bluuebunny/crossref_metadata_2025 huggingface dataset:
Vasodilator mechanisms in intermediate‐ and small‐sized arteries from the toad, Bufo marinus (879.10)
Authors: Cameron Melissa , Donald John
Published: June 2021

Nitric oxide (NO), prostaglandins (PG) and endothelium‐derived hyperpolarising factor/s (EDHF) released by the endothelium are mediators of vasodilation in intermediate‐ and small‐sized resistance arteries of mammals. In this study, small vessel myography was used to determine the vasodilator mechanisms in the intermediate‐sized iliac artery and the small‐sized gastrocnemius artery of toad. Acetylcholine (ACh) was used to activate signalling systems that mediate vasodilation. In the iliac artery, ACh vasodilation was abolished by blockade of the NO and PG signalling pathways with ODQ and indomethacin, respectively. Furthermore, removal of the endothelium had no effect on the ACh vasodilation in the iliac artery. In contrast, ACh vasodilation in the gastrocnemius artery included NO and PG components, but there was a third endothelium‐dependent component that could be due to an EDHF, as the ACh dose‐response curve was shifted to the right by the K channel antagonist, TEA. Thus, the mechanisms of vasodilation in the amphibian arterial tree are dependent on the size of the vessel, and the endothelium may become more important as vessel size diminishes. This is the first time this has been shown in a non‐mammalian vertebrate in which the endothelial cells do not express an endothelial nitric oxide synthase.
Proteolytic cleavage of SPAK by a kidney protease (893.36)
Authors: Markadieu Nicolas , Spiller Benjamin , McDonald Hayes , Welling Paul , Delpire Eric
Published: June 2021

The Ste20 kinase SPAK is a major regulator of sodium reabsorption in the distal nephron. Recently, SPAK fragments have been detected in the mouse kidney, and particularly in the thick ascending limb of Henle (TAL). These truncated fragments have been demonstrated to inhibit the activity of the Na+‐K+‐2Cl‐ cotransporter, NKCC2, and thereby decrease sodium transport across the TAL. Here we show that mouse kidney lysates possess protease activity that leads to the specific cleavage of a SPAK, as we observed a precise and reproducible cleavage pattern with a GST‐SPAK fusion protein, but not a GST‐OSR1 fusion protein. This activity is not observed with brain, liver, or spleen lysates. The activity is insensitive to PMSF, leupeptin, aprotinin, and pepstatin, but inhibited by DTT and EDTA. In an effort to identify the protease, we performed ion exchange chromatography. The proteolytic activity was detected in an elution fraction corresponding to 250 mM NaCl. The fraction was subjected to MS/MS analysis and 1119 proteins were identified at 95% threshold. Among these proteins, we identified 44 peptidases and proteases. Gel permeation chromatography experiments determined that the protease is larger than 100 kDa. Effort will be made to determine the identity of the protein involved in the cleavage of SPAK and to identify the sites of cleavage.

Grant Funding Source: RO1 DK0935010
Functional characterization of a putative disaccharide membrane transporter in crustacean intestine (908.7)
Authors: Likely Rasheda , Ahearn Gregory
Published: June 2021

Mechanisms of transepithelial absorption of dietary sucrose in the American lobster, Homarus americanus, were investigated to determine if disaccharide sugars can be transported across an animal gut intact. Intestines were mounted in a perfusion chamber to measure mucosal to serosal (MS) 14C‐sucrose transport. MS sucrose transport was a hyperbolic function of luminal sugar concentration, suggesting the presence of carrier‐mediated transport. Sodium‐dependent glucose transport inhibitor, phloridzin, partially decreased MS 14C‐sucrose transport, suggesting hydrolysis of 14C‐sucrose to glucose during transport. Phloretin, an inhibitor of Na+‐independent basolateral glucose transport, partially decreased MS 14C‐sucrose transport, suggesting that some 14C‐sucrose radioactivity may be transported to the blood as 14C‐glucose. Decreased MS 14C‐sucrose transport also occurred in the presence of the disaccharide trehalose. Thin‐layer chromatography (TLC) was used to identify the chemical nature of radioactively‐labeled sugars in the bath following transport and revealed that 14C‐sucrose was transported across the intestine largely as an intact molecule. Results suggest that disaccharide sugars may be transported intact across crustacean intestine and provide support for the occurrence of a disaccharide membrane transporter that has not previously been functionally characterized.

Grant Funding Source: This project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2010‐65
Functional studies on the PKA regulatory subunit mutant R74C found in Carney complex (913.11)
Authors: Love Haley , Ogut Ozgur
Published: June 2021

The protein kinase A regulatory type I alpha subunit (PKA RegI) is the cAMP dependent subunit of the PKA holoenzyme. The gene for PKA RegI is located on chromosome 17q23‐17q24 and extends roughly 21 kb in length. PKA RegI is a component of the PKA holoenzyme, which is composed of a dimer of two regulatory subunits bound to a dimer of catalytic subunits (PKA Cat) to create a tetrameric protein in the inactive state. The PKA holoenzyme is activated by each PKA RegI monomer binding two molecules of cAMP at two distinct sites (3). This binding changes the shape of the regulatory subunit, therefore altering its affinity for the catalytic subunit by roughly 4 orders of magnitude (3). This causes the regulatory subunits to dissociate from the catalytic subunits, thereby releasing the catalytic subunits as active monomers to phosphorylate their intracellular targets. In their active state, the catalytic subunits act as active serine‐threonine kinases and have the potential to phosphorylate various targets which may alter muscle contractility as well as cell growth, proliferation and differentiation. PKA RegI is therefore an important protein with a vital role in the cardiovascular system, and its role in other tissues and disease states is being investigated. In particular, the Arg74Cys mutation of PKA RegI is found in Carney Complex, an autosomal dominant disorder that can cause a range of effects including skin lesions, tissue tumors (especially cardiac) and various types of endocrine tumors (1). It is therefore of interest to characterize the functional impact of this missense mutation on PKA RegI function in order to better understand the role of this mutation in the etiology of Carney Complex. Therefore, this study used biochemical and spectroscopic methods to examine the potential impact of the Arg74Cys mutation on the affinity of PKA RegI for both cAMP and its PKA Cat binding partner (2), as well as the effect of this mutation on PKA RegI phosphorylation at a novel downstream serine residue, Ser77.
Identification and characterization of the molecular interactions of persistent organic pollutants with the multidrug resistance transport protein ABCB1 (974.4)
Authors: Nicklisch Sascha , Bonito Lindsay , Sandin Stuart , Hamdoun Amro
Published: June 2021

Multidrug resistance (MDR) transporters, from the ATP binding cassette (ABC) superfamily, are thought to act as determinants of chemical accumulation in cells. Despite a long history of research on the interactions of MDR proteins with pharmaceuticals, little is known about the molecular basis of their interactions with environmental compounds such as persistent organic pollutants (POPs). This research is urgently needed to further understanding of substrate recognition by the transporters and to test the hypothesis that pollutant persistence relates to predictable patterns of interaction with the major MDR transporter ABCB1.

We use purified recombinant P‐glycoprotein (ABCB1) from mouse and yellowfin tuna to measure pollutant‐transporter interaction kinetics via ATPase stimulation and fluorophore knock‐off experiments. These kinetic will be compared to what is observed with ABCB1 overexpressed in whole cells. Previously analyzed data on “real‐world” pollutant levels in the cosmopolitan and widely consumed fish species of yellowfin tuna (Thunnus albacares) was used to guide the selection of persistent pollutants. The preliminary results of this study show that the majority of the selected persistent pollutants found in tuna are inhibitors of mouse ABCB1 efflux pump. Thereby, isomeric forms of the same pollutant can differ in their IC50 values up to an order of magnitude while reaching inhibition coefficients in the range of the model inhibitor cyclosporine A.

These studies aim at determining the conservation of pollutant‐transporter interactions across marine and mammalian ABCB1 transporters. In addition, we want to investigate whether common structural features of ABC transporters and pollutants govern their interactions. These results will shed light on the question of how transporters can direct pollutant movement through the environment and will lead to new avenues for design of more potent pharmaceuticals and safer industrial chemicals.

Grant Funding Source: Supported by NIH and the WAITT Foundation
Thermodynamic characterization of the interaction between the antimicrobial peptide LL‐37 and DNA oligonucleotides (978.3)
Authors: Craig Maria , Radford Michelle , Nguyen Tyler , Avelsgard Irmamarie
Published: June 2021

LL‐37 is a small peptide of innate immunity that sequesters extracellular DNA and transports it across the cell membrane. Recent discoveries have implicated LL‐37‐mediated pathways as potential inciting mechanisms for the onset of anti‐DNA autoimmunity. We have used electrophoretic mobility shift assays (EMSA), isothermal titration calorimetry (ITC), and western blotting to examine the details of complex formation between LL‐37 and 24‐bp DNA fragments with CpG, GpC, and poly(AT) sequences. ITC studies indicate that LL‐37 binding to CpG DNA fits a two‐site model, with association binding constants (Ka1 and Ka2) on the order of 108‐109 M‐1 and peptide:DNA ratios of 15‐25. Quantitative EMSA studies allow the construction of binding curves and demonstrate the formation of large peptide‐DNA complexes that form over a narrow concentration range. Parallel western blotting experiments detect LL‐37 self‐complexes that increase in size with increasing LL‐37 concentration and that decrease in size in the presence of DNA. LL‐37 appears to display higher affinity for poly(A/T) DNA than for CpG DNA in the EMSA assay. These data are consistent with a model in which LL‐37 self‐associations serve to tether DNA with high affinity into large complexes containing 20 LL‐37 molecules per 24 bp DNA fragment. This research was supported by the Mary Baldwin College Summer Undergraduate Research Fellows Program.
Poly‐ubiquitin chains paradox: development of novel and selective poly‐ubiquitin binding domains (LB184)
Authors: Brubaker Valerie , Loch Christian , Longenecker Kathryn , Strickler James
Published: June 2021

Ubiquitylation occurs through isopeptide linkage between the C‐terminus of Ub and the ε‐amino group of a lysine residue on the target substrate. Ub itself has seven Lys residues (K6, K11, K27, K29, K33, K48, and K63), each of which can participate in further ubiquitylation, generating poly‐Ub chains. The ability of Ub to form polymers through various lysines as well as the NH2‐terminus appears to be central to the versatility of this system in regulating cellular processes. The most extensively characterized of these polymers are linked through either K48 or K63. K48‐linked polyUb predominantly targets proteins for proteasomal degradation, whereas K63‐linked polyUb appears to regulate protein function, subcellular localization, or protein‐protein interactions. A growing body of evidence now implicates K11‐linked polyUb in mitotic regulation and endoplasmic reticulum associated degradation (ERAD). It is apparent from the above, that different poly‐ubiquitin linkages convey different information to the cell and suggests strongly that the cell contains elements capable of decoding this information, i.e. linkage specific ubiquitin binding domains. Development of Poly‐Ubiquitin selective tools will revolutionize the field of ubiquitin. Most UBDs described to date show little ability to discriminate between different linkages; although, it is possible to construct linkage specific tandem ubiquitin binding entities (TUBEs) by manipulating the spacing and rigidity of the linkers between tandem UBDs. We and our collaborators have constructed a series of K63‐specific UBDs and demonstrated their utility for Far Western blotting and “pull‐down” type experiments for evaluating the levels and identities of proteins bearing K63‐linked polyubiquitin chains under a variety of treatments. We will describe the results of proteomic experiments in which we identified new classes of UBDs. When validated, this information will have a major impact on our understanding of the functioning of the Ubiquitin‐Proteasome Pathway and suggest new targets for intervention.

Grant Funding Source: Supported by R43CA165561
RNA‐seq to identify novel markers for neural tissue differentiation (LB211)
Authors: Giorda Kristina , Sun Yongming , Frey Elizabeth , Taylor Madison , Barron Tori , Piper David , Meredith Gavin
Published: June 2021

Neural tissue differentiated and cultured from patient‐derived stem cells is expected to revolutionize the treatment of patients with brain and spinal injuries and diseases. Critical for these cellular therapies is accurate control and monitoring of differentiation but current methods for such cell typing are limited to qPCR and immunocytochemisty (ICC) which is not sufficient to discriminate between the numerous (likely >100,000) possible neural cell‐types. RNA(transcriptome aka RNA‐Seq) profiling permits the characterization and discovery of much‐needed novel markers. To define the temporal transcriptional signature of neural stem cells, cultured human embryonic stem cells (H9) were compared to induced neural stem cells (NSCs) at d0, d7 and d14. ICC was performed on the putative NSC pools at d7 and d14 for markers of pluripotency (Oct4) and neural differentiation (nestin, Sox1, and Pax6) and H9 cells were stained on d14 for markers of pluripotency (Oct4 and SSEA4). Total RNA was isolated over the time course from the undifferentiated and differentiated cells. Ion Torrent libraries were created to profile expression of miRNAs and whole transcriptomes for each cell population. Multiplexed Proton sequencing and Torrent Suite Software analysis yielded 蠅2.5 million small RNA reads and 蠅29 million whole transcriptome reads per sample. Cluster analysis of the RNA‐Seq profiles indicates that the cell populations have characteristic molecular signatures. Among genes that are decreased in induced cells are OCT4 (POU5F1), JARID2, NANOG , consistent with the differentiation of iPSCs into neurons. Among genes that showed increased expressions are NTRK2, POU3F2, and a number of HOX family genes. We also find lincRNA are involved in cell differentiation.
AMPK and PTP‐1B stand at the gates of immunity, metabolism and egg formation in mosquitoes. (LB224)
Authors: Silva‐Neto Mario Alberto , Nunes Rodrigo , Moretti Debora , Cudischevitch Cecilia , Daumas‐Filho Carlos Renato , Medeiros‐Castro Priscilla , Jablonka Willy , Ventura‐Martins Guilherme , Capurro Margareth , Mesquita Rafael , Ahuja Lalima , Atella Georgia
Published: June 2021

Mosquitoes are major vectors of human diseases. This is mainly due to their requirement for a blood meal to lay eggs which allows the transmission of pathogens to the human being. Our aim is to identify proper molecular targets in order to block blood feeding and impair egg formation.

In their early days of adult life mosquitoes feed on vegetal fluids and their midgut is continuously loaded with several bioactive compounds including polyphenols. We observed that polyphenol‐fed mosquitos dramatically increase their average lifespan (Control T 1/2 ± 15‐18 days, Polyphenol‐fed T1/2± 20‐32 days). Polyphenols also reduced the body triglyceride content from 20‐75 %. They trigger midgut autophagy which leads to a 40 % decrease on bacterial loads. Such mechanisms are mimicked by the activation of the mosquito AMPK and abolished by RNAi‐mediated silencing of this gene. Thus the presence of polyphenol on early mosquito meals optimizes mosquito metabolism and immunity and its chances of finding its first blood meal. After a blood meal digestion will take place in mosquito midgut. Blood‐derived nutrients allow yolk synthesis and egg formation. Blood feeding affects the expression of the mosquito PTP‐1B (AaPTP1B) in tissues critically involved in the synthesis and storage of vitellogenin (Vg), the major yolk protein. The protein tyrosine phosphatase inhibitor sodium orthovanadate or the PTP substrate DiFMUP decreased Vg synthesis and egg production. Also, silencing of AaPTP1B using by RNAi assays suppressed by 30 % egg production.

In conclusion, the above results show that mosquito signaling pathways regulated by AMPK and PTP1B are proper targets to block blood feeding and egg formation. Future research strategies should be designed to generate novel cell signaling‐based insecticides that simultaneously decrease mosquito lifespan and reproduction.

Grant Funding Source: Supported by CNPq, INCT‐EM, FAPERJ
Using the Amira software to Reconstruct a 3D Kidney Model (LB25)
Authors: Pavlov Anna , Chen Jian , Yang Tong , Lynch James , Cui Dongmei
Published: June 2021

Amira is a powerful software for bioscience research that anatomists can use to create 3D structures from CT, MRI, and other diagnostic images. Amira allows us to view 3‐dimensional structures not only from outside but also from the inside of the kidney. In this project, we reconstructed the right‐sided kidney through an ultra‐thin MRI. The structures of the cortex, medulla, calyces, urethra, and renal artery and vein were assigned pseudocolor to simplify identification. We demonstrated how Amira can be used by our students to view multiple displays of 3D kidney models and internal viewing of the cortex and medulla. Amira carries a handful of tools for easily reconstructing CT/MRI images into 3D structures, saving time from tedious segmental outlining and giving a better and more detailed view of the kidney in cross‐section.

Grant Funding Source: UMC
High molecular weight kininogen binds phosphatidylserine and opsonizes urokinase plasminogen activator receptor‐mediated efferocytosis (LB529)
Authors: Wu Yi
Published: June 2021

Phagocytosis of apoptotic cells (efferocytosis) is essential for regulation of immune responses and tissue homeostasis, and is mediated by specific receptors on phagocytes. In this study we found that urokinase plasminogen activator receptor (uPAR) plays an important role in internalization of apoptotic cells, and also characterized the underlying mechanisms. In a flow cytometry‐based phagocytic assay, uPAR‐deficient (uPAR‐/‐) macrophages displayed significant defect in internalization but not tethering of apoptotic cells. When uPAR‐/‐ mice were challenged with apoptotic cells, they exhibited pronounced splenomegaly resulting from accumulation of abundant apoptotic cells in spleen. Overexpression of uPAR in HEK‐293 cells enhanced efferocytosis, which was inhibited by annexin V and phosphatidylserine (PS) liposome, suggesting that uPAR‐mediated efferocytosis is dependent on PS. In serum lacking high‐molecular‐weight kininogen (HK), a uPAR ligand, uPAR‐mediated efferocytosis was significantly attenuated, which was rescued by replenishment of HK. As detected by flow cytometry, HK selectively bound to apoptotic cells, but not viable cells. In purified systems, HK was specifically associated with PS liposome. HK binding to apoptotic cells induced its rapid cleavage to two‐chain HKa and bradykinin. Both heavy chain and light chain of HKa were associated with PS liposome and apoptotic cells. HKa has higher binding affinity than HK to uPAR. Overexpression of Rac1/N17 cDNA inhibited uPAR‐mediated efferocytosis. HK plus PS liposome stimulated a complex formation of CrkII with p130Cas and Dock‐180, and Rac1 activation in uPAR‐293 cells, not in 293 cells. Thus, uPAR mediates efferocytosis through HK interaction with PS on apoptotic cells and activation of Rac1 pathway.
Age‐Related Changes Heart Function in the Female SHR, a Model of Postmenopausal Cardiovascular Physiology (LB712)
Authors: McCarthy Joseph , Ramirez Rolando , Novak Jacqueline
Published: June 2021

Age‐Related Changes Heart Function in the Female SHR, a Model of Postmenopausal Cardiovascular Physiology. Joseph McCarthy1, Rolando J.J. Ramirez1, Jacqueline Novak2. 1Biology, The University of Akron, Akron, OH, 2Department of Math and Sciences, Walsh University, N. Canton, OH.

Women have a decreased risk of cardiovascular disease compared to men until after menopause when the risk is similar between men and women. The mechanism for cardioprotective effect is unclear. In order to study the physiological changes which occur in the cardiovascular system of women after menopause, it is necessary to identify animal models. One model of post‐menopausal physiology is the aged spontaneously hypertensive female rats (SHR). The endocrine and blood pressure changes have been previously recorded by Reckelhoff and colleagues; however, the cardiac function has not been examined via isolated heart techniques. The purpose of this study was to compare the contractility of hearts isolated from postmenopausal rats (aged SHR females) compared to young SHR controls. . Hearts studied using an isolated Langendorff system. The contractility measurements were recorded at 3 different ventricular volumes (0.05, 0.10, and 0.15ml) using a pressure transducer and the data were collected using WinDaq (DI‐720‐Di‐7x0). Contractility was calculated by the change in pressure over the change in time(dP/dt). A significant increase in contractility was shown between the young and old SHR models at the three different volumes. For example, at 0.15ml, the contractility was 6.19 1.49 in the post‐menopausal rats compared to 4.03 0.91 in the young SHR. These results indicate that there are significant age‐related changes in the isolated heart of aged SHR which is further evidence that this is an important model for studying post‐menopausal changes in heart function.

Grant Funding Source: Supported by NIH 1R15HL097343‐01A2
Use of Soft Contact Lens for Esthetic and Functional Rehabilitation
Authors: de Paula Yoneda Patrícia , Schellini Silvana A. , Padovani Carlos R. , Silva Virgílio F.
Published: June 2021

Objective:

To report the outcomes and complications of colored soft contact lenses (SCLs) to improve esthetic or eye function in visually impaired (VI) or blind patients.

Methods:

This retrospective study enrolled consecutive patients with unilateral or bilateral VI or blindness who had received colored SCLs to improve appearance or ocular function. Data were collected on demographics, complaints, complete ophthalmological examinations, types of SCL, follow-ups, and complications.

Results:

The study sample was comprised of 109 patients (ages 4–80 years). Eighty-six patients (78.89%) had unilateral blindness, and 14 patients (12.84%) had functional disorder. Most of the patients presenting for improved esthetic had anterior segment pathology (74/67.88%), mainly secondary to trauma (55/50.5%) resulting in leukoma (73/66.97%). Patients with functional VI (19/17.43%) received filtered SCLs for color blindness (7/6.42%), diplopia (7/6.42%), or photophobia (5/4.58%). No serious complications were observed in any cases, and 94.49% of patients were satisfied with the SCLs.

Conclusion:

Colored SCLs can cover ocular surface imperfections in VI or blind eyes of patients who are seeking better cosmesis or improvement functional VI. Soft contact lenses with only one filter can cover imperfections of the ocular surface and improve vision in functional VI. Appropriate follow-up of these patients can reduce complication rates, resulting in good cosmesis and patient satisfaction with a positive impact on the lives of patients suffering from functional VI or blindness.
Risk factors for antibody formation in children with hemophilia: methodological aspects and clinical characteristics of the HEMFIL cohort study
Authors: Jardim Letícia L. , Santana Marcio P. , Chaves Daniel G. , van der Bom Johanna , Rezende Suely M.
Published: June 2021

Up to 35% of patients with hemophilia A and 5% with hemophilia B develop neutralizing antibodies which can inhibit the therapeutic activity of factor replacement (inhibitors). Despite the clinical relevance of antifactor VIII and IX neutralizing antibodies, there is still a major gap on the knowledge of risk factors for their development. Furthermore, most of the studies on risk factors for inhibitor development come from Caucasian and Afro-American populations. The HEMFIL is a Brazilian prospective cohort study of previously untreated children with hemophilia, which primary aim is to identify new risk factors related to inhibitor development. This manuscript aims at describing the study design and its methodology. After the diagnosis, children are followed up to 75 exposure days or to inhibitor development. Standardized forms and blood samples are collected to describe clinical characteristics and to perform the measurement of immunological and genetic biomarkers at three time points; Inclusion time (T0), at inhibitor development or at 75 exposure days without inhibitors (T1) and after immune tolerance induction for patients in whom it is indicated and performed (T2). Currently, 120 children have been included, of whom, 95 have completed the follow-up. For severe/moderately severe hemophilia A, the cumulative incidence of inhibitors at 75 exposure days was 35% (95% confidence interval, 26–46%). The inclusion of additional patients and a longer follow-up will allow the analysis of risk factors for inhibitor development.
Validation of the BPro radial pulse waveform acquisition device in pregnancy and gestational hypertensive disorders
Authors: Jakes Adam , Wade Julie , Vowles Zoe , Seed Paul T. , Shennan Andrew H. , Chappell Lucy C. , Nzelu Diane
Published: June 2021

Objective

To validate the BPro blood pressure (BP) wrist device for use in pregnancy and preeclampsia according to the Universal Standard protocol.

Participants and methods

BP was measured sequentially in 45 pregnant women (including 15 with preeclampsia, 15 with gestational hypertension and 15 who remained normotensive) alternating between a mercury sphygmomanometer and BPro device.

Results

The BPro is accurate in pregnancy with a mean device-observer difference of −1.7 ± 6.1 and 0.1 ± 4.6 mmHg for SBP and DBP, respectively. In women with preeclampsia, BPro also met the validation criteria for the Universal Standard protocol with a mean device-observer difference of −2.7 ± 7.1 and 0.3 ± 4.7 mmHg for SBP and DBP, respectively. However, the number of absolute BP differences within 5 mmHg was considerably fewer in those with preeclampsia when compared to the other two subgroups.

Conclusion

The BPro device can be recommended for BP measurement in pregnancy but should be used with caution in those with confirmed preeclampsia.
A novel method for the noninvasive estimation of cardiac output with brachial oscillometric blood pressure measurements through an assessment of arterial compliance
Authors: Álvarez-Montoya Diego , Madrid-Muñoz Camilo , Escobar-Robledo Luis , Gallo-Villegas Jaime , Aristizábal-Ocampo Dagnovar
Published: June 2021

Objective

To propose and validate a new method for estimating cardiac output based on the total arterial compliance (Ct) formula that does not need an arterial waveform and to apply it to brachial oscillometric blood pressure measurements (OBPMs).

Methods

One hundred subjects with normal heart anatomy and function were included. Reference values for cardiac output were measured with echocardiography, and Ctwas calculated with a two-element Windkessel model. Then, a statistical model of arterial compliance (Ce) was used to estimate cardiac output. Finally, the measured and estimated cardiac output values were compared for accuracy and reproducibility.

Results

The model was derived from the data of 70 subjects and prospectively tested with the data from the remaining 30 individuals. The mean age of the whole group was 43.4 ± 12.8 years, with 46% women. The average blood pressure (BP) was 107.1/65.0 ± 15.0/9.6 mmHg and the average heart rate was 67.7 ± 11.4 beats/min. The average Ctwas 1.39 ± 0.27 mL/mmHg and the average cardiac output was 5.5 ± 1.0 L/min. The mean difference in the cardiac output estimated by the proposed methodology vs. that measured by Doppler echocardiography was 0.022 L/min with an SD of 0.626 L/min. The intraclass correlation coefficient was 0.93, and the percentage error was 19%.

Conclusion

Cardiac output could be reliably and noninvasively obtained with brachial OBPMs through a novel method for estimating Ctwithout the need for an arterial waveform. The new method could identify hemodynamic factors that explain BP values in an ambulatory care setting.
Incident hyperuricemia in relation to antihypertensive therapy with the irbesartan/hydrochlorothiazide combination
Authors: Zhang Di , Huang Qi-Fang , Li Yan , Wang Ji-Guang
Published: June 2021

Objective

We investigated serum uric acid changes and incident hyperuricemia in relation to the achieved blood pressure (BP) after 12 weeks of antihypertensive therapy with the irbesartan/hydrochlorothiazide combination.

Methods

The study participants were 449 patients who completed the study. Analysis of covariance and multiple logistic regression analyses were performed to calculate the least square mean changes (± standard error) from baseline in serum uric acid and odds ratios (ORs) for incident hyperuricemia according to the achieved levels of BP.

Results

Adjusted analyses showed that serum uric acid changes differed according to the achieved SBP/DBP (P = 0.002), with a smaller mean (± standard error) increase in the range of 130–139/<90 mm Hg (n = 132, 19.8 ± 5.7 µmol/L) than that of ≥140/90 (n = 129, 32.4 ± 7.3 µmol/L) or <130/90 mm Hg (n = 188, 39.5 ± 5.1 µmol/L). Adjusted analyses showed similar results for the incident hyperuricemia (n = 95, 24.0%) in those patients with normal serum uric acid at baseline (n = 396). The risk of incident hyperuricemia was lower (OR, 0.45; 95% confidence interval 0.25–0.83; P = 0.04) in patients with an achieved SBP/DBP of 130–139/<90 mm Hg (n = 117, incidence rate, 17.1%) than those with an achieved SBP/DBP of ≥140/90 (n = 118, 31.4%) or <130/90 mm Hg (n = 161, 23.6%).

Conclusions

Thiazide-induced changes in serum uric acid or incident hyperuricemia were associated with the achieved SBP/DBP, being lower at the level of 130–139/<90 mm Hg.
The Genitourinary Pathology Society Update on Classification and Grading of Flat and Papillary Urothelial Neoplasia With New Reporting Recommendations and Approach to Lesions With Mixed and Early Patterns of Neoplasia
Authors: Amin Mahul B. , Comperat Eva , Epstein Jonathan I. , True Lawrence D. , Hansel Donna , Paner Gladell P. , Al-Ahmadie Hikmat , Baydar Dilek , Bivalacqua Trinity , Brimo Fadi , Cheng Liang , Cheville John , Dalbagni Guido , Falzarano Sara , Gordetsky Jennifer , Guo Charles C. , Gupta Sounak , Hes Ondra , Iyer Gopa , Kaushal Seema , Kunju Lakshmi , Magi-Galluzzi Cristina , Matoso Andres , Netto George , Osunkoya Adeboye O. , Pan Chin Chen , Pivovarcikova Kristina , Raspollini Maria R. , Reis Henning , Rosenberg Jonathan , Roupret Morgan , Shah Rajal B. , Shariat Shahrokh , Trpkov Kiril , Weyerer Veronika , Zhou Ming , McKenney Jesse , Reuter Victor E.
Published: June 2021

The Genitourinary Pathology Society (GUPS) undertook a critical review of the recent advances in bladder neoplasia with a focus on issues relevant to the practicing surgical pathologist for the understanding and effective reporting of bladder cancer, emphasizing particularly on the newly accumulated evidence post-2016 World Health Organization (WHO) classification. The work is presented in 2 manuscripts. Here, in the first, we revisit the nomenclature and classification system used for grading flat and papillary urothelial lesions centering on clinical relevance, and on dilemmas related to application in routine reporting. As patients of noninvasive bladder cancer frequently undergo cystoscopy and biopsy in their typically prolonged clinical course and for surveillance of disease, we discuss morphologies presented in these scenarios which may not have readily applicable diagnostic terms in the WHO classification. The topic of inverted patterns in urothelial neoplasia, particularly when prominent or exclusive, and beyond inverted papilloma has not been addressed formally in the WHO classification. Herein we provide a through review and suggest guidelines for when and how to report such lesions. In promulgating these GUPS recommendations, we aim to provide clarity on the clinical application of these not so uncommon diagnostically challenging situations encountered in routine practice, while also importantly advocating consistent terminology which would inform future work.
The Genitourinary Pathology Society Update on Classification of Variant Histologies, T1 Substaging, Molecular Taxonomy, and Immunotherapy and PD-L1 Testing Implications of Urothelial Cancers
Authors: Compérat Eva , Amin Mahul B. , Epstein Jonathan I. , Hansel Donna E. , Paner Gladell , Al-Ahmadie Hikmat , True Larry , Bayder Dilek , Bivalacqua Trinity , Brimo Fadi , Cheng Liang , Cheville John , Dalbagni Guido , Falzarano Sara , Gordetsky Jennifer , Guo Charles , Gupta Sounak , Hes Ondrej , Iyer Gopa , Kaushal Seema , Kunju Lakshmi , Magi-Galluzzi Cristina , Matoso Andres , McKenney Jesse , Netto George J. , Osunkoya Adeboye O. , Pan Chin Chen , Pivovarcikova Kristina , Raspollini Maria R. , Reis Henning , Rosenberg Jonathan , Roupret Morgan , Shah Rajal B. , Shariat Shahrokh F. , Trpkov Kiril , Weyerer Veronika , Zhou Ming , Reuter Victor
Published: June 2021

The Genitourinary Pathology Society (GUPS) undertook a critical review of the recent advances in bladder cancer focusing on important topics of high interest for the practicing surgical pathologist and urologist. This review represents the second of 2 manuscripts ensuing from this effort. Herein, we address the effective reporting of bladder cancer, focusing particularly on newly published data since the last 2016 World Health Organization (WHO) classification. In addition, this review focuses on the importance of reporting bladder cancer with divergent differentiation and variant (subtypes of urothelial carcinoma) histologies and the potential impact on patient care. We provide new recommendations for reporting pT1 staging in diagnostic pathology. Furthermore, we explore molecular evolution and classification, emphasizing aspects that impact the understanding of important concepts relevant to reporting and management of patients.
Fractionally integrated Bessel process
Authors: Shevchenko Georgiy , Zatula Dmytro
Published: June 2021

We consider a fractionally integrated Bessel process defined by

Y
s

δ
,
H

=

∫
0
∞

(

u

H
−
(

1

/

2

)

−

(
u
−
s
)

+

H
−
(

1

/

2

)

)

d

X
u
δ

, where

X
δ

is the Bessel process of dimension
δ
 > 2. We discuss the relation of this process to the fractional Brownian motion at its maximum, study the basic properties of the process and prove its Hölder continuity.
Dislocation dynamics modelling of the creep behaviour of particle-strengthened materials
Authors: liu F. X. , Cocks A. C. F , Tarleton E.
Published: June 2021

Plastic deformation in crystalline materials occurs through dislocation slip and strengthening is achieved with obstacles that hinder the motion of dislocations. At relatively low temperatures, dislocations bypass the particles by Orowan looping, particle shearing, cross-slip or a combination of these mechanisms. At elevated temperatures, atomic diffusivity becomes appreciable, so that dislocations can bypass the particles by climb processes. Climb plays a crucial role in the long-term durability or creep resistance of many structural materials, particularly under extreme conditions of load, temperature and radiation. Here we systematically examine dislocation-particle interaction mechanisms. The analysis is based on three-dimensional discrete dislocation dynamics simulations incorporating impenetrable particles, elastic interactions, dislocation self-climb, cross-slip and glide. The core diffusion dominated dislocation self-climb process is modelled based on a variational principle for the evolution of microstructures, and is coupled with dislocation glide and cross-slip by an adaptive time-stepping scheme to bridge the time scale separation. The stress field caused by particles is implemented based on the particle–matrix mismatch. This model is helpful for understanding the fundamental particle bypass mechanisms and clarifying the effects of dislocation glide, climb and cross-slip on creep deformation.
Insights into evolution and coexistence of the colibactin- and yersiniabactin secondary metabolite determinants in enterobacterial populations
Authors: Wami Haleluya , Wallenstein Alexander , Sauer Daniel , Stoll Monika , von Bünau Rudolf , Oswald Eric , Müller Rolf , Dobrindt Ulrich
Published: June 2021

The bacterial genotoxin colibactin interferes with the eukaryotic cell cycle by causing dsDNA breaks. It has been linked to bacterially induced colorectal cancer in humans. Colibactin is encoded by a 54 kb genomic region in

Enterobacteriaceae

. The colibactin genes commonly co-occur with the yersiniabactin biosynthetic determinant. Investigating the prevalence and sequence diversity of the colibactin determinant and its linkage to the yersiniabactin operon in prokaryotic genomes, we discovered mainly species-specific lineages of the colibactin determinant and classified three main structural settings of the colibactin–yersiniabactin genomic region in

Enterobacteriaceae

. The colibactin gene cluster has a similar but not identical evolutionary track to that of the yersiniabactin operon. Both determinants could have been acquired on several occasions and/or exchanged independently between enterobacteria by horizontal gene transfer. Integrative and conjugative elements play(ed) a central role in the evolution and structural diversity of the colibactin–yersiniabactin genomic region. Addition of an activating and regulating module (clbAR) to the biosynthesis and transport module (clbB-S) represents the most recent step in the evolution of the colibactin determinant. In a first attempt to correlate colibactin expression with individual lineages of colibactin determinants and different bacterial genetic backgrounds, we compared colibactin expression of selected enterobacterial isolates in vitro. Colibactin production in the tested

Klebsiella

species and

Citrobacter koseri

strains was more homogeneous and generally higher than that in most of the

Escherichia coli

isolates studied. Our results improve the understanding of the diversity of colibactin determinants and its expression level, and may contribute to risk assessment of colibactin-producing enterobacteria.
Genomic contextualisation of ancient DNA molecular data from an Argentinian fifth pandemic Vibrio cholerae infection
Authors: Dorman Matthew J. , Thomson Nicholas R. , Campos Josefina
Published: June 2021

Specific lineages of serogroup O1

Vibrio cholerae

are notorious for causing cholera pandemics, of which there have been seven since the 1800s. Much is known about the sixth pandemic (1899–1923) and the ongoing seventh pandemic (1961–present), but we know very little about the bacteriology of pandemics 1 to 5. Moreover, although we are learning about the contribution of non-O1 non-pandemic

V. cholerae

to cholera dynamics during the current pandemic, we know almost nothing about their role in the past. A recent ancient DNA study has presented what may be the first molecular evidence of a

V. cholerae

infection from the fifth cholera pandemic period (1886–1887 AD) in Argentina. Here, we place the molecular evidence from that study into the genomic context of non-pandemic

V. cholerae

from Latin America and elsewhere, and show that a gene fragment amplified from ancient DNA is most similar to that of

V. cholerae

from the Americas, and from Argentina. Our results corroborate and reinforce the findings of the original study, and collectively suggest that even in the 1880s, non-pandemic

V. cholerae

local to the Americas may have caused sporadic infections in Argentina, just as we know this to have happened during the seventh pandemic in Latin America.
Patient Outcomes and Lessons-Learned from Treating Patients with Severe COVID-19 at a Long-Term Acute Care Hospital
Authors: Grevelding Pete , Hrdlicka Henry C. , Holland Stephen , Cullen Lorraine , Meyer Amanda , Connors Catherine , Cooper Darielle , Greco Allison
Published: June 2021

Objective

To describe characteristics, clinical management, and patient outcomes during and after acute COVID-19 phase in a long-term acute care hospital in the Northeastern United States.

Methods

A single-center retrospective analysis of electronic medical records of patients treated for COVID-19-related impairments, from March 19, 2020 through August 14, 2020, was conducted to evaluate patient outcomes in response to the facility’s holistic treatment approach.

Results

118 admissions were discharged by the data cut-off. Mean patient age was 63 years, 64.1% were male, and 29.9% of patients tested-positive for SARS-CoV-2 infection at admission. The mean (SD) length-of-stay at was 25.5 (13.0) days and there was a positive correlation between patient age and length-of-stay. Of the 51 patients non-ambulatory at admission, 83.3% were ambulatory at discharge. Gait increased 217.4 feet from admission to discharge, a greater increase than the reference cohort of 146.3 feet. 93.8% (15/16) of patients mechanically ventilated at admission were weaned before discharge (mean 11.3 days). 74.7% (56/75) of patients admitted with a restricted diet were discharged on a regular diet.

Conclusion

The majority of patients treated at a long-term acute care hospital for severe COVID-19 and related complications improved significantly through coordinated care and rehabilitation.
SIGMA METRICS ASSESSMENT AS QUALITY IMPROVEMENT METHODOLOGY IN A CLINICAL CHEMISTRY LABORATORY
Authors: Garg Monika , Sharma Neera , Das Saswati
Published: June 2021

Background

The concept of sigma metrics & lean six sigma is well known in the field of healthcare. However not many labs utilize the six sigma metrics for maintenance of high quality laboratory performance. A minimum value of 3 σ is desired in any clinical laboratory & values of σ≥ 6 are regarded as gold standard for obtaining high quality lab reports.

Aims &Objectives

To calculate bias, cv & sigma metrics from the IQC & EQC data in order to ascertain extent of quality management in our lab.

Materials &Methods

An extensive study of sample processing and quality practices was carried out in the Central Laboratory of Department of Biochemistry; PGIMER &Dr. RML Hospital, New Delhi; from Feb 2020 to July 2020. The IQC used(both level I & level II) were from Biorad Laboratories India (lyphochek assayed chemistry control) & the EQC used was from Randox Laboratories, UK. All the controls were run on Beckman Coulter clinical chemistry analyser AU 680. Total 14 clinical parameters were analysed & subsequently; Mean, S.D., CV, bias & σ were calculated through their respective formulas.

Results

Sigma level was more than 6 for both levels of IQC was observed for Amylase. It indicates world class performance. Total bilirubin, AST, Triglyceride & HDL depicted σ values between 3.1 – 6 for both L1& L2.Iron showed σ value of 5.5 in L1whereas it was 3.78 in L2.

Conclusion

Sigma metrics in clinical laboratory is an essential technique to ascertain poor assay performance, along with assessment of the efficiency of existing laboratory process.
A large disordered region confers a wide spanning volume to vertebrate Suppressor of Fused as shown in a trans-species solution study
Authors: Makamte Staëlle , Jabrani Amira , Paquelin Annick , Plessis Anne , Sanial Mathieu , Thureau Aurélien , Rudenko Olga , Oteri Francesco , Baaden Marc , Biou Valérie
Published: June 2021

Hedgehog (Hh) pathway inhibition by the conserved protein Suppressor of Fused (SuFu) is crucial to vertebrate development. By constrast, SuFu removal has little effect in drosophila.

Previous publications showed that the crystal structures of human and drosophila SuFu consist of two ordered domains that are capable of breathing motions upon ligand binding. However, the crystal structure of human SuFu does not give information about 20 N-terminal residues (IDR1) and an eighty-residue-long disordered region (IDR2) in the C-terminus, whose function is important for the pathway repression. These two IDRs are species-dependent.

We studied SuFu’s structure in solution, both with circular dichroism and small angle X-ray scattering, comparing drosophila, zebrafish and human species, to better understand this considerable difference. Our studies show that, in spite of similar crystal structures restricted to ordered domains, drosophila and vertebrate SuFu have very different structures in solution. The IDR2 of vertebrates spans a large area, thus enabling it to reach for partners and be accessible for post-translational modifications. Furthermore, we show that the IDR2 region is highly conserved within phyla but varies in length and sequence, with insects having a shorter disordered region while that of vertebrates is broad and mobile. This major variation may explain the different phenotypes observed upon SuFu removal.
Willingness to Pay in the Human Brain: A fMRI Activation Likelihood Estimation Meta-analysis
Authors: Yao Xiaoqiang , Huang Zhigang , Wang Yiwen
Published: June 2021

The neural substrate of willingness to pay (WTP) ultimately supports human economic exchange activities and plays a crucial role in daily life. This paper aimed to identify the neural basis of WTP for food and nonfood, as well as the brain regions related to real and hypothetical WTP choices. We found the human brain centers of WTP by performing an activation likelihood estimation (ALE) meta-analysis (27 experiments, 796 subjects) on the existing neuroimaging studies. The conjunction analysis revealed that WTP for food and nonfood engaged a common cluster in the paracingulate and cingulate gyrus, revealing a common reward circuit in the brain. The frontal medial cortex and paracingulate gyrus were particularly activated by WTP for nonfood. Furthermore, the left caudate, left thalamus, angular gyrus and supramarginal gyrus (subregions of inferior parietal lobule) were more convergently activated by hypothetical WTP choice. Our findings support the idea that a common currency representation in the brain and reward-specific neural basis. Results also provide evidence of neural representations of the hypothetical bias.