Luminal Space Research Articles

Improved background modeling of video sequences using spatio-temporal extension of fuzzy local binary pattern

Background subtraction is a method of motion detection in video sequences captured by static camera based on construction of a background model and its progressive comparison with each frame of the video. Sometimes the changes in the background objects are not permanent and appear at a rate faster than that of the background update, and this leads to emergence of dynamic textures in the background. As a result, high-quality background modeling plays a major role in motion detection performance, especially in videos with dynamic backgrounds and adverse environmental conditions such as noise. Although Local Binary Pattern (LBP) is a successful methodology for background subtraction, but it cannot properly extract textures from uniform areas of the foreground. In recent years, Fuzzy Local Binary Pattern (FLBP) has been developed to improve the performance of LBP operator in texture extraction from images with additive noise. The use of FLBP texture descriptor for background subtraction has led to improved robustness against noise and low sensitivity of the background model to slight changes in the texture gray scale values, and therefore better texture extraction from uniform areas, even in the presence of dynamic backgrounds and adverse environmental conditions. But despite this improvement, this operator is still sensitive to time-variations of pixels in dynamic backgrounds. To avoid this issue and incorporate correlation of pixel values over successive frames, this study proposes the use of spatio-temporal extension of FLBP with symmetry about central pixel in combination with the Local Color Histogram (LCH) in the Improved Hue Luminance and Saturation (IHLS) color space for describing the pixel color features. The results of tests conducted with standard databases show that for dynamic backgrounds, the use of proposed Spatio-Temporal Fuzzy Center Symmetric Local Binary Pattern (STFCS-LBP) operator with the spatio-temporal neighborhood texture patterns and the local color histogram yields better results than the existing methods.

"Atrophic Kidney"-like Lesion: Clinicopathologic Series of 8 Cases Supporting a Benign Entity Distinct From Thyroid-like Follicular Carcinoma.

Renal mass lesions with a follicular architecture resembling atrophic kidney have been described, but their distinction from thyroid-like follicular carcinoma of the kidney remains controversial. We collected 8 cases of this purported "atrophic kidney"-like lesion to fully describe their clinical and histologic spectrum, their possible etiology, and to discuss their distinction from other renal neoplasms. Eight total cases were identified with patient ages ranging from 9 to 48 years (mean: 29 y; median: 28.5 y). Four patients were female and 4 were male. The tumors were unifocal and size ranged from 1.6 to 4.9 cm (mean: 3.4 cm; median: 3.4 cm). All 8 tumors had a remarkably similar histology. Each was enveloped by a smooth muscle rich capsule and had an overall low power "follicular" architecture. The luminal spaces of the "follicles" (or cysts) contained eosinophilic secretions and the lining epithelium was often flattened and atrophic, but some had more rounded cells with a distinctive hobnail arrangement. Many cysts contained discohesive round cells floating within the eosinophilic material, and some contained small intraluminal tufts with features of markedly atrophic glomeruli. Periodic acid-Schiff stains highlighted basement membrane material extending into these glomerular-like tufts, and some contained small distinct capillaries surrounded by endothelial cells, interspersed mesangial-like cells, and rare surrounding podocyte-like cells, providing additional evidence for glomerulocystic structures. Scattered calcifications were present within cysts (or within cyst walls) in varying numbers and were characterized by 2 types: psammoma body-like or more amorphous deposits. The tissue between cystic glomeruli contained predominantly small atrophic tubular structures, but collagenized stroma and smaller collapsed glomeruli were also present. The 2 tumors from the oldest 2 patients (48 and 39 y) had a more striking degree of stromal hyalinization. Immunohistochemically, the cyst lining cells had a predominant WT-positive/PAX-8 negative/CK7-negative phenotype, while tubules were typically WT-1 negative/PAX-8 positive/CK7-positive. Upon comparison to a control group of 10 kidneys containing incidental non-mass-forming glomerulocystic change, the morphologic features and immunophenotype were identical. To date, no patient has had any recurrence or aggressive clinical behavior based on follow status in 7 of 8 cases (follow-up range: 9 to 168 mo; median: 24 mo; mean: 40 mo). In summary, we describe the clinicopathologic features of 8 unique, benign "atrophic kidney"-like lesions that may simply represent a non-neoplastic form of organizing tubular atrophy and glomerulocystic change, and emphasize their distinction from thyroid-like follicular carcinoma of the kidney.

Symposium review: Intravital imaging of the lactating mammary gland in live mice reveals novel aspects of milk-lipid secretion

Milk fat comprises membrane-coated droplets of neutral lipid, which constitute the predominant source of lipids for survival of the suckling neonate. From the perspective of the dairy industry, they are the basis for the manufacture of butter and essential ingredients in the production of cheese, yogurt, and specialty dairy produce. To provide mechanistic insight into the assembly and secretion of lipid droplets during lactation, we developed novel intravital imaging techniques using transgenic mice, which express fluorescently tagged marker proteins. The number 4 mammary glands were surgically prepared under a deep plane of anesthesia and the exposed glands positioned as a skin flap with intact vascular supply on the stage of a laser-scanning confocal microscope. Lipid droplets were stained by prior exposure of the glands to hydrophobic fluorescent BODIPY (boron-dipyrromethene) dyes and their formation and secretion monitored by time-lapse subcellular microscopy over periods of 1 to 2 h. Droplets were transported to the cell apex by directed (superdiffusive) motion at relatively slow and intermittent rates (0–2 µm/min). Regardless of size, droplets grew by numerous fusion events during transport and as they were budding from the cell enveloped by apical membranes. Surprisingly, droplet secretion was not constitutive but required an injection of oxytocin to induce contraction of the myoepithelium with subsequent release of droplets into luminal spaces. These novel results are discussed in the context of the current paradigm for milk fat synthesis and secretion and as a template for future innovations in the dairy industry.

A Review: The Fate of Bacteriocins in the Human Gastro-Intestinal Tract: Do They Cross the Gut-Blood Barrier?

The intestinal barrier, consisting of the vascular endothelium, epithelial cell lining, and mucus layer, covers a surface of about 400 m2. The integrity of the gut wall is sustained by transcellular proteins forming tight junctions between the epithelial cells. Protected by three layers of mucin, the gut wall forms a non-permeable barrier, keeping digestive enzymes and microorganisms within the luminal space, separate from the blood stream. Microorganisms colonizing the gut may produce bacteriocins in an attempt to outcompete pathogens. Production of bacteriocins in a harsh and complex environment such as the gastro-intestinal tract (GIT) may be below minimal inhibitory concentration (MIC) levels. At such low levels, the stability of bacteriocins may be compromised. Despite this, most bacteria in the gut have the ability to produce bacteriocins, distributed throughout the GIT. With most antimicrobial studies being performed in vitro, we know little about the migration of bacteriocins across epithelial barriers. The behavior of bacteriocins in the GIT is studied ex vivo, using models, flow cells, or membranes resembling the gut wall. Furthermore, little is known about the effect bacteriocins have on the immune system. It is generally believed that the peptides will be destroyed by macrophages once they cross the gut wall. Studies done on the survival of neurotherapeutic peptides and their crossing of the brain–blood barrier, along with other studies on small peptides intravenously injected, may provide some answers. In this review, the stability of bacteriocins in the GIT, their effect on gut epithelial cells, and their ability to cross epithelial cells are discussed. These are important questions to address in the light of recent papers advocating the use of bacteriocins as possible alternatives to, or used in combination with, antibiotics.

IRTKS (BAIAP2L1) Elongates Epithelial Microvilli Using EPS8-Dependent and Independent Mechanisms

Transporting epithelial cells like those that line the gut build large arrays of actin-supported protrusions called microvilli, which extend from the apical surface into luminal spaces to increase functional surface area. Although critical for maintaining physiological homeostasis, mechanisms controlling the formation of microvilli remain poorly understood. Here, we report that the inverse-bin-amphiphysin-Rvs (I-BAR)-domain-containing protein insulin receptor tyrosine kinase substrate (IRTKS) (also known as BAIAP2L1) promotes the growth of epithelial microvilli. Super-resolution microscopy and live imaging of differentiating epithelial cells revealed that IRTKS localizes to the distal tips of actively growing microvilli via a mechanism that requires its N-terminal I-BAR domain. At microvillar tips, IRTKS promotes elongation through a mechanism involving its C-terminal actin-binding WH2 domain. IRTKS can also drive microvillar elongation using its SH3 domain to recruit the bundling protein EPS8 to microvillar tips. These results provide new insight on mechanisms that control microvillar growth during the differentiation of transporting epithelial cells and help explain why IRTKS is targeted by enteric pathogens that disrupt microvillar structure during infection of the intestinal epithelium.

In vivo imaging reveals unique neutrophil transendothelial migration patterns in inflamed intestines.

Neutrophil (PMN) infiltration of the intestinal mucosa is a hallmark of gastrointestinal inflammation, with significant implications for host defense, injury and repair. However, phenotypic and mechanistic aspects of PMN recruitment in inflamed intestines have not been explored in vivo. Using novel epithelial/PMN fluorescence reporter mice, advanced intravital imaging and 3D reconstruction analysis, we mapped the microvasculature architecture across the intestinal layers and determined that in response to Sa/mone//a/endotoxin-induced inflammation, PMN transendothelial migration (TEM) was restricted to submucosal vessels. PMN TEM was not observed in villus or crypt vessels, proximal to the epithelium that underlies the intestinal lumen, and was partially dependent on (C-X-C motif) ligands 1 (CXCL1) and 2 (CXCL2) expression, which was found to be elevated in the submucosa layer. Restricted PMN extravasation at the submucosa and subsequent PMN interstitial migration may serve as a novel regulatory step of PMN effector function and recruitment to the luminal space in inflamed intestines.

Renal immune surveillance and dipeptidase-1 contribute to contrast-induced acute kidney injury.

Radiographic contrast agents cause acute kidney injury (AKI), yet the underlying pathogenesis is poorly understood. Nod-like receptor pyrin containing 3-deficient (Nlrp3-deficient) mice displayed reduced epithelial cell injury and inflammation in the kidney in a model of contrast-induced AKI (CI-AKI). Unexpectedly, contrast agents directly induced tubular epithelial cell death in vitro that was not dependent on Nlrp3. Rather, contrast agents activated the canonical Nlrp3 inflammasome in macrophages. Intravital microscopy revealed diatrizoate (DTA) uptake within minutes in perivascular CX3CR1+ resident phagocytes in the kidney. Following rapid filtration into the tubular luminal space, DTA was reabsorbed and concentrated in tubular epithelial cells via the brush border enzyme dipeptidase-1 in volume-depleted but not euvolemic mice. LysM-GFP+ macrophages recruited to the kidney interstitial space ingested contrast material transported from the urine via direct interactions with tubules. CI-AKI was dependent on resident renal phagocytes, IL-1, leukocyte recruitment, and dipeptidase-1. Levels of the inflammasome-related urinary biomarkers IL-18 and caspase-1 were increased immediately following contrast administration in patients undergoing coronary angiography, consistent with the acute renal effects observed in mice. Taken together, these data show that CI-AKI is a multistep process that involves immune surveillance by resident and infiltrating renal phagocytes, Nlrp3-dependent inflammation, and the tubular reabsorption of contrast via dipeptidase-1.

Pathology after Chlamydia trachomatis infection is driven by nonprotective immune cells that are distinct from protective populations

Infection with Chlamydia trachomatis drives severe mucosal immunopathology; however, the immune responses that are required for mediating pathology vs. protection are not well understood. Here, we employed a mouse model to identify immune responses required for C. trachomatis-induced upper genital tract pathology and to determine whether these responses are also required for bacterial clearance. In mice as in humans, immunopathology was characterized by extravasation of leukocytes into the upper genital tract that occluded luminal spaces in the uterus and ovaries. Flow cytometry identified these cells as neutrophils at early time points and CD4+ and CD8+ T cells at later time points. To determine what draws these cells to C. trachomatis-infected tissue, we measured the expression of 700 inflammation-related genes in the upper genital tract and found an up-regulation of many chemokines, including a node of interaction between CXCL9/10/11 and their common receptor CXCR3. Either depleting neutrophils or reducing T-cell numbers by CXCR3 blockade was sufficient to significantly ameliorate immunopathology but had no effect on bacterial burden, demonstrating that these responses are necessary for mucosal pathology but dispensable for C. trachomatis clearance. Therapies that specifically target these host responses may therefore prove useful in ameliorating C. trachomatis-induced pathology without exacerbating infection or transmission.

Acidic and Basic pH Effect in Two Cytoplasmic and Endoplasmic Reticulum Luminal Spaces on Chloride Channel Electrophysiological Behavior

Background: In regard of chloride channel electrophysiological behavior importance in cellular homeostasis maintenance, some of diseases appearance because of chloride channels impairment, also reports of synchronization between chloride channels impairment and misadjusted pH and that presumably acid or basic pH in cytoplasmic and endoplasmic reticulum luminal spaces are effective on this behavior, current study was performed. M aterials and Methods: Research was performed by experimental method. Vesicles from rat liver tissue endoplasmic reticulum were extracted and assessed in 30 samples in 6 groups. Electrophysiological behaviors of channels were measured in control, acidic and basic pH in cis and Trans environments and according of channel conductance and Po this behavior was determined and judged statistically. Data were filtered at 1 kHz and stored at a sampling rate of 10 kHz for offline analysis by PClamp9. Statistical analysis was performed based on Markov noise free single channel analysis. R es ults: Channel conductance was 72 pS and its current – Voltage relation curve was linear. Channel has Voltage dependent behavior and has grater Po in positive Voltages. Channel conductance in acidic pH remained at 72 pS as of control situation. Channel Po was not changed. In basic pH these findings were also repeated. Also, in cis and Trans spaces these behaviors were sawed. C onclusion: It seems that in pH stream from 6 to 8.5, current channel electrophysiological behavior could be important in endoplasmic reticulum and cellular homeostasis maintenance especially in positive ion such as calcium ion accumulation situation in cytoplasm.

Isolation and Culture of Juvenile Pig Thyroid Follicular Epithelia.

Epithelial tissues are defined by their polarity and their ability to transport directionally. Thyroid is a tissue comprising functional epithelial units organized as enclosed follicles, with their luminal spaces defined by thyrocyte apices. Thus, the native arrangement of thyroid epithelia limits accessibility to the follicular space, presenting a challenge in studying transepithelial movements. This limitation can be overcome by studying thyrocytes grown as two-dimensional cultures. Herein we present methods for isolation of thyroid follicles from juvenile pigs and preparation of high-resistance, polarized cultures.

A Mathematical Model Supports a Key Role for Ae4 (Slc4a9) in Salivary Gland Secretion.

We develop a mathematical model of a salivary gland acinar cell with the objective of investigating the role of two [Formula: see text] exchangers from the solute carrier family 4 (Slc4), Ae2 (Slc4a2) and Ae4 (Slc4a9), in fluid secretion. Water transport in this type of cell is predominantly driven by [Formula: see text] movement. Here, a basolateral [Formula: see text] adenosine triphosphatase pump (NaK-ATPase) and a [Formula: see text]-[Formula: see text]-[Formula: see text] cotransporter (Nkcc1) are primarily responsible for concentrating the intracellular space with [Formula: see text] well above its equilibrium potential. Gustatory and olfactory stimuli induce the release of [Formula: see text] ions from the internal stores of acinar cells, which triggers saliva secretion. [Formula: see text]-dependent [Formula: see text] and [Formula: see text] channels promote ion secretion into the luminal space, thus creating an osmotic gradient that promotes water movement in the secretory direction. The current model for saliva secretion proposes that [Formula: see text] anion exchangers (Ae), coupled with a basolateral [Formula: see text] ([Formula: see text]) (Nhe1) antiporter, regulate intracellular pH and act as a secondary [Formula: see text] uptake mechanism (Nauntofte in Am J Physiol Gastrointest Liver Physiol 263(6):G823-G837, 1992; Melvin et al. in Annu Rev Physiol 67:445-469, 2005. https://doi.org/10.1146/annurev.physiol.67.041703.084745 ). Recent studies demonstrated that Ae4 deficient mice exhibit an approximate [Formula: see text] decrease in gland salivation (Peña-Münzenmayer et al. in J Biol Chem 290(17):10677-10688, 2015). Surprisingly, the same study revealed that absence of Ae2 does not impair salivation, as previously suggested. These results seem to indicate that the Ae4 may be responsible for the majority of the secondary [Formula: see text] uptake and thus a key mechanism for saliva secretion. Here, by using 'in-silico' Ae2 and Ae4 knockout simulations, we produced mathematical support for such controversial findings. Our results suggest that the exchanger's cotransport of monovalent cations is likely to be important in establishing the osmotic gradient necessary for optimal transepithelial fluid movement.

Evidence for protandry in Polydactylus quadrifilis in the Kwanza Estuary, Angola, and its implications for local fisheries

A total of 141 Polydactylus quadrifilis were sampled from the Kwanza Estuary in Angola ranging in size from 436 to 1360 mm fork-length (FL). Of these, 124 were male, six intersex and 11 female. Female fish were significantly longer (mm, FL) and heavier (kg) than males and had significantly higher gonadosomatic indices (GSI’s) than those of males and intersex fish. Transitional (intersex) gonads were delimited, with testicular and ovarian regions separated by connective tissue. The first signs of ovarian tissue appeared on the outer ventral surface of the gonad. A second layer of ovarian tissue was first noticeable at either end of the initial ovarian region and developed back towards the centre of the ventral wall to form a luminal space. Early-stage oocytes were commonly found in the outer area of male regions and residual late-stage spermatids and spermatozoa were found in the luminal space of ovarian regions, suggesting a process of sex change from the outside inwards. A loss of male function was noted with increased ovarian prevalence. Based on this evidence it is suggested that P. quadrifilis in the Kwanza Estuary are protandrous. Owing to the reliance of P. quadrifilis on large highly fecund females for egg production, it is likely that they will be sensitive to fishing practices that target larger individuals within the population.

Non-Invasive Prostate Cancer Characterization with Diffusion-Weighted MRI: Insight from In silico Studies of a Transgenic Mouse Model.

Diffusion-weighted magnetic resonance imaging (DWI) enables non-invasive, quantitative staging of prostate cancer via measurement of the apparent diffusion coefficient (ADC) of water within tissues. In cancer, more advanced disease is often characterized by higher cellular density (cellularity), which is generally accepted to correspond to a lower measured ADC. A quantitative relationship between tissue structure and in vivo measurements of ADC has yet to be determined for prostate cancer. In this study, we establish a theoretical framework for relating ADC measurements with tissue cellularity and the proportion of space occupied by prostate lumina, both of which are estimated through automatic image processing of whole-slide digital histology samples taken from a cohort of six healthy mice and nine transgenic adenocarcinoma of the mouse prostate (TRAMP) mice. We demonstrate that a significant inverse relationship exists between ADC and tissue cellularity that is well characterized by our model, and that a decrease of the luminal space within the prostate is associated with a decrease in ADC and more aggressive tumor subtype. The parameters estimated from our model in this mouse cohort predict the diffusion coefficient of water within the prostate-tissue to be 2.18 × 10−3 mm2/s (95% CI: 1.90, 2.55). This value is significantly lower than the diffusion coefficient of free water at body temperature suggesting that the presence of organelles and macromolecules within tissues can drastically hinder the random motion of water molecules within prostate tissue. We validate the assumptions made by our model using novel in silico analysis of whole-slide histology to provide the simulated ADC (sADC); this is demonstrated to have a significant positive correlation with in vivo measured ADC (r2 = 0.55) in our mouse population. The estimation of the structural properties of prostate tissue is vital for predicting and staging cancer aggressiveness, but prostate tissue biopsies are painful, invasive, and are prone to complications such as sepsis. The developments made in this study provide the possibility of estimating the structural properties of prostate tissue via non-invasive virtual biopsies from MRI, minimizing the need for multiple tissue biopsies and allowing sequential measurements to be made for prostate cancer monitoring.

AMELIORATIVE EFFECTS OF PHYTOANDROGEN AND ANTIOXIDANT ON HISTOARCHITECTURE OF PERMETHRIN INDUCED CHANGES IN GOAT TESTIS

Objective: Protective effects of Tribulus terrestris (TT) and vitamin C were studied against permethrin induced changes in the histoarchitecture of goat testis.Methods: Hydroalcoholic extract of the fruits of Tribulus terrestris was prepared by maceration method and the extractive value was calculated. Fresh stock solution of permethrin (1mg/ml) in dimethyl sulphoxide and Tribulus terrestris (1mg/ml) in distilled water were prepared. Testicular tissues were cultured in TCM-199 nutrition media at 39℃ with 95% humidity and 5% CO₂ in the CO₂ incubator. Cultured testicular tissue was grouped as control having only the nutritive media, experimental group I(EI) received two doses of permethrin (100µg/ml and 200µg/ml), experimental group II(EII) was administered doses of permethrin along with vitamin C(0.1µM/ml) and experimental group III(EIII) was supplemented with doses of permethrin along with Tribulus terrestris (100µg/ml). All the groups were tested for 4 hr. and 8hr. for histoarcitectural studies.Results: Histological alterations were observed such as distorted shape of seminiferous tubules, decreased luminal and interstitial space, reduction in the population of spermatocytes, spermatozoa and spermatids in treated (E1) group as compared to control group. Maximum damage was detected at a higher dose of permethrin (200µg/ml) for 8h in contrast to lower doses (100 4h and 8h). Thus, it is concluded that permethrin has both dose and time-dependent effects. Protective groups (E2 and E3) imparted ameliorative effects by reestablishing shape of seminiferous tubules, decreasing luminal and interstitial space, increased population of spermatocyte, spermatozoa, and spermatids as compared with treated groups (E1)Conclusion: It has been reported that permethrin affects male fertility by inducing detrimental changes in the structure and function of seminiferous tubules. Both vitamin C and TT have the ability to mitigate the effect of pesticide. TT shows aphrodisiac activity against permethrin induced alterations in goat testis.

Verifying the Images Authenticity in Cognitive Internet of Things (CIoT)-Oriented Cyber Physical System

With the recent development of Cognitive Internet of Things (CIoT) and the potential of Cyber Physical System (CPS), people’s daily activities become smarter, and intelligent. The combination of CIoT and CPS can greatly enhance the quality of people’s life. To this end, this article proposes CIoT-CPS that comprises of two main models: user activity cognitive model and image authentication model.The user activity cognitive model (UACM) is a machine-learning model to have the meaningful data. The image authentication model is to verify the authenticity of images captured by various devices, such as smart phones, digital cameras, and other camera-embedded portable devices. The authenticity of an image is breached when parts of images are assembled to produce a new image (known as a splicing forgery), or a part of an image is copied or pasted into another part of the same image (known as a copy-move forgery). In the proposed verification method, an opposite color local binary pattern (OC-LBP) texture descriptor is applied to a questioned image. The image is first decomposed into an RGB (red, green, blue) and a luminance and chroma color spaces. The OC-LBP measures the interrelation between pixels of different color components. The intensive computation involving six color components and a gray version is performed in the cloud, where a server can be dedicated to doing this job. The histograms of the OC-LBP are concatenated with weights to produce a final feature vector of the image. A support vector machine is applied as a classifier, which classifies the image as authentic or forged. Several experiments were performed to verify the suitability of those models or approaches. The proposed approaches show a good accuracy compared to other competing approaches.

Development of hepatoma-derived, bidirectional oval-like cells as a model to study host interactions with hepatitis C virus during differentiation.

Directed differentiation of human stem cells including induced pluripotent stem cells into hepatic cells potentially leads to acquired susceptibility to hepatitis C virus (HCV) infection. However, cellular determinants that change their expression during cell reprogramming or hepatic differentiation and are pivotal for supporting the HCV life cycle remain unclear. In this study, by introducing a set of reprogramming factors, we established HuH-7-derived oval-like cell lines, Hdo-17 and -23, which possess features of bipotential liver precursors. Upon induction of hepatocyte differentiation, expression of mature hepatocyte markers and hepatoblast markers in cells increased and decreased, respectively. In contrast, in response to cholangiocytic differentiation induction, gene expression of epithelium markers increased and cells formed round cysts with a central luminal space. Hdo cells lost their susceptibility to HCV infection and viral RNA replication. Hepatic differentiation of Hdo cells potentially led to recovery of permissiveness to HCV RNA replication. Gene expression profiling showed that most host-cell factors known to be involved in the HCV life cycle, except CD81, are expressed in Hdo cells comparable to HuH-7 cells. HCV pseudoparticle infectivity was significantly but partially recovered by ectopic expression of CD81, suggesting possible involvement of additional unidentified factors in HCV entry. In addition, we identified miR200a-3p, which is highly expressed in Hdo cells and stem cells but poorly expressed in differentiated cells and mature hepatocytes, as a novel negative regulator of HCV replication. In conclusion, our results showed that epigenetic reprogramming of human hepatoma cells potentially changes their permissivity to HCV.

Abstract 5478: The TRPA1 Ca2+-permeable channel mediates a non-canonical redox adaptation in cancer cells

Abstract Generation of reactive oxygen species (ROS), a natural byproduct of oxygen metabolism, occurs in all aerobic organisms at a controlled rate. Cancer cells are subjected to numerous cellular insults, including dysregulated oncogenes and dissociation from their natural extracellular matrix (ECM) niches, leading to the generation of high levels of ROS. Although cancer cells possess enhanced canonical antioxidant programs that neutralize ROS, they nevertheless exhibit prominent ROS levels in response to these insults, suggesting the existence of additional programs that allow cancer cells to tolerate elevated ROS. In this study, we provide evidence for an unconventional mechanism for redox adaptation involving the ROS-activated, Ca2+-permeable TRPA1 channel, which normally functions as an irritant receptor in sensory neurons but is highly upregulated in breast, lung, malignant neural sheath, and other tumors. We found that TRPA1 overexpression enhances survival and proliferation of MCF-10A cells under conditions of matrix-detachment and this effect was dependent on TRPA1 activation by ROS generated as loss of matrix anchorage. In MCF-10A acini, TRPA1 expression induced Ca2+ entry in ECM-deprived cells in the luminal space and promoted their survival and proliferation. Conversely, TRPA1 knockdown inhibited Ca2+ responses to ROS generated through ECM detachment and induced clearance of cells from the luminal space in breast cancer spheroids. TRPA1 was also activated by ROS-inducing chemotherapies and drove chemoresistance in breast, lung and malignant peripheral nerve sheath tumor cells, and its downregulation suppressed breast xenograft tumor growth and enhanced chemosensitivity. TRPA1 mediated these effects independent of antioxidant responses, but through activation of cellular survival and anti-apoptotic programs involving the RAS-ERK/AKT/mTOR pathways. Together, our findings describe an unexpected mechanism whereby cancer cells co-opt the neuronal TRPA1 channel in order to adapt to oxidative environments. This TRPA1-induced response is distinct from canonical redox adaptation mechanisms that rely on antioxidant program. As TRPA1 inhibitors are currently in clinical trials for pain and respiratory therapies, these studies raise the possibility of using these inhibitors as therapeutic chemosensitizers in TRPA1-enriched tumors. Citation Format: Nobuaki Takahashi, Hsing-Yu Chen, Isaac Harris, Daniel Stover, Roderick Bronson, Thomas Deraedt, Karen Cichowski, Alana Welm, Gordon Mills, Joan Brugge. The TRPA1 Ca2+-permeable channel mediates a non-canonical redox adaptation in cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5478. doi:10.1158/1538-7445.AM2017-5478

Histological and immunohistochemical study of the role of stem cells, conditioned medium and microvesicles in treatment of experimentally induced acute kidney injury in rats

Background: Acute kidney injury (AKI) is a major health problem associated with high morbidity and mortality rates.Mesenchymal stem cells (MSCs) have revealed advantages for therapeutic use in medical practice. Microvesicles (MVs)are membranous, cell-derived vesicles released by MSCs into their microenvironment. The conditioned medium (CM) isthe medium surrounding MSCs. Aim of the Work: This study aimed to investigate the therapeutic potential of MSCs, their CM and microvesicles (MVs) on experimentally induced acute kidney injury in rats. Materials and Methods: Fifty-five rats were divided into five groups: Group I (control group). Group II: given glycerol intramuscularly. Group III: given glycerol then MSCs. Group IV: given glycerol then CM. Group V: given glycerol then MVs. Kidney specimens were processed for H&E and Ki-67 staining and EM studies. Results: Subgroup IIA revealed vacuolation of the cytoplasm, flattening of the epithelial lining the tubules, extrusion of cytoplasm and nuclei into luminal spaces, deeply stained nuclei, and hyaline material in tubular lumina. EM examination of proximal and distal convoluted tubules showed multiple vacuoles, lysosomes, loss of continuity of apical cell membrane,presence of debris in the lumina and vacuolated mitochondria. Group IIB revealed poor improvement with persistence of most lesions. Groups III, VI and V showed amelioration of most of these lesions, and decrease in blood urea and serum creatinine levels. Conclusion: Mesenchymal stem cells, CM and MVs ameliorate induced AKI and with little differences in their effectiveness. CM and MVs can be used in treating diseases.

Stone formation

How stones are retained within the kidney while small in size is still not fully understood. In this talk, we will show two examples of how stones are retained during early growth: One is growth on Randall’s (interstitial) plaque, and the other is growth on mineral that has formed as a luminal plug in a terminal collecting duct. These two mechanisms of stone retention during early growth have distinctive morphologic features that can be seen by methods that show the microscopic structure of the stones. Stones growing on Randall’s plaque display a mineralized region (composed of apatite) that is typically not large in size (less than 0.5 mm across) but which usually shows luminal spaces, which are signs of its origin in the connective tissue of the papilla. Stones growing on ductal plugs also show attachment to a piece of apatite, but the apatite regions are typically larger (often >1 mm long and >0.5 mm wide), and they are solid, without spaces running through them. Still other stone formers exhibit neither of these known mechanisms of stone retention, and we propose urinary stasis as a third possible way that stones are retained within the kidney. We propose that knowing the mechanisms of stone retention during early stone formation should allow for better treatment of stone diseases.

Tu1219 Thin-Poem: Safety and Efficacy of a New Thin Endoscope to Perform Poem Procedure

POEM is a relatively is a safe and effective procedure used to treat patients with Esophageal Achalasia (EA). This technique has demonstrated to be safe, effective and comparable with laparoscopic Heller Myotomy (LHM) at early and mid-term. However during procedure, one of the most difficult steps is to cross below the lower esophageal sphincter (LES) during the submucosal tunnelization, because of the thinness of this space. In fact, “ double endoscope” technique was created in order to check this out during procedure, and this is the site where most of the mucosotomies are presented. Thin endoscopes are used in endoscopy to diagnose and treat diseases where luminal space is compromised with the advantage of a good manipulation. Recently, a nasoendoscope of 5.9mm of outer diameter and 2.4mm of working channel was developed. Out objective was to evaluate the safety and efficacy of this thin endoscope during the POEM procedure.