vivo without changing MG53 clearance. NAC treatment prevented ox‐LDL‐induced MG53 reduction both in vitro and in vivo. Combined NAC and MG53 treatment significantly improved MAPC survival against ox‐LDL. These data suggested that NAC enhanced the protective effect of MG53 on MAPCs against ox‐LDL through preventing ox‐LDL‐induced reduction of MG53.     

The effect of polyamines on the efficiency of multiplication and rooting of <Emphasis Type="Italic">Withania somnifera</Emphasis> (L.) Dunal and content of some withanolides in obtained plants

An efficient mass multiplication protocol was developed for Withania somnifera (L.) Dunal from nodal explants of field-grown plants on Murashige and Skoog medium (MS) supplemented with 6-benzyladenine (BA) [1.5 mg L^−l], indole-3-acetic acid (IAA) [0.3 mg L^−l] and with the addition of polyamine, spermidine (20 mg L^−l) (shoot multiplication medium). A total of 46.4 shoots were obtained from nodal explants and they were elongated in the same medium in a culture duration of 6 weeks. The elongated shoots produced roots in MS medium fortified with putrescine (20 mg L^−l) after 4 weeks, and all the rooted plants were successfully hardened and acclimatized with a survival rate of 100%. An average of 276 shoots (46 × 6) was produced when at least six nodal explants obtained from each of the 46 in vitro grown shoots were cultured by microcutting method in the same shoot multiplication medium. On an average, 12,696 plants could be produced from all the shoots (276 × 46) by microcuttings in a period of 7 months. HPLC revealed a significant increase in the quantities of withanolide A, withanolide B, withaferin A and withanone in the leaves, stems, and roots of in vitro regenerated plants compared to the field-grown parent plants. Ploidy analysis using flow cytometry revealed genetic stability of in vitro regenerated plants. This protocol will be useful for scale-up production of withanolides on commercial scale.     

Mineralization of pristine chitosan film through biomimetic process

The biomineralization of pristine chitosan film without any prior surface treatment was evaluated by immersing the film in simulated body fluid (SBF) at 37 °C for 3 weeks. The film was prepared by solvent casting method using chitosan of known degree of deacetylation (DD). The formation of the hydroxyapatite (HA) phase on the film surface after immersion was studied periodically by X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), energy dispersive X-ray analysis (EDX) and scanning electron microscopy (SEM) methods. The electron micrographs showed the morphology of the deposited apatite as small globules appearing uniformly throughout the films surfaces. The Ca/P ratio of the apatite was found to increase with increase in immersion time and approaching towards the stoichiometric value of the HA phase. The mineralized chitosan film could be of promising support to hard tissue regeneration.