Angiopoietin-2-induced lymphatic endothelial cell migration drives lymphangiogenesis via the β1 integrin-RhoA-formin axis

Lymphangiogenesis is an essential physiological process but also a determining factor in vascular-related pathological conditions. Angiopoietin-2 (Ang2) plays an important role in lymphatic vascular development and function and its upregulation has been reported in several vascular-related diseases, including cancer. Given the established role of the small GTPase RhoA on cytoskeleton-dependent endothelial functions, we investigated the relationship between RhoA and Ang2-induced cellular activities. This study shows that Ang2-driven human dermal lymphatic endothelial cell migration depends on RhoA. We demonstrate that Ang2-induced migration is independent of the Tie receptors, but dependent on β1 integrin-mediated RhoA activation with knockdown, pharmacological approaches, and protein sequencing experiments. Although the key proteins downstream of RhoA, Rho kinase (ROCK) and myosin light chain, were activated, blockade of ROCK did not abrogate the Ang2-driven migratory effect. However, formins, an alternative target of RhoA, were identified as key players, and especially FHOD1. The Ang2-RhoA relationship was explored in vivo, where lymphatic endothelial RhoA deficiency blocked Ang2-induced lymphangiogenesis, highlighting RhoA as an important target for anti-lymphangiogenic treatments.

     

Cardiovascular and hemodynamic effect of polyethylene glycol in Rats

BACKGROUND: Polyethylene Glycol (PEG) is a solvent and used in a wide range of biomedical applications. Many fatty-acid-based molecules cannot be administered without a solvent in vivo. PEG can be used to dissolve compounds to make them water soluble. However, the effect of PEG on the cardiovascular system has not been studied. In this study, we evaluated the effect of PEG on the cardiovascular system in rat models. METHODS: Twenty male Sprague-Dawley rats weighing 250-300 g were used in this study. The control group (10 rats) were injected intraperitoneally with 0.5 ml of 5% D/W in normal saline and the second group (10 rats) with PEG 400, 2 ml/kg ip, twice a day for 1 week. After 4 weeks, the rats underwent general anesthesia and a 1.4 French ultra miniature pressure volume catheter (Millar catheter) was placed in the left ventricle via the right carotid artery to measure comprehensive hemodynamic data. The data were analyzed with PVAN pressure-volume analysis software. RESULTS: All the systolic and diastolic parameters were similar in both groups except for the effective arterial elastance (Ea), which was decreased in the PEG group. There were no significant differences in maximum (dp/dt(max)) and minimum (dp/dt(min)) development of pressure stroke work, cardiac output, ejection fraction, end systolic volume (Ves), and end diastolic volume. CONCLUSIONS: We have demonstrated that PEG, as a solvent, decreases Ea in rats in comparison to a placebo. Therefore, PEG as a solvent should be used cautiously in the cardiovascular research.     

Respiratory Flow–Sound Relationship During Both Wakefulness and Sleep and Its Variation in Relation to Sleep Apnea

Tracheal respiratory sound analysis is a simple and non-invasive way to study the pathophysiology of the upper airway and has recently been used for acoustic estimation of respiratory flow and sleep apnea diagnosis. However in none of the previous studies was the respiratory flow–sound relationship studied in people with obstructive sleep apnea (OSA), nor during sleep. In this study, we recorded tracheal sound, respiratory flow, and head position from eight non-OSA and 10 OSA individuals during sleep and wakefulness. We compared the flow–sound relationship and variations in model parameters from wakefulness to sleep within and between the two groups. The results show that during both wakefulness and sleep, flow–sound relationship follows a power law but with different parameters. Furthermore, the variations in model parameters may be representative of the OSA pathology. The other objective of this study was to examine the accuracy of respiratory flow estimation algorithms during sleep: we investigated two approaches for calibrating the model parameters using the known data recorded during either wakefulness or sleep. The results show that the acoustical respiratory flow estimation parameters change from wakefulness to sleep. Therefore, if the model is calibrated using wakefulness data, although the estimated respiratory flow follows the relative variations of the real flow, the quantitative flow estimation error would be high during sleep. On the other hand, when the calibration parameters are extracted from tracheal sound and respiratory flow recordings during sleep, the respiratory flow estimation error is less than 10%.     

Serum Levels of IL-6, IL-10, IL-12, IL-17 and IFN-γ and Their Association with Markers of Bone Metabolism in Vitamin D-Deficient Female Students

Different studies have shown the regulatory effects of vitamin D₃ on the immune system and bone metabolism. Regarding the effects of vitamin D on immune cells and the importance of cytokines on bone metabolism, we assessed the association between serum levels of interleukin (IL)-6, IL-10, IL-12, IL-17 and IFN-γ cytokines and bone metabolism markers (Ca, P, PTH, ALP) in female students with vitamin D deficiency compared with control group. A total of 100 subjects with 25-hydroxy vitamin D₃ (25-(OH) D₃) deficiency were selected as case and 100 subjects with sufficient 25-hydroxy vitamin D₃ (25-(OH) D₃) were selected as the control group. The serum levels of IL-6, IL-10, IL-12, IL-17 and IFN-γ were measured by ELISA method. Ionized Ca, PTH, P, ALP levels were also determined in all participants. The results showed a statistically significant positive correlation between the levels of ALP with IFN-γ, PTH with IL-17 and a significant negative correlation between P with IL-10 in vitamin D deficient group. The results suggest that IL-17, IFN-γ and IL-10 are important mediators of bone metabolism and vitamin D affect bone metabolism, at least in part, through immune system. In addition, not only vitamin D affect bone metabolism but also modulates immune responses.     

DCLK1, a promising colorectal cancer stem cell marker,regulates tumor progression and invasion through miR-137 and miR-15a dependent manner

Cancer stem cells (CSCs) are thought to be a major player in tumor initiation, progression, and metastasis. Targeting CSCs for elimination presents a promising therapeutic strategy; however, this approach will require a stronger understanding of CSC biology and identification of CSC-specific markers. The present study was conducted to examine the correlation between DCLK1 and miR-137 and miR-15a levels in colorectal cancer. A total of 222 samples, including 181 colorectal cancer specimens, 24 adenomatosis, and 17 non-adenomatosis colonic polyps, were stained for DCLK1 expression using immunohistochemistry. Also, expression of miR-137 and miR-15a was assessed in colorectal cancer with high and low DCLK1 expression levels. Most colorectal cancer specimens (76%) showed strong expression of DCLK1, whereas only 21% of adenomatous and none of non-adenomatous colonic polyps showed strong DCLK1 expression. A significant difference in DCLK1 expression was found between colorectal cancer, adenomatous, and non-adenomatous colonic polyps (P < 0.001). Higher expression of DCLK1 was more frequently detected in colorectal cases with larger tumor size (P = 0.03), poor differentiation (P = 0.03), and lymph node involvement (P = 0.04). Comparison of miR-137 and miR-15a in colorectal cancer cases revealed a significant inverse correlation with DCLK1 expression (P = 0.03 and P = 0.04, respectively). DCLK1 may act as a candidate marker for colorectal cancer stem cells. The critical role of DCLK1 in colorectal cancer suggests that it may represent an early diagnostic marker and therapeutic target; however, further investigation is warranted.

     

Snoring sounds variability as a signature of obstructive sleep apnea

Snoring sounds vary significantly within and between snorers. In this study, the variation of snoring sounds and its association with obstructive sleep apnea (OSA) are quantified. Snoring sounds of 42 snorers with different degrees of obstructive sleep apnea and 15 non-OSA snorers were analyzed. The sounds were recorded by a microphone placed over the suprasternal notch of trachea, simultaneously with polysomnography (PSG) data over the entire night. We hypothesize that snoring sounds vary significantly within a subject depending on the level of obstruction, and thus the level of airflow. We also hypothesize that this variability is associated with the severity of OSA. For each individual, we extracted snoring sound segments from the respiratory recordings, and divided them into three classes: non-apneic, hypopneic, and post-apneic using their PSG information. Several features were extracted from the snoring sound segments, and compared using a nonparametric statistical test. The results show significant shift in the median of features among the snoring sound classes (p < 0.00001) of an individual. In contrast to hypopneic and post-apneic classes, the characteristics of snoring sounds did not vary significantly over time in non-apneic class. Therefore, we used the total variation norm of each subject to classify the participants as OSA and non-OSA snorers. The results showed 92.9% sensitivity, 100% specificity and 96.4% accuracy.     

Histological and mucin histochemical study of the small intestine of the Persian squirrel (Sciurus anomalus)

This article describes the histological and mucin histochemical properties of the small intestine of the Persian squirrel (Sciurus anomalus). This species is widely distributed in the Middle East and can be found as a companion animal. The histological studies revealed that the plicae circulares were not visible in the tunica mucosa. The maximum height and width of the villi were observed in the duodenum, which then decreased toward the ileum. The muscularis mucosa was scattered, whereas the tunica submucosa was composed of dense connective tissue. The lymphatic nodules were seen in the submucosa of the distal part of the jejunum and ileum, and Brunner’s glands were embedded in the initial portion of the duodenum. The tunica muscularis was significantly thicker in the ileum, and the circular muscle layer was thicker than the longitudinal muscle layer throughout the entire length of the small intestine. The mucin histochemistry, which was examined using the periodic acid-Schiff (PAS) and alcian blue (AB) (pH 1.0 and 2.5) and also PAS–AB (pH 2.5) and aldehyde fuchsin-AB (pH 2.5) techniques coupled with methylation and saponification reaction for some sections, showed that the small intestine mucous content included both carboxylated and sulfated acidic mucins with few neutral mucins. The results of this study contribute to the knowledge of the histological and histochemical characteristics of the gastrointestinal tracts of exotic mammals and provide data for comparison with other mammals.     

The relationship between lymphovascular invasion and angiogenesis,hormone receptors,cell proliferation and survival in patients with primary operable invasive ductal breast cancer

Background

Several well-established tumour prognostic factors are used to guide the clinical management of patients with breast cancer. Lymphovascular invasion and angiogenesis have also been reported to have some promise as prognostic factors. The aim of the present study was to examine the prognostic value of tumour lymphovascular invasion and microvessel density compared with that of established prognostic factors in invasive ductal breast cancer.

Methods

In addition to hormone receptor status and Ki-67 proliferative activity, lymphovascular invasion and microvessel density and their relationship with survival were examined in patients with invasive ductal breast cancer. Full sections and tissue microarrays (n?=?384 patients) were utilised to assess these factors and were scored by appropriate methods.

Results

On univariate analysis tumour size (P?<?0.05), lymph node involvement (P?<?0.01), lymphovascular invasion (P?<?0.05), microvessel density (P?<?0.05) and local- regional treatment (P?<?0.01) were associated with poorer survival in ER negative tumours. On multivariate analysis in ER negative tumours lymph node involvement (P?<?0.01) and local- regional treatment (P?<?0.05) were independently associated with poorer cancer-specific survival. On univariate analysis tumour grade (P?<?0.05), lymph node involvement (P?<?0.001), HER-2 (P?<?0.05), Ki-67 (P?<?0.01) and lymphovascular invasion (P?<?0.001) were associated with poorer survival in ER positive tumours. On multivariate analysis lymph node involvement (P?<?0.001), Ki-67 (P?<?0.001) and lymphovascular invasion (P?<?0.05) were independently associated with poorer cancer-specific survival in ER positive tumours.

Conclusion

Lymphovascular invasion but not microvessel density was independently associated with poorer survival in patients with ER positive but not ER negative invasive ductal breast cancer.

     

Elevated IL-38 Serum Levels in Newly Diagnosed Multiple Sclerosis and Systemic Sclerosis Patients

ObjectiveInterleukin (IL)-38 is a newly discovered member of the IL-1 cytokine family with a proposed anti-inflammatory profile. We studied the probable role of this cytokine in the pathogenesis of two autoimmune diseases: multiple sclerosis (MS) and systemic sclerosis (SSc).Subjects and MethodsA total of 87 MS patients and 86 SSc patients (40 new and recently untreated cases and 46 treated cases) were selected for this study. Eighty-seven and 80 age- and sex-matched healthy subjects were included as controls for MS and SSc, respectively. Clinical and paraclinical features of the patients were recorded at the time of sampling. Serum IL-38 was measured by ELISA.ResultsLevels of serum IL-38 did not significantly differ between the total MS or SSc patients compared to controls. However, levels of IL-38 were significantly higher in newly diagnosed patients of MS (206.43 ± 38.97 pg/mL, p < 0.0001) than in those previously treated (158.04 ± 39.45 pg/mL). Similarly, new/recently untreated cases of SSc patients showed increased IL-38 levels (185.19 ± 36.27 pg/mL, p = 0.001) compared to treated patients (166.82 ± 33.08 pg/mL). IL-38 levels in newly diagnosed MS patients (p = 0.007) and new/recently untreated SSc patients (p = 0.032) were significantly higher than those in healthy controls.ConclusionThe higher serum levels of IL-38 in new or recently untreated cases of MS and SSc patients than in treated patients and healthy controls suggest the possible role of this cytokine in the development of these diseases or as part of a feedback loop to attenuate the inflammatory conditions in early stages of these diseases.     

Differentiation of Wharton's jelly mesenchymal stem cells into neurons in alginate scaffold

Alginate scaffold has been considered as an appropriate biomaterial for promoting the differentiation of embryonic stem cells toward neuronal cell lineage. We hypothesized that alginate scaffold is suitable for culturing Wharton's jelly mesenchymal stem cells(WJMSCs) and can promote the differentiation of WJMSCs into neuron-like cells. In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 m M Ca Cl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/m L basic fibroblast growth factor. These cells were also cultured in conventional two-dimensional culture condition in the presence of neurogenic medium as controls. After 10 days, immunofluorescence staining was performed for detecting β-tubulin(marker for WJMSCs-differentiated neuron) and CD271(motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition. These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.