bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Morphological Features of Fibrillar Collagen Indicative of Tumor Progression in Bladder Cancer
Authors: Borhani Soheila , Macias Virgilia , Kajdacsy‐Balla Andre 
Published: May 2022

ObjectiveBladder cancer is the most common malignancy involving the urinary system. As with all types of cancer, accurate monitoring of stage progression is critical in determining proper treatment strategies and predicting clinical outcomes. Collagen fibers are a major component of tumor microenvironment, and their post‐translational modifications during the course of tumor progression are well‐documented in a number of malignancies including bladder cancer. In the present feasibility study we sought to determine whether quantitative morphological features extracted from collagen fibers can be used to distinguish between urothelial carcinoma tumors pre‐muscle‐invasion (stage pT1) and post‐muscle‐invasion (stage pT2).MethodsPicroSirius Red (PSR) staining was used to detect fibrillar collagen in 8 tissue microarray (TMA) cores acquired from Biomax (4 stage pT1, 4 stage pT2). We applied auto‐fluorescence imaging techniques to detect the PSR signal, and employed the CT‐Fire freeware to automatically extract individual collagen fibers. Five morphological features (including length, width, straightness, orientation, and anisotropy) were computed for each detected fiber. Individual features were then combined within each slide to form five corresponding histograms per TMA core. Finally, for each histogram we computed two measures of central tendency (i.e., mean and median), two measures of spread (i.e., range and standard deviation), and two measures of symmetry (i.e., skewness and kurtosis).ResultsTo identify any statistically‐significant differences between the pT1 and pT2 groups, we employed the student’s t‐test (run in Python 3.8.5) to compare the two groups with respect to each of the five morphological features and across each of the six distributional measures mentioned in the Methods section (30 comparisons in total). Of these 30 comparisons, all but one (i.e., median) of the six straightness measures resulted in a p‐value < 0.05. Further, each of the two measures of spread (i.e., range and standard deviation) of straightness returned a p‐value < 0.001. We are currently working on expanding the size of our dataset to further validate these results.ConclusionOur preliminary findings are in line with the hypothesis that collagen remodeling is strongly associated with muscle‐invasive bladder cancer, and suggest that quantitative features measuring the straightness of individual fibers may be used to improve tumor staging. This type of quantitative approach is particularly helpful in distinguishing between stage pT1 and stage pT2 of bladder cancer, a task which can be challenging at times even for experienced pathologists.
The ion binding site in transmembrane domain 5 of NBCe1 has negligible role in electrogenicity
Authors: Choi Inyeong , Lee Soojung 
Published: May 2022

BackgroundThe sodium bicarbonate transporters move HCO3– across cell membranes and regulate intracellular pH or transepithelial HCO3– transport. Some members of these transporters, such as NBCe1 and NBCe2, mediate electrogenic transport (1 Na+ and 1 CO32– or 2 HCO3–), while others, such as NBCn1 and NBCn2, mediate electroneutral transport (1 Na and 1 HCO3–). A structure/function study has shown that the loop between transmembrane (TM) 7 and 8 is responsible for electrogenicity. On the other hand, a recent report on the molecular dynamics simulations of NBCe1 cryoEM structure has proposed an ion binding site, which is located in transmembrane domain (TM) 5 and serves as a transient binding site to attract anions from the surrounding solution. In the present study, we tested whether this binding site is responsible for electrogenic Na/HCO3 transport.MethodsAmino acid residues constituting the ion binding site of NBCe1 and NBCn1 were mutated and their effects on electrogenicity were evaluated by expressing the corresponding mutant transporters in Xenopus oocytes and assessing an Na/HCO3 transport current (INBC) in two‐electrode voltage clamp. In addition, the loop between TM7 and TM8 was replaced and its effect on INBC was examined.Results1) Replacing TM5 of NBCe1 with the corresponding TM5 of NBCn1 retained INBC, whereas a reciprocal replacement did not. 2) Replacing charged residues constituting the ion binding site in NBCn1 with the residues at the corresponding site in NBCe1 did not induce INBC. 3) Replacing the loop between TM7 and TM8 in NBCe1 with the corresponding loop in NBCn1 abolished INBC. 4) Replacing the loop in the NBCe1/TM5 chimeric transporter with the loop in NBCn1 abolished INBC.ConclusionWe conclude that the NBCe1 electrogenicity is not associated with the ion binding site but determined by the loop between TM7 and TM8.
A Knuckle‐Walking Signal in the Trabecular Architecture of the African Ape Scaphoid
Authors: Goldstein Deanna M. , Patel Biren A. , Sylvester Adam D. 
Published: May 2022

African ape (Panand Gorilla) carpals have few diagnostic features in their external morphology reflecting knuckle‐walking hand postures, and whether the internal trabecular architecture of their carpals yields a functional signal for knuckle‐walking is unclear. Here we perform a high‐density, multiple‐VOI analysis of trabecular architecture in the scaphoid of knuckle‐walking African apes and non‐knuckle‐walkers (Pongo, Macaca,and Papio). We predict that (1) Gorilla, the taxon with largest body mass, will have the highest trabecular thickness (Tb.Th) and trabecular spacing (Tb.Sp) because Tb.Th and Tb.Sp have been shown to be correlated with body size, (2) bone volume fraction (BV/TV) will be higher in habitually terrestrial taxa than in arboreal taxa, as the forelimb is thought to experience relatively high compressive loads during terrestrial locomotion, (3) Pongo will have the lowest degree of anisotropy (DA) as they engage in variable locomotor behaviors, and (4) pattern of DA distribution will differ between knuckle‐walking and non‐knuckle‐walking taxa, as the consistency and direction of wrist joint loading likely differs among primates with different quadrupedal hand postures.Fifty‐four scaphoids were scanned using high‐resolution µCT (voxel range: 48‐27 μm). Surface models were created from image data and then subjected to a weighted spherical harmonic analysis. Volumes of interest (VOIs) were manually placed into a surface model of the average scaphoid, and then warped to individual specimens using the thin plate spline interpolation function. Trabecular properties were calculated using the BoneJ plugin for ImageJ and MATLAB, and variation in distribution of trabecular parameters among taxa was summarized using a principal component analysis (PCA). Parameter values among species were compared with non‐parametric MANOVAs using a randomized residual permutation procedure, and PC scores were compared using Kruskal‐Wallis ANOVAs. The parameter distribution within each bone was visualized by comparing species‐average color maps of average parameter values.As predicted, Tb.Th and Tb.Sp are greatest in Gorilla, indicating that these parameters are related in part to body size. In contrast, results indicate that BV/TV and DA magnitudes do not follow predictions. Specifically, habitually terrestrial taxa do not have significantly higher BV/TV than arboreal taxa, and taxa that engage in more variable locomotor behaviors (i.e., Pongo) do not have the lowest values of DA. Finally, results indicate that knuckle‐walking African apes can be distinguished from other taxa studied here by pattern of DA distribution, and these differences appear to reflect differences in habitual hand postures and locomotor modes. If this pattern is unique to knuckle‐walking apes among hominoids, and is also identified in the scaphoid of fossil hominoids, then it may be used to indicate whether fossil taxa were habitual knuckle‐walkers.
Telocytes: Newly Discovered Interstitial Cells in the Human Female Internal Reproductive Organs – Identification of a Suitable Immunohistochemical Marker and Possible Significance in the Pathogenesis of Uterine Fibroids
Authors: Klein Martin , Csöbönyeiová Mária , Visnyaiová Kristína , Danišovič Ľuboš , Varga Ivan 
Published: May 2022

IntroductionTelocytes are a cell population discovered in 2005. Within the organs of the female reproductive system (FRS), telocytes were described as having an immunomodulatory role and also functioning as hormone sensors. From the pathological perspective, telocytes were described in the pathogenesis of a wide spectrum of different conditions, including a role in tumorigenesis. The gold standard of telocyte research is electron microscopy, but immunohistochemistry is also widely applied. The main objective was to identify the most suitable immunohistochemical marker and subsequently use it in the study of uterine specimens from patients with uterine fibroids (leiomyomas).Material andMethodsUterine specimens were obtained from 26 patients who underwent hysterectomy for various indications, including ectopic pregnancy and uterine fibroids. From all the uteruses, representative specimens were obtained from the myometrium and endometrium of the uterine body, and also from the endo‐ and exocervical mucosa. 5 μm thick sections were further processed for immunohistochemistry. To identify the most suitable marker of telocytes, 8 primary antibodies were used: CD117/c‐kit, CD34, vimentin, α‐smooth muscle actin, progesterone and estrogen receptor, desmin and S100 protein (DAKO, Denmark).ResultsThe most specific marker was c‐kit. C‐kit‐positive cells were spindle‐shaped, triangular and star‐shaped. C‐kit positive cells were located in the endometrium, myometrium, exocervical, and endocervical mucosa. Comparing the distribution within the individual parts of the uterus, the most numerous population was observed within the myometrium. A considerable number of telocytes was also observed within the endocervical mucosa, while the fewest telocytes were described in the endometrium and exocervical mucosa. In the specimens of uterine fibroids, the non‐affected parts of the myometrium contained c‐kit positive cells within all the myometrial sublayers. The average c‐kit positivity expressed in percentage was 2.19 ± 0.51%. Those parts of the myometrium affected by uterine fibroids displayed complete c‐kit negativity.ConclusionC‐kit is the most suitable marker in the research of telocytes in the FRS. The loss of telocytes in the myometrium may have a role in the pathogenesis of uterine fibroids via three mechanisms: 1) hormonal imbalance, 2) stem cell dysregulation and 3) diminished angiogenesis. Therefore, therapeutic targeting of telocytes can be a potential future approach in the treatment of this condition.
In utero exposure to dietary emulsifier alters the behavioral activity in offspring
Authors: Mishra Jayshree 
Published: May 2022

ObjectivePediatric mental health disorder is a combination of neuroinflammatory conditions with unknown etiology and currently, there are less awareness on the impacts of food additives on pediatric neuroinflammatory conditions and adolescent mental health disorders. Food additives such as dietary emulsifier (DE) (polysorbate 80, carrageenan and maltodextrin) are added to a variety of processed foods, and their consumption has increased dramatically worldwide which could be a risk factor for pediatric mental health disorders. Therefore, there is a critical need to understand the impact of maternal exposure to food additives associated pediatric mental health and the underlying mechanism.Materials and MethodsBehavioral testing such as tail flick assay, radial arm, rotarod and hot plate assay was done on the offspring from both control and DE exposed mice for the impaired motor activity, pain sensitivity and cognitive behavior. Colonic microbial composition was determined through 16sRNA pyrosequencing of fecal DNA samples from control and in utero DE‐exposed offspring. Level of pro inflammatory cytokines in the colon tissue as well as the serum sample was measured using Multianalyte cytokine assay kit from Qiagen.Summary of ResultsOur results show that in utero exposure to DE leads to decreased brain sizes and impaired motor learning in the offspring. Additionally, in utero exposure to DE leads to decreased pain sensitivity as a sign of stress in offspring. In utero exposure to DE shows on onset of cognitive impairment in offspring. In utero exposure to DE leads to an elevated M1 macrophage cytokine profile and CLGI symptoms in the gastrointestinal tract of offspring. In utero exposure to DE leads to gut dysbiosis in offspring. Among the DE‐exposed adolescents, the male offspring had significantly higher impact on these parameters compared to the DE‐exposed female offspring. In utero exposure to DE leads to imbalance in serum levels of dihydrotestosterone (DHT) and estradiol (E2) in the Offspring.ConclusionOur data suggest that maternal exposure to food additive is associated with the male biased cognitive impairment and neurobehavioral changes in the offspring. In future, we will determine the specific mechanism responsible for the sex specific alternation in offspring born to DE exposed mother to develop a preventative and therapeutic strategies.
Limb Hemodynamic Responses to a Single Bout of Inspiratory Muscle Strength Training
Authors: Plouffe Audrey A. , Vranish Jennifer R. 
Published: May 2022

BackgroundInspiratory muscle strength training (IMST) has been shown to bring about improvements in respiratory muscle strength and reductions in blood pressure after 6 weeks of daily high intensity training. However, there has been little investigation into the effects of IMST on limb blood flow, and it is unclear how hemodynamics are altered within a single bout of training.PurposeTo investigate the acute effects of one session of either low or high intensity IMST on blood flow in the upper and lower limbs.MethodsParticipants (n=7, 57% female) aged 20 to 47 years completed two randomized laboratory visits wherein resting blood flow was measured via duplex Doppler ultrasound at the left brachial and popliteal arteries. Following baseline measures, participants' maximal inspiratory pressure (PImax) was measured using a PowerBreathe K3 device. Participants performed one session (30 breaths) of IMST at either 15% or 75% of their PImax, and blood flow was measured again at each artery for five breaths during the final ten breaths of the training. At their second visit, subjects performed whichever training level had yet to be completed and blood flow was assessed as described above.ResultsMean blood flow at the brachial artery appeared reduced during both 15% (p=0.070) and 75% training (p=0.078), which may be attributed to a significant increase in retrograde blood flow in both cases (p=0.014 and p=0.037, respectively). This increase in retrograde blood flow tended to be greater during the 75% training (p=0.066). Popliteal artery blood flow showed similar trends to brachial artery blood flow.ConclusionsWhile IMST appears to reduce brachial artery mean blood flow during a single training session, this may be due to alterations in retrograde blood flow, which tended to increase in an intensity‐dependent manner. Further investigation into the chronic hemodynamic effects of IMST is warranted.
Role of inflammation and ER stress in HFpEF pathogenesis
Authors: Srinivas Balaji , O'Regan Jacob D. , Belmadani Souad , Matrougui Khalid 
Published: May 2022

BackgroundHeart failure with preserved ejection fraction (HFpEF), characterized by diastolic dysfunction, inflammation, cardiac hypertrophy and fibrosis, myocardial capillary rarefaction, pulmonary hypertension, aortic stiffness, and impaired endothelial function, is the major unmet need in cardiovascular medicine and remains an untreatable cardiovascular disease. The role of inflammation and endoplasmic reticulum (ER) stress in HFpEF pathogenesis is not well understood.MethodsC57/Bl6 mice were divided into five groups. Group 1: control mice. Group 2: Mice fed high fat diet (HFD) and received L‐NAME (eNOS inhibitor,0.5 g/L) in the drinking water for 4 months. Group 3: Mice fed HFD + L‐NAME for 4 months then treated with ER stress inhibitor for one month (Tudca, 150 mg/Kg). Group 4: Mice fed HFD + L‐NAME then treated with anakinra for one month (IL‐1beta receptor antagonist, 1 µg/mouse/every other day). Group 5: Mice fed HFD + L‐NAME then treated with Tudca + anakinra for one month. We measured body weight, blood pressure, cardiac function, diabetes status, and vascular endothelial function.ResultsMice in group 2 fed HFD and L‐NAME in the drinking water for 4 months developed metabolic syndrome, hypertension, cardiac hypertrophy, lung edema, low running performance, vascular endothelial dysfunction, cardiac diastolic dysfunction, but no change in cardiac ejection fraction (EF) compared to group 1. Interestingly, the treatment of mice in groups 3, 4, and 5, with Tudca with and without anakinra provides protection from hypertension, body weight increase, metabolic syndrome, cardiac hypertrophy, lung edema, and vascular endothelial and diastolic dysfunction induced by HFD and L‐NAME.ConclusionOur data illustrate that ER stress and inflammation play a critical role in HFpEF pathogenesis.
Inhibition of Ileal Bile Acid Transport is Protective Against Cholestatic Liver Injury in Cyp2c70‐/‐Mice with Humanized Bile Acid Composition
Authors: Truong Jennifer K. , Donepudi Ajay , Bennett Ashley , Malla Sudarshan , Zaufel Alex , Pachura Kimberly , Hellen Dominick , Moustafa Tarek , Dawson Paul , Karpen Saul 
Published: May 2022

Cyp2c70 is the key murine liver enzyme responsible for the synthesis of 6‐hydroxylated muricholates. Cyp2c70 knockout (2c70‐/‐) mice present with a more human‐like bile acid (BA) composition devoid of protective, hydrophilic muricholic acids and with features of cholestatic liver injury. Pharmacological inhibition of the ileal BA transporter (IBAT) has been shown to be protective in cholestatic models such as the Mdr2‐/‐ mouse. The aims of this study were to determine if IBAT inhibition is protective in 2c70‐/‐ mice and identify the underlying molecular mechanisms.Methods2c70‐/‐ mice (C57Bl/6J) were generated using CRISPR/Cas9 technology. Male and female wild type (WT) and 2c70‐/‐ mice were fed chow or chow containing 0.006% SC‐435 for 9 weeks following weaning at 3 weeks of age. Standard growth parameters were recorded; organs, gallbladder bile and blood were collected at necropsy for histologic, RNA and biochemical analyses.ResultsMale and female 2c70‐/‐ mice maintained on chow diet exhibited increased levels of serum liver injury markers ALT, AST, and BAs compared to WT mice. Evaluation of 2c70‐/‐ liver histology revealed increased hepatic inflammation, macrophage infiltration and bilary cell proliferation, which were markedly reduced with SC‐435 treatment. IBAT inhibition in 2c70‐/‐ mice improved liver histology, and reduced hepatic inflammation and bilary cell proliferation to levels indistinguishable from WT. The liver BA content was increased by 53% in 2c70‐/‐ mice, and was enriched in more hydrophobic BA species with an overall hydrophobicity similar that of humans (Figure 1). Treating 2c70‐/‐ mice with SC‐435 reduced the hepatic BA burden by 61%, but did not affect the overall hydrophobicity. RNA‐Seq and KEGG pathway analysis performed using liver RNA revealed genes involved in hepatic steroid and lipid metabolism (Sult2a1, Sult1e1, Hsd17b7), and detoxification (Cyp2c39, Cyp2c54, Cyp2c59, Cyp3a41a/b) were down‐regulated in 2c70‐/‐ mice and their expression were rescued with SC‐435 treatment. Conversely, genes in pathways related to immune cell activation and inflammation were significantly up‐regulated in 2c70‐/‐ mice and reduced to WT levels with IBAT inhibition (Figure 2).ConclusionsOur findings indicate that reducing the hepatic BA burden without altering the hydrophobic compostion is sufficient to protect 2c70‐/‐ mice from hepatic injury. The findings suggest that there may be a threshold in liver for pathological accretion of hydrophobic BAs and reducing the hepatic BA pool is sufficient to alleviate liver injury, independent of hydrophobicity. Moreover, these data support the inhibition IBAT activity, and other approaches to reduce hepatic return of BAs in the enterohepatic circulation, as therapeutic targets to treat cholestasis.
Sex‐dependent Regulation of Mitochondrial Respiratory Function in Mouse Brain Microvessels by Peroxynitrite Decomposition Catalyst
Authors: Sakamuri Siva S. , Sure Venkata N. , Kolli Lahari , Wisen William P. , Evans Wesley R. , Lindsey Sarah H. , Mostany Ricardo , Katakam Prasad V. 
Published: May 2022

BackgroundPeroxynitrite (PN) is a strong oxidizing and nitrating molecule. PN is a potent inhibitor of mitochondrial respiration and promotes ischemia‐reperfusion injury following stroke. In isolated mouse brain mitochondria, we observed that PN donors inhibit mitochondrial respiratiory function whereas the PN decomposition catalyst, Fe (III) tetrakis (1‐methyl‐4‐pyridyl) porphyrin pentachlorideporphyrin pentachloride (FeTMPyP), enhances mitochondrial state III and state IVo mitochondrial respiration. In addition, we demonstrated that mitochondrial respiration is the primary contributor of cellular energy in brain microvessels (BMVs). The present study tested the hypothesis that FeTMPyP negatively regulates mitochondrial respiration in the mouse BMVs.MethodsBMVs were isolated from male and female mice (C57Bl/6, 2‐4 months) using a combination of filters with pore sizes of 300μm and 40μm followed by gradient centrifugation. BMVs were treated with FeTMPyP at 37oC for 60 minutes. Oxygen consumption rates (OCR) were measured using the Agilent Seahorse XFe24 analyzer and various respiratory parameters were determined following Mitostress test.ResultsIn male BMVs, basal respiration, ATP production, and non‐mitochondrial respiration were not altered by FeTMPyP treatment. Contrary to our hypothesis, in male BMVs, PN decomposition catalyst decreased the mitochondrial maximal respiration by 24.6% (2.1 ± 0.4 vs 2.8 ± 0.3 picomoles of O2/min/µg protein;) whereas the spare respiratory capacity was reduced by 33.3% (1.2 ± 0.3 vs 1.8 ± 0.3 picomoles of O2/min/µg protein; n=17 each, p<0.05). Proton leak was elevated by 70% (0.7 ± 0.1 vs 0.4 ± 0.1 picomoles of O2/min/µg protein) by PN decomposition catalyst in male BMVs. In contrast, in the female BMVs, the PN decomposition catalyst failed to alter mitochondrial respiratory parameters (n=15 each, p=NS). Interestingly, FeTMPyP increased non‐mitochondrial respiration by 63.8% (0.95 ± 0.2 vs 0.58 ± 0.1 picomoles of O2/min/µg protein) in the female BMVs (n=17 each, p<0.05).ConclusionBMVs display sex‐dependent respenses to endogenous PN. Notably, in female mouse BMVs, PN appears to act as an antioxidant as PN inhibited the non‐mitochondrial respiration which mostly contributes to extramitochondrial superoxide generation.