Remote optogenetic control of the enteric nervous system and brain-gut axis in freely-behaving mice enabled by a wireless, battery-free optoelectronic device.

Wireless activation of the enteric nervous system (ENS) in freely moving animals with implantable optogenetic devices offers a unique and exciting opportunity to selectively control gastrointestinal (GI) transit in vivo, including the gut-brain axis. Programmed delivery of light to targeted locations in the GI-tract, however, poses many challenges not encountered within the central nervous system (CNS). We report here the development of a fully implantable, battery-free wireless device specifically designed for optogenetic control of the GI-tract, capable of generating sufficient light over large areas to robustly activate the ENS, potently inducing colonic motility ex vivo and increased propulsion in vivo. Use in in vivo studies reveals unique stimulation patterns that increase expulsion of colonic content, likely mediated in part by activation of an extrinsic brain-gut motor pathway, via pelvic nerves. This technology overcomes major limitations of conventional wireless optogenetic hardware designed for the CNS, providing targeted control of specific neurochemical classes of neurons in the ENS and brain-gut axis, for direct modulation of GI-transit and associated behaviours in freely moving animals.

Effects of optogenetic activation of the enteric nervous system on gastrointestinal motility in mouse small intestine.

BACKGROUND AND AIMS: Recently, it was demonstrated that optogenetics could be used to stimulate enteric calretinin neurons, leading to increased colonic transit in vitro and in vivo. The aim of the current study was to determine if similar approaches could be used to stimulate the isolated mouse small intestine, with the aim of potentially also improving transit in the small bowel. METHODS: Cre-Lox recombination was used to generate transgenic mice expressing the light-sensitive ion channel channelrhodopsin-2 (ChR2) in calretinin neurons. RESULTS: Spontaneous migrating motor complexes were recorded from isolated terminal small intestine in both CalCre+ mice expressing ChR2 in calretinin-expressing neurons and experimental controls, CalCre-. Trains of blue light pulses (20 ms, 5 Hz, 20s) evoked a brief local contraction of circular muscle, but never a premature MMC, irrespective of light intensity (1-40 mV/mm2) or the region of ileum stimulated. However, MMCs were readily evoked by calretinin neuron activation in colon, consistent with our previous study. Light-evoked contractions of the terminal small bowel were hexamethonium-resistant (300 µM), but blocked by tetrodotoxin (0.6 µM). Light-evoked smooth muscle contraction did not change the pacemaker frequency underlying MMCs. CONCLUSION: Focal illumination of the small intestine does not appear as effective at inducing propulsive motor activity as has been demonstrated in the colon of the same colony mice. This study suggests caution should be exercised when assuming optogenetic technology is equally effective at increasing GI transit in the small intestine as in the large intestine of mice.

Photoluminescence excited at variable fluences: a novel approach for studying the emission from crystalline pigments in paints.

Crystalline solids can exhibit photoluminescence when properly excited by sufficiently energetic light radiation. Following excitation, different radiative and non-radiative recombination pathways can occur that are informative of the energetic structure of the material as well as of the presence of crystal defects and impurities. Usually, the characterization of the optical emission of crystalline materials is achieved through the study of emission spectra as a function of the excitation wavelength. A different approach employs variable excitation fluence to populate the energetic levels until saturation, which promotes the emission from other radiative and non-radiative pathways. The method is particularly effective for understanding conduction phenomena and studying charge recombination channels in semiconductor materials. In this work, we propose its application for characterizing radiative recombination paths in crystalline pigments. The approach has been tested in spectroscopy mode for the identification of paints in a model painting and in micro-imaging modality for the study of paint stratigraphies. We demonstrate that the method is highly informative of the nature of different recombination paths in crystalline pigments and allows a deeper characterization of the emission from luminescent paints with respect to the conventional steady-state photoluminescence approach.

High temperature, transparent, superhydrophobic Teflon AF-2400/Indium tin oxide nanocomposite thin films.

The outstanding properties of Teflon AF-2400-chemical, optical, etc-inspired us to make modifications to enhance its hydrophobicity. We prepared an AF-2400/indium tin oxide (ITO) nanocomposite by a spin coating technique at room temperature, using the AF-2400 polymer as the matrix and ITO nanoparticles as the filler. Different ITON concentrations ranging from 3 to 30 mg ml-1 were prepared to study the effect of nanoparticle loading on the films' properties and superhydrophobicity. The effect of spin speed and annealing temperature was also studied. Atomic force microscopy, x-ray photoelectron spectroscopy, and UV-vis analysis were employed to characterize the prepared films. The results indicated that the film's low surface energy and nano/micro-features made it superhydrophobic. Increasing the ITON concentration to 15 mg ml-1 improved the superhydrophobicity of the composite film by increasing the surface roughness. The coating showed superhydrophobic behavior with a static contact angle (SCA) around 152° and contact angle hysteresis less than 2°. The nanocomposite films also exhibited excellent thermal stability, sustaining temperatures as high as 240 °C without losing their superhydrophobic behavior. Three models, Wenzel, Cassie-Baxter, and Shuttleworth-Bailey, were used to predict the SCA. The results confirmed that the latter model gave the best prediction. In addition to superhydrophobicity, the AF-2400/ITON films coated on a glass substrate showed very high transparency-around 95% in the visible and infrared ranges. An effective medium theory, the Bergman representation, was used to simulate the transmittance of the AF-2400/ITON nanocomposites. The measured and simulated transmittance values were in good agreement in the visible range. Based on our results, this coating may be highly useful for many practical applications, including solar cell coatings, chemical resistance protective coatings, and more.

Antimicrobial resistance in group B streptococcus: the Australian experience.

Intrapartum chemoprophylaxis for pregnant group B streptococcus (GBS) carriers reduces vertical transmission, with a resultant decrease in neonatal as well as maternal morbidity from invasive GBS infection. Current Australian guidelines recommend penicillin for intrapartum prophylaxis of GBS carriers, with erythromycin or clindamycin for those with a ß-lactam allergy. Recent reports globally suggest that resistance to erythromycin and clindamycin may be increasing; hence, a study was undertaken to promote an evidence base for local clinical guidelines. Samples collected for standardized susceptibility testing included 1160 invasive GBS isolates (264 isolates retrospectively from 1982 to 2001 and prospectively from 2002 to 2006, plus 896 prospectively collected colonizing GBS isolates gathered over a 12 month period from 2005 to 2006) from 16 laboratories around Australia. All isolates displaying phenotypic macrolide or lincosamide resistance were subsequently genotyped. No isolates showed reduced susceptibility to penicillin or vancomycin. Of the invasive isolates, 6.4 % demonstrated phenotypic erythromycin resistance and 4.2 % were clindamycin resistant. Of the erythromycin-resistant isolates, 53 % showed cross-resistance to clindamycin. Very similar results were found in colonizing specimens. There was no statistically significant change in macrolide-resistance rates over the two study periods 1982-2001 and 2002-2006. Genotyping for macrolide and lincosamide-resistant isolates was largely consistent with phenotype. These findings suggest that penicillin therapy remains an appropriate first-line antibiotic choice for intrapartum GBS chemoprophylaxis, with erythromycin and/or clindamycin resistance being low in the Australian population. It would, nevertheless, be appropriate for laboratories screening for GBS in obstetric patients to consider macrolide sensitivity testing, particularly for those with ß-lactam allergy, to ensure appropriate chemoprophylaxis.

Screening protocols for group B streptococcus: are transport media appropriate?

OBJECTIVE: To evaluate group B streptococcus (GBS) detection in an in vitro setting, using a low and controlled inoculum from swabs directly inoculated into a selective medium, as compared to delayed inoculation following a period in a commercial Amies transport medium with charcoal (Venturi Transystem Copan, Italy). STUDY DESIGN: Clinical isolates of GBS (n = 103), were inoculated into the Amies transport medium with charcoal in a concentration of 100 colony-forming units (cfu)/ml (10 cfu/swab). Swabs were then transferred to an enrichment broth (NPC) at time intervals of 0, 2, 4, 6 and 24 hours. Broths were then incubated for 18-24 hours at 35 degrees C in air, before being transferred to New Granada Medium Modified (NGM) for GBS detection and incubated for a further 18-24 hours at 35 degrees C in air. If the characteristic orange pigmented colonies were observed after this period, the specimen was recorded as + (1-10 colonies) or + + (more than 10 colonies). RESULTS: Overall 92.2% (95/103) of isolates were detected in all tubes and at all times. An additional two isolates were non-hemolytic, non-pigment forming GBS. Of note, 3.9% (4/103) were negative until 2 hours delayed inoculation and 1.9% (2/103) gave inconsistent results, likely due to the low inoculum used. CONCLUSION: Delayed inoculation into selective enrichment broth following a period in transport medium, even with a low inoculum, gave a similar and acceptable GBS detection rate to direct inoculation. Hence, Amies transport medium with charcoal is an appropriate transport medium to use, where it is not practical for clinical specimens to be directly inoculated into selective enrichment broth and as endorsed in the Centers for Diseases Control (CDC) Guidelines, 2002.

Is penicillin and/or erythromycin resistance present in clinical isolates of group B streptococcus in our community?

Traditionally group B streptococcus (GBS) has not exhibited resistance to the antibiotics penicillin and erythromycin. Recently there are reports from North America whereby some GBS isolates have exhibited resistance to erythromycin. There have been no studies reported to date to determine whether this resistance trend is also occurring in the Australian population. Over a period of six months from January to June 1999, 250 GBS isolates were collected at the Royal Women's Hospital, Melbourne. Sensitivity to penicillin, erythromycin and vancomycin was determined by disk diffusion. Any isolates that had reduced zones to penicillin, vancomycin or erythromycin had minimum inhibitory concentrations (MIC) determined. No isolates had reduced susceptibility to penicillin or vancomycin. Of the 250 isolates, seven (2.8%, 95% CI = 1.1-5.6%) had resistant MICs to erythromycin of > 1.0 microg/mL. These preliminary data suggest that penicillin therapy is still an effective first-line antibiotic for intrapartum chemoprophylaxis and that erythromycin resistance is low in our population.

Clinical implications of inducible beta-lactamase activity in Gram-negative bacteremia in children.

BACKGROUND: Organisms of the spp., indole-positive spp., spp. and (ESCaPPM) group are a common cause of hospital-acquired bacteremia and share the potential to develop beta-lactam resistance during therapy. The emergence of such resistance may have adverse consequences, but the frequency with which this occurs has not been studied in children. It has been suggested that such organisms should be treated with combination antimicrobials or carbapenems, but the optimal regimen is uncertain. AIM To determine the frequency with which beta-lactam resistance develops during ESCaPPM sepsis in children and the optimal treatment of such sepsis. METHODS: A review of the case notes and microbiologic records of all cases of ESCaPPM bacteremia and meningitis managed at a tertiary children's hospital during a 6-year period. RESULTS: Fifty-eight cases were identified, and in three (5%) cases beta-lactam resistance emerged during treatment, with adverse clinical consequences in two cases. Clinical and microbiologic outcome was similar in those treated with carbapenems and in those treated with a beta-lactam and aminoglycoside combination. Cefotaxime resistance was found in 57, 30, 24 and 7% of children who had received carbapenems, cephalosporins, penicillins or no/other antimicrobials in the month before ESCaPPM sepsis, respectively. CONCLUSIONS: The emergence of beta-lactam resistance during treatment of ESCaPPM sepsis is uncommon in our hospital but can have adverse consequences. Where isolates are reported as susceptible to both classes of drugs, an extended spectrum penicillin in combination with an aminoglycoside may be preferable first line treatment of ESCaPPM sepsis to a carbapenem or quinolone.