Backgrounds and aim
Anastomotic leak (AL) following esophagectomy for esophageal cancer (EC) remains an important cause of prolonged hospitalization and impaired quality of life. Recently, ...indocyanine green (ICG) fluorescein imaging has been used to evaluate the gastric conduit blood supply during EC surgery. Although several factors have been reported to be associated with AL, no studies have evaluated the relationships between risk factors for AL, including ICG fluorescein imaging. The purpose of this study was to investigate the risk factors associated with AL following esophagectomy and to evaluate the impact of ICG fluorescein imaging of the gastric conduit during EC surgery.
Methods
One hundred and twenty patients undergoing esophagectomy with esophagogastric anastomosis for EC were enrolled in this retrospective study. Clinicopathological factors, preoperative laboratory variables, and surgical factors, including ICG fluorescence imaging, were analyzed to determine their association with AL. Univariate and multivariate logistic regression analysis was used to evaluate the impact of each of these factors on the incidence of AL.
Results
Among the 120 patients enrolled in the study, 10 (8.3%) developed AL. Univariate analysis demonstrated an increased risk of AL in patients with a high-neutrophil-to-lymphocyte ratio (
p
= 0.0500) and in patients who did not undergo ICG fluorescein imaging (
p
= 0.0057). Multivariate analysis revealed that the absence of ICG imaging was an independent risk factor for AL (
p
= 0.0098).
Conclusions
Using ICG fluorescein imaging to evaluate blood flow in the gastric conduit might decrease the incidence of AL following EC surgery.
We developed a sensitive quantitative assay for detecting Ralstonia solanacearum in soil by most probable number (MPN) analysis based on bio-PCR results. For development of the detection method, we ...optimized an elution buffer containing 5 g/L skim milk for extracting bacteria from soil and reducing contamination of polymerase inhibitors in soil extracts. Because R. solanacearum can grow in water without any added nutrients, we used a cultivation buffer in the culture step of the bio-PCR that contained only the buffer and antibiotics to suppress the growth of other soil microorganisms. To quantify the bacterial population in soil, the elution buffer was added to 10 g soil on a dry weight basis so that the combined weight of buffer, soil, and soil-water was 50 g; 5 mL of soil extract was assumed to originate from 1 g of soil. The soil extract was divided into triplicate aliquots each of 5 mL and 500, 50, and 5 μL. Each aliquot was diluted with the cultivation buffer and incubated at 35 °C for about 24 h. After incubation, 5 μL of culture was directly used for nested PCR. The number of aliquots showing positive results was collectively checked against the MPN table. The method could quantify bacterial populations in soil down to 3 cfu/10 g dried soil and was successfully applied to several types of soil. We applied the method for the quantitative detection of R. solanacearum in horticultural soils, which could quantitatively detect small populations (9.3 cfu/g), but the semiselective media were not able to detect the bacteria.
The biochemical transduction of excitatory synaptic signals occurs in the cytoplasm within dendritic spines. The associated reaction kinetics are shaped by the mobility of the signaling molecules; ...however, accurate monitoring of diffusional events within the femtoliter-sized spine structures has not yet been demonstrated. Here, we applied two-photon fluorescence correlation spectroscopy and raster image correlation spectroscopy to monitor protein dynamics within spines, revealing that F-actin restricts the mobility of proteins with a molecular mass of >100 kDa. This restriction is transiently removed during actin remodeling at the initial phase of spine structural plasticity. Photobleaching experiments combined with super-resolution imaging indicate that this increase in mobility facilitates molecular interactions, which may modulate the functions of key postsynaptic signaling molecules, such as Tiam1 and CaMKII. Thus, actin polymers in dendritic spines act as precise temporal regulators of molecular diffusion and modulate signal transduction during synaptic plasticity.
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•Two-photon FCS and RICS report protein dynamics within dendritic spines•Actin polymers within spines restrict molecular mobility in a size-specific manner•Actin remodeling during the initial phase of structural LTP increases molecular mobility•Actin polymers in spines act as precise temporal regulators of postsynaptic signaling
Obashi et al. show that actin polymers within dendritic spines restrict mobility of large molecules using optical measurements of fluorescence correlation. Acute actin remodeling induced by plasticity-inducing stimuli increases the mobility of large postsynaptic signaling molecules, which regulate long-term changes in synaptic property.
Although there is a growing interest in developing circulating microRNA (miRNA) as noninvasive diagnostic biomarkers for the detection of high-risk colorectal adenomas and early-stage CRCs, but the ...comparative diagnostic significance of serum vs. exosomal miRNAs remains unexplored.
Based upon published literature, we performed an initial discovery step by investigating the expression of a miRNA panel in 20 normal colonic mucosa, 27 adenomas, and 19 CRC tissues. We performed subsequent validation by quantifying expression of candidate miRNAs in total serum and in exosomes from 26 adenoma patients and 47 healthy controls, and evaluated their clinical significance and potential diagnostic value in colorectal adenomas.
We observed that the expression of four miRNAs, miR-21, miR-29a, miR-92a, and miR-135b, was significantly higher in colorectal adenomas vs. normal colonic mucosa. During validation, expression of miR-21, miR-29a and miR-92a in serum was significantly higher in adenomas vs. healthy controls, significantly correlated with adenoma size and total adenoma number within the colorectum, and significantly discriminated patients with advanced adenomas. In contrast, although exosomal miR-21 and miR-29a levels in adenoma patients were significantly higher than those of healthy volunteers, only exosomal miR-21 significantly correlated with adenoma size and total adenoma number, and could discriminate patients with high-risk adenomas.
Compared to exosomal miRNAs, serum levels of miR-21, miR-29a and miR-92a are superior diagnostic biomarkers in patients with high-risk adenomatous polyps.
The primordium of the exoskeleton of an insect is epithelial tissue with characteristic patterns of folds. As the insect develops from larva to pupa, the spreading of these folds produces the ...three-dimensional shape of the exoskeleton of the insect. It is known that the three-dimensional exoskeleton shape has already been encoded in characteristic patterns of folds in the primordium; however, a description of how the epithelial tissue forms with the characteristic patterns of folds remains elusive. The present paper suggests a possible mechanism for the formation of the folding pattern. During the primordium development, because of the epithelial tissue is surrounded by other tissues, cell proliferation proceeds within a confined geometry. To elucidate the mechanics of the folding of the epithelial tissue in the confined geometry, we employ a three-dimensional vertex model that expresses tissue deformations based on cell mechanical behaviors and apply the model to examine the effects of cell divisions and the confined geometry on epithelial folding. Our simulation results suggest that the orientation of the axis of cell division is sufficient to cause different folding patterns in silico and that the restraint of out-of-plane deformation due to the confined geometry determines the interspacing of the folds.
Abstract
Epithelial folding is a universal biological phenomenon in morphogenesis, typical examples being brain gyri, villi of the intestinal tract, and imaginal discs in invertebrates. During ...epithelial morphogenesis, the physical constraints imposed by the surrounding microenvironment on epithelial tissue play critical roles in folding morphology. In this study, we focused on the asymmetry of the environmental constraints sandwiching the epithelial sheet and introduced the degree of asymmetry, which indicates whether the basal or apical side of the epithelium is closer to the constraint wall. Then, we investigated the relationship between the degree of asymmetry and epithelial folding morphology using three-dimensional vertex simulations. The results show that the folding patterns of the epithelial sheets change from spot patterns to labyrinth patterns and then to hole patterns as the degree of asymmetry changes. Furthermore, we examined the pattern formation in terms of the equation of out-of-plane displacement of the sheet derived from the mechanical energy functional.
In biological development, multiple cells cooperate to form tissue morphologies based on their mechanical interactions; namely active force generation and passive viscoelastic response. In ...particular, the dynamic processes of tissue deformations are governed by the viscous properties of the tissues. These properties are spatially inhomogeneous because they depend on the tissue constituents, such as cytoplasm, cytoskeleton, basement membrane and extracellular matrix. The multicellular mechanics of tissue morphogenesis have been investigated in vertex dynamics models. However, conventional models are applicable only to quasi-static deformation processes, which do not account for tissue viscosities. We propose a vertex dynamics model that simulates the viscosity-dependent dynamic deformation processes during tissue morphogenesis. By incorporating local velocity fields into the governing equation of vertex movements, the model turns Galilean invariant. In addition, the viscous properties of tissue components are newly expressed by formulating friction forces on vertices as functions of the relative velocities among the vertices. The advantages of the proposed model are examined by epithelial growth simulations under the employed condition for quasi-static processes. As a result, the epithelial vesicle simulated by the proposed model is linearly elongated with nearly free stress, while that simulated by the conventional model is undulated with compressive residual stress. Therefore, the proposed model is able to reflect the timescale of deformations by satisfying Galilean invariance. Next, the applicability of the proposed model is assessed in epithelial growth simulations of viscous extracellular materials. In this test, the epithelial vesicles are deformed into tubular shapes by oriented cell divisions, and their morphologies are extremely sensitive to extracellular viscosity. Therefore, the dynamic deformations in the proposed model depend on the viscous properties of tissue components. The proposed model will be useful for simulating dynamic deformation processes of tissue morphogenesis depending on viscous properties of various tissue components.
Three semi-selective media, DTarTA, SPbc, and SPamt, were developed and tested to isolate
Pseudomonas syringae
pv.
maculicola
(
Psm
) and
P
.
cannabina
pv.
alisalensis
(
Pca
) from
Raphanus sativus
...seeds. DTarTA contained D-tartaric acid as a carbon source and potassium tellurite, ampicillin sodium, and methyl violet as antibiotics. DTarTA suppressed growth in 19 of the 24 pathovars from the
P
.
syringae
complex, whereas
Psm
and
Pca
grew and formed gray to black colonies. SPamt contained sucrose and peptone as nutrient sources and was supplemented with bromothymol blue and the same antibiotics present in DTarTA and
Psm
and
Pca
formed yellowish to dark brown colonies on the SPamt medium. SPbc contained sucrose and peptone and was supplemented with cephalexin and boric acid as antibiotics and
Psm
and
Pca
formed semi-translucent to white colonies on the SPbc medium. SPamt and SPbc suppressed the growth of several plant-associated bacteria (except the
P
.
syringae
complex). The growth of saprophytic bacteria in seeds on the different media was compared with that on King’s B medium, using five types of commercially available
Raphanus sativus
seeds. The suppression rate of DTarTA was 85–99% and was lower for seeds with more saprophytic bacteria. The suppression rates of SPamt and SPbc were 90–99%. In detection tests using 10,000 seed samples mixed with
Pca
or
Psm
-contaminated seeds, it was possible to selectively isolate
Psm
and
Pca
using SPamt and SPbc, even when the colony numbers of the target bacterium constituted less than 10% of the total colonies.
Key points
•
Bacterial leaf spot and blight pathogens were selectively isolated from seeds
.
•
DTarTA medium distinguishes these pathogens from P. syringae complex pathovars
.
•
SPamp and SPbc media have different selectivity for plant-associated bacteria
.
Background
Rac GTPase activating protein 1 (RacGAP1) plays a regulatory role in cell growth, transformation and metastasis. The aim of this study was to clarify the association between RacGAP1 ...expression and clinical outcome in patients with gastric cancer.
Methods
A total of 232 gastric cancer patients in our institute who underwent surgery without preoperative treatments were enrolled in this study. We investigated RacGAP1 expression using immunohistochemistry (IHC) and evaluated IHC scores calculated by the percentage of positive cells and intensity and its expression at the invasive front.
RACGAP1
expression was also assessed.
Results
RacGAP1 expression was observed in the nuclei of gastric cancer cells. Evaluation by IHC score showed no significant correlations with clinicopathological variables except for histological differentiation. In transcriptional analyses,
RACGAP1
expression was elevated in diffuse type gastric cancer than intestinal type without a significant difference. We observed significant correlations of RacGAP1 protein expression at the invasive front with older age, tumor size, lymph node metastasis, lymphatic invasion, vascular invasion and advanced stage. Patients with RacGAP1 protein expression at the invasive front had significantly poorer prognosis than those without it (
P
< 0.0001). In multivariate analysis, lymph node metastasis, distant metastasis and positive RacGAP1 expression at the invasive front were independent prognostic factors (lymph node metastasis:
P
= 0.0106; distant metastasis:
P
= 0.0012; RacGAP1:
P
= 0.0011).
Conclusions
RacGAP1 expression at the invasive front in gastric cancer was significantly correlated with factors reflecting tumor progression and poor prognosis. Our data suggest that RacGAP1 might play important roles in the progression of gastric cancer.
Purpose
The coexistence of sarcopenia is associated with postoperative complications, including infection after abdominal surgery. We evaluated the association between sarcopenia and surgical site ...infection (SSI) after surgery for ulcerative colitis.
Methods
The subjects of this retrospective study were 69 patients who underwent restorative proctocolectomy with perioperative abdominal computed tomography (CT). Sarcopenia was diagnosed by measuring the cross-sectional area of the right and left psoas muscles as the total psoas muscle area on CT images. We assessed whether sarcopenia was associated with SSI and clinical factors, including nutritional and inflammatory markers.
Results
The lowest quartiles defined as sarcopenia in men and women were 567.4 and 355.8 mm
2
/m
2
, respectively. According to this classification, 12 men and 6 women had sarcopenia. Patients with sarcopenia had a lower body mass index (
p
= 0.0004) and a higher C-reactive protein concentration (
p
= 0.05) than those without sarcopenia. SSIs were identified in 12 patients (17.3 %) and included six pelvic abscesses and seven wound infections. According to multivariate analysis, sarcopenia was an independent risk factor for SSI (odds ratio = 4.91, 95 % confidence interval 1.09–23.5,
p
= 0.03).
Conclusion
Sarcopenia is predictive of SSI after pouch surgery for ulcerative colitis.
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