Performance of scientific cameras with different sensor types in measuring dynamic processes in fluorescence microscopy

The plethora of available scientific cameras of different types challenges the biologically oriented experimenter when picking the appropriate camera for his experiment. In this study, we chose to investigate camera performances in a typical nonsingle molecule situation in life sciences, that is, quantitative measurements of fluorescence intensity changes from video data with typically skewed intensity distributions. Here, intensity profile dynamics of pH‐sensors upon triggered changes of pH‐environments in living cells served as a model system. The following camera types were tested: sCMOS, CCD (scientific and nonscientific) and EM‐CCD (back‐ and front‐illuminated). We found that although the EM‐CCD cameras achieved the best absolute spatial SNR (signal‐to‐noise ratio) values, the sCMOS was at least of equal performance when the spatial SNR was related to the effective dynamic range, and it was superior in terms of temporal SNR. In the measurements of triggered intensity changes, the sCMOS camera had the advantage that it used the smallest fraction of its dynamic range when depicting intensity changes, and thus featured the best SNR at full usage of its dynamic range. Microsc. Res. Tech. 76:835–843, 2013. © 2013 Wiley Periodicals, Inc.     

On-line recording of solar cooker use rate by a novel metering device: Prototype description and experimental verification of output data

A metering device for the determination of solar cooker use rate is presented. The device records food temperature, ambient temperature and irradiance. Automatic data evaluation yields the number of cooking cycles, cooking time, food “thermal mass”, as well as the impact on fuel consumption and GHG emission compared to other cooking techniques. Metering results are compared with actual conditions for box-type and concentrating solar cookers and found to be in agreement.     

SUCNR1 Is Expressed in Human Placenta and Mediates Angiogenesis: Significance in Gestational Diabetes

Placental hypervascularization has been reported in pregnancy-related pathologies such as gestational diabetes mellitus (GDM). Nevertheless, the underlying causes behind this abnormality are not well understood. In this study, we addressed the expression of SUCNR1 (cognate succinate receptor) in human placental endothelial cells and hypothesized that the succinate–SUCNR1 axis might play a role in the placental hypervascularization reported in GDM. We measured significantly higher succinate levels in placental tissue lysates from women with GDM relative to matched controls. In parallel, SUCNR1 protein expression was upregulated in GDM tissue lysates as well as in isolated diabetic fetoplacental arterial endothelial cells (FpECAds). A positive correlation of SUCNR1 and vascular endothelial growth factor (VEGF) protein levels in tissue lysates indicated a potential link between the succinate–SUCNR1 axis and placental angiogenesis. In our in vitro experiments, succinate prompted hallmarks of angiogenesis in human umbilical vein endothelial cells (HUVECs) such as proliferation, migration and spheroid sprouting. These results were further validated in fetoplacental arterial endothelial cells (FpECAs), where succinate induced endothelial tube formation. VEGF gene expression was increased in response to succinate in both HUVECs and FpECAs. Yet, knockdown of SUCNR1 in HUVECs led to suppression of VEGF gene expression and abrogated the migratory ability and wound healing in response to succinate. In conclusion, our data underline SUCNR1 as a promising metabolic target in human placenta and as a potential driver of enhanced placental angiogenesis in GDM.     

Combined Blockade of TIGIT and CD39 or A2AR Enhances NK-92 Cell-Mediated Cytotoxicity in AML

This study aimed to characterize different natural killer (NK) cell phenotypes on bone marrow and peripheral blood cells from acute myeloid leukemia (AML) patients and healthy donors (HDs). Our data show that CD56^dimCD16⁻ and CD56^brightCD16⁻ NK cells represent the predominant NK cell subpopulations in AML, while the CD56^dimCD16⁺ NK cells are significantly reduced compared to HDs. Moreover, TIGIT⁺ and PVRIG⁺ cells cluster on the CD56^dimCD16⁺ subset whereas CD39⁺ and CD38⁺ cells do so on CD56^brightCD16⁻ NK cells in AML. Furthermore, functional effects of (co-)blockade of TIGIT and CD39 or A2AR on NK cell functionality were analyzed. These experiments revealed that the single blockade of the TIGIT receptor results in an increased NK-92 cell-mediated killing of AML cells in vitro. Combined targeting of CD39 or A2AR significantly augments the anti-TIGIT-mediated lysis of AML cells. Our data indicate that distinct NK cell subsets in AML exhibit different immunosuppressive patterns (via the TIGIT/PVRIG receptors and the purinergic pathway). In summary, we conclude that TIGIT, CD39, and A2AR constitute relevant inhibitory checkpoints of NK cells in AML patients. A combinatorial blockade synergistically strengthens NK-92 cell-mediated cytotoxicity. As inhibitors of TIGIT, CD39, and A2AR are clinically available, studies on their combined use could be conducted in the near future.     

The Antimicrobial Peptide Gad-1 Clears Pseudomonas aeruginosa Biofilms under Cystic Fibrosis Conditions

Bacterial infections in cystic fibrosis (CF) patients are an emerging health issue and lead to a premature death. CF is a hereditary disease that creates a thick mucus in the lungs that is prone to bacterial biofilm formation, specifically Pseudomonas aeruginosa biofilms. These biofilms are very difficult to treat because many of them have antibiotic resistance that is worsened by the presence of extracellular DNA (eDNA). eDNA helps to stabilize biofilms and can bind antimicrobial compounds to lessen their effects. The metallo-antimicrobial peptide Gaduscidin-1 (Gad-1) eradicates established P. aeruginosa biofilms through a combination of modes of action that includes nuclease activity that can cleave eDNA in biofilms. In addition, Gad-1 exhibits synergistic activity when used with the antibiotics kanamycin and ciprofloxacin, thus making Gad-1 a new lead compound for the potential treatment of bacterial biofilms in CF patients.     

Improved trapped field performance of single grain Y-Ba-Cu-O bulk superconductors containing artificial holes

The intrinsic mechanical properties of single-grain RE-Ba-Cu-O bulk high-temperature superconductors can be improved by employing a thin-wall geometry. This is where the samples are melt-processed with a predefined network of artificial holes to decrease the effective wall thickness. In this study, the tensile strengths of thin-wall YBCO disks were determined using the Brazilian test at room temperature. Compared with conventional single grain YBCO disks, the thin-wall YBCO disks displayed an average tensile strength that is 93% higher when the holes were filled with Stycast epoxy resin. This implies a thin-wall sample should, in theory, be able to sustain a trapped field that is 39% higher without exceeding the mechanical limit of the sample. High-field magnetization experiments were performed by applying magnetization fields of up to 11.5 T, specifically to break the samples in order to verify the effect of increased mechanical strength (and improved cooling) on the ability of bulk (RE)BCO to trap field successfully. The standard YBCO sample failed when it was magnetized with a field of 10 T at 35 K, suffering permanent damage. As a result, the standard sample could only trap a maximum surface field of 7.6 T without failure. On the other hand, the thin-wall YBCO sample survived all magnetization cycles, including a maximum magnetization field of 11.5 T at 35 K, demonstrating a greater intrinsic ability to withstand significantly higher electromagnetic stresses. By subsequently field-cooling the thin-wall sample with 11 T at 30 K, a surface field of 8.8 T was trapped successfully without requiring any external ring reinforcement.     

Production and manipulation of elevated temperatures in pig abdomen with a three-electrode capacitive heating device

The ability of a three-electrode capacitive heating device operating at 13.56 MHz to produce and effect spatial redistribution of hyperthermic temperatures is demonstrated through abdominal heating studies conducted with 80 kg female and 65 kg male pigs. Temperature-time profiles over the respective heating periods were obtained in anterior and posterior superficial tissue and in selected abdominal organs. At selected times during heating, spatial redistribution of elevated temperatures was effected through adjustment of power settings for one or more of the three electrodes, i.e. through manipulation of the specific absorption rate (SAR) in-vivo.     

Activation of CNS circuits producing a neurogenic cystitis: evidence for centrally induced peripheral inflammation

We present a model of neurogenic cystitis induced by viral infection of specific neuronal circuits of the rat CNS. Retrograde infection by pseudorabies virus (PRV) of neuronal populations neighboring those that innervate the bladder consistently led to a localized immune response in the CNS and bladder inflammation. Infection of bladder circuits themselves or of circuits distant from these rarely produced cystitis. Absence of virus in bladder and urine ruled out an infectious cystitis. Total denervation of the bladder, selective C-fiber deafferentation, or bladder sympathectomy prevented cystitis without affecting the CNS disease, indicating a neurogenic component to the inflammation. The integrity of central bladder-related circuits is necessary for the appearance of bladder inflammation, because only CNS lesions affecting bladder circuits, i.e., bilateral dorsolateral or ventrolateral funiculectomy, as well as bilateral lesions of Barrington's nucleus/locus coeruleus area, prevented bladder inflammation. The close proximity in the CNS of noninfected visceral circuits to infected somatic neurons would thus permit a bystander effect, leading to activation of the sensory and autonomic circuits innervating the bladder and resulting in a neurogenic inflammation localized to the bladder. The present study indicates that CNS dysfunction can bring about a peripheral inflammation.