bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of Infrared Thermography for Assessment of Burn Depth and Healing Potential: A Systematic Review
Authors: Dang Justin , Lin Matthew , Tan Calvin , Pham Christopher H , Huang Samantha , Hulsebos Ian F , Yenikomshian Haig , Gillenwater Justin
Published: June 2021

Burn wound depth assessments are an important component of determining patient prognosis and making appropriate management decisions. Clinical appraisal of the burn wound by an experienced burn surgeon is standard of care but has limitations. Infrared (IR) thermography is a technology in burn care that can provide a noninvasive, quantitative method of evaluating burn wound depth. IR thermography utilizes a specialized camera that can capture the IR emissivity of the skin, and the resulting images can be analyzed to determine burn depth and healing potential of a burn wound. Though IR thermography has great potential for burn wound assessment, its use for this has not been well documented. Thus, we have conducted a systematic review of the current use of IR thermography to assess burn depth and healing potential. A systematic review and meta-analysis of the literature was performed on PubMed and Google Scholar between June 2020 and December 2020 using the following keywords: FLIR, FLIR ONE, thermography, forward looking infrared, thermal imaging + burn*, burn wound assessment, burn depth, burn wound depth, burn depth assessment, healing potential, burn healing potential. A meta-analysis was performed on the mean sensitivity and specificity of the ability of IR thermography for predicting healing potential. Inclusion criteria were articles investigating the use of IR thermography for burn wound assessments in adults and pediatric patients. Reviews and non-English articles were excluded. A total of 19 articles were included in the final review. Statistically significant correlations were found between IR thermography and laser Doppler imaging (LDI) in 4/4 clinical studies. A case report of a single patient found that IR thermography was more accurate than LDI for assessing burn depth. Five articles investigated the ability of IR thermography to predict healing time, with four reporting statistically significant results. Temperature differences between burnt and unburnt skin were found in 2/2 articles. IR thermography was compared to clinical assessment in five articles, with varying results regarding accuracy of clinical assessment compared to thermography. Mean sensitivity and specificity of the ability of IR thermography to determine healing potential <15 days was 44.5 and 98.8, respectively. Mean sensitivity and specificity of the ability of FLIR to determine healing potential <21 days was 51.2 and 77.9, respectively. IR thermography is an accurate, simple, and cost-effective method of burn wound assessment. FLIR has been demonstrated to have significant correlations with other methods of assessing burns such as LDI and can be utilized to accurately assess burn depth and healing potential. However, the technology needs more standardization and validation before it can achieve widespread adoption in clinical practice.
Virulence of Entomopathogenic Nematodes to Pupae ofFrankliniella fusca(Thysanoptera: Thripidae)
Authors: Gulzar Sehrish , Usman Muhammad , Wakil Waqas , Wu Shaohui , Oliveira-Hofman Camila , Srinivasan Rajagopalbabu , Toews Michael , Shapiro-Ilan David
Published: June 2021

Tobacco thrips, Frankliniella fusca (Hinds) is an economically significant pest. Entomopathogenic nematodes (EPNs) have shown promise as biocontrol agents against certain thrips species, but they have not been explored for suppression of F. fusca. We investigated the potential of EPNs to manage F. fusca by conducting three different bioassays: 1) a small cup dose–response bioassay (25, 50, and 100 IJs cm−2) with four EPN species, 2) a broad virulence bioassay with eight EPN species at 100 IJs cm−2, and 3) a potted soil bioassay testing with four EPN species (100 IJs cm−2). In the dose–response bioassay, all treatments showed relatively lower adult emergence when compared with the control group, but the minimum adult emergence (30%) was observed at 7 d post-treatment when Heterorhabditis bacteriophora (FL1-1) was applied at the highest rate (100 IJs cm−2). In the broad virulence study, all EPN treatments caused significant reductions in F. fusca adult emergence (18.3–75.0%) in comparison with the control. H. bacteriophora (Fl1-1) was more virulent than other nematode treatments but statistically not different from Steinernema feltiae and Steinernema riobrave, while Steinernema rarum was the least virulent. In the potted soil bioassay, the lowest emergence (10.6%) was observed in H. bacteriophora (Fl1-1) treatment, followed by S. feltiae (SN), S. riobrave (355), and Heterorhabditis indica (HOM1) treatments. These results indicate that EPNs have the ability to suppress the soil dwelling stage of F. fusca and should be explored further under greenhouse and field conditions for biocontrol potential within an integrated pest management (IPM) context.
Association Between Smoking and Molecular Subtypes of Colorectal Cancer
Authors: Wang Xiaoliang , Amitay Efrat , Harrison Tabitha A , Banbury Barbara L , Berndt Sonja I , Brenner Hermann , Buchanan Daniel D , Campbell Peter T , Cao Yin , Chan Andrew T , Chang-Claude Jenny , Gallinger Steven J , Giannakis Marios , Giles Graham G , Gunter Marc J , Hopper John L , Jenkins Mark A , Lin Yi , Moreno Victor , Nishihara Reiko , Newcomb Polly A , Ogino Shuji , Phipps Amanda I , Sakoda Lori C , Schoen Robert E , Slattery Martha L , Song Mingyang , Sun Wei , Thibodeau Steven N , Toland Amanda E , Van Guelpen Bethany , Woods Michael O , Hsu Li , Hoffmeister Michael , Peters Ulrike
Published: June 2021

Background

Smoking is associated with colorectal cancer (CRC) risk. Previous studies suggested this association may be restricted to certain molecular subtypes of CRC, but large-scale comprehensive analysis is lacking.

Methods

A total of 9789 CRC cases and 11 231 controls of European ancestry from 11 observational studies were included. We harmonized smoking variables across studies and derived sex study–specific quartiles of pack-years of smoking for analysis. Four somatic colorectal tumor markers were assessed individually and in combination, including BRAF mutation, KRAS mutation, CpG island methylator phenotype (CIMP), and microsatellite instability (MSI) status. A multinomial logistic regression analysis was used to assess the association between smoking and risk of CRC subtypes by molecular characteristics, adjusting for age, sex, and study. All statistical tests were 2-sided and adjusted for Bonferroni correction.

Results

Heavier smoking was associated with higher risk of CRC overall and stratified by individual markers (Ptrend < .001). The associations differed statistically significantly between all molecular subtypes, which was the most statistically significant for CIMP and BRAF. Compared with never-smokers, smokers in the fourth quartile of pack-years had a 90% higher risk of CIMP-positive CRC (odds ratio = 1.90, 95% confidence interval = 1.60 to 2.26) but only 35% higher risk for CIMP-negative CRC (odds ratio = 1.35, 95% confidence interval = 1.22 to 1.49; Pdifference = 2.1 x 10-6). The association was also stronger in tumors that were CIMP positive, MSI high, or KRAS wild type when combined (Pdifference < .001).

Conclusion

Smoking was associated with differential risk of CRC subtypes defined by molecular characteristics. Heavier smokers had particularly higher risk of CRC subtypes that were CIMP positive and MSI high in combination, suggesting that smoking may be involved in the development of colorectal tumors via the serrated pathway.
Experimental study on two consecutive droplets impacting onto an inclined solid surface
Authors: Chen Chun-Kuei , Chen Sheng-Qi , Yan Wei-Mon , Li Wen-Ken , Lin Ta-Hui
Published: June 2021

The present study is concerned with the experimental impingement of two consecutive droplets on an inclined solid surface. Attention is mainly paid to the effects of impingement timing with various oblique angles (Φ) of the surface on the impact phenomena, which mainly affect the maximum droplet spreading diameter. The investigation considers four impingement scenarios differentiated by impingement timing, namely Case 1: single-droplet impingement; Case 2 of Δt1: the moment when the leading droplet starts spreading along the oblique surface; Case 3 of Δt2: the moment when the leading droplet reaches its maximum spreading; and Case 4 of Δt3: the moment when the leading droplet starts retracting. It is observed that deformation behavior of two successive droplets impacting on the inclined surface experiences a complex asymmetric morphology evolution due to the enhancement of gravity effect and various conditions of the impingement timing. The merged droplet becomes slender with increasing oblique surface angle in the final steady shape, causing the decrease in the value of front and back contact angles. The impingement timing has a significant influence on the change of the maximum height of the merged droplet. The coalesced droplet spreads to the maximum dimensionless width diameter at Δt = Δt2 and the oblique angle of Φ = 45°, but reaches the maximum dimensionless height for Δt = Δt2 at Φ = 30°. The front contact angles converge to a fixed value eventually for all conditions of impingement timing, and the values become lower with the increasing surface inclination.
A fast semidiscrete optimal transport algorithm for a unique reconstruction of the early Universe
Authors: Levy Bruno , Mohayaee Roya , von Hausegger Sebastian
Published: June 2021

We leverage powerful mathematical tools stemming from optimal transport theory and transform them into an efficient algorithm to reconstruct the fluctuations of the primordial density field, built on solving the Monge-Ampère-Kantorovich equation. Our algorithm computes the optimal transport between an initial uniform continuous density field, partitioned into Laguerre cells, and a final input set of discrete point masses, linking the early to the late Universe. While existing early universe reconstruction algorithms based on fully discrete combinatorial methods are limited to a few hundred thousand points, our algorithm scales up well beyond this limit, since it takes the form of a well-posed smooth convex optimization problem, solved using a Newton method. We run our algorithm on cosmological N-body simulations, from the AbacusCosmos suite, and reconstruct the initial positions of $\mathcal {O}(10^7)$ particles within a few hours with an off-the-shelf personal computer. We show that our method allows a unique, fast, and precise recovery of subtle features of the initial power spectrum, such as the baryonic acoustic oscillations.
Recombination machinery engineering facilitates metabolic engineering of the industrial yeast Pichia pastoris
Authors: Cai Peng , Duan Xingpeng , Wu Xiaoyan , Gao Linhui , Ye Min , Zhou Yongjin J
Published: June 2021

The industrial yeast Pichia pastoris has been harnessed extensively for production of proteins, and it is attracting attention as a chassis cell factory for production of chemicals. However, the lack of synthetic biology tools makes it challenging in rewiring P. pastoris metabolism. We here extensively engineered the recombination machinery by establishing a CRISPR-Cas9 based genome editing platform, which improved the homologous recombination (HR) efficiency by more than 54 times, in particular, enhanced the simultaneously assembly of multiple fragments by 13.5 times. We also found that the key HR-relating gene RAD52 of P. pastoris was largely repressed in compared to that of Saccharomyces cerevisiae. This gene editing system enabled efficient seamless gene disruption, genome integration and multiple gene assembly with positive rates of 68–90%. With this efficient genome editing platform, we characterized 46 potential genome integration sites and 18 promoters at different growth conditions. This library of neutral sites and promoters enabled two-factorial regulation of gene expression and metabolic pathways and resulted in a 30-fold range of fatty alcohol production (12.6–380 mg/l). The expanding genetic toolbox will facilitate extensive rewiring of P. pastoris for chemical production, and also shed light on engineering of other non-conventional yeasts.
The cooperative assembly of shelterin bridge provides a kinetic gateway that controls telomere length homeostasis
Authors: Liu Jinqiang , Hu Xichan , Bao Kehan , Kim Jin-Kwang , Zhang Catherine , Jia Songtao , Qiao Feng
Published: June 2021

Shelterin is a six-protein complex that coats chromosome ends to ensure their proper protection and maintenance. Similar to the human shelterin, fission yeast shelterin is composed of telomeric double- and single-stranded DNA-binding proteins, Taz1 and Pot1, respectively, bridged by Rap1, Poz1 and Tpz1. The assembly of the proteinaceous Tpz1-Poz1-Rap1 complex occurs cooperatively and disruption of this shelterin bridge leads to unregulated telomere elongation. However, how this biophysical property of bridge assembly is integrated into shelterin function is not known. Here, utilizing synthetic bridges with a range of binding properties, we find that synthetic shelterin bridge lacking cooperativity requires a linker pair that matches the native bridge in complex lifespan but has dramatically higher affinity. We find that cooperative assembly confers kinetic properties on the shelterin bridge allowing disassembly to function as a molecular timer, regulating the duration of the telomere open state, and consequently telomere lengthening to achieve a defined species-specific length range.
The ‘Ethical’ COVID-19 Vaccine is the One that Preserves Lives: Religious and Moral Beliefs on the COVID-19 Vaccine
Authors: Giubilini Alberto , Minerva Francesca , Schuklenk Udo , Savulescu Julian
Published: June 2021

Although the COVID-19 pandemic is a serious public health and economic emergency, and although effective vaccines are the best weapon we have against it, there are groups and individuals who oppose certain kinds of vaccines because of personal moral or religious reasons. The most widely discussed case has been that of certain religious groups that oppose research on COVID-19 vaccines that use cell lines linked to abortions and that object to receiving those vaccine because of their moral opposition to abortion. However, moral opposition to COVID-19 vaccine research can be based on other considerations, both secular and religious. We argue that religious or personal moral objections to vaccine research are unethical and irresponsible, and in an important sense often irrational. They are unethical because of the risk of causing serious harm to other people for no valid reason; irresponsible because they run counter to individual and collective responsibilities to contribute to important public health goals; and in the case of certain kinds of religious opposition, they might be irrational because they are internally inconsistent. All in all, our argument translates into the rather uncontroversial claim that we should prioritize people’s lives over religious freedom in vaccine research and vaccination roll out.
HBI transcription factor-mediated ROS homeostasis regulates nitrate signal transduction
Authors: Chu Xiaoqian , Wang Jia-Gang , Li Mingzhe , Zhang Shujuan , Gao Yangyang , Fan Min , Han Chao , Xiang Fengning , Li Genying , Wang Yong , Yu Xiang , Xiang Cheng-Bin , Bai Ming-Yi
Published: June 2021

Nitrate is both an important nutrient and a critical signaling molecule that regulates plant metabolism, growth, and development. Although several components of the nitrate signaling pathway have been identified, the molecular mechanism of nitrate signaling remains unclear. Here, we showed that the growth-related transcription factors HOMOLOG OF BRASSINOSTEROID ENHANCED EXPRESSION2 INTERACTING WITH IBH1 (HBI1) and its three closest homologs (HBIs) positively regulate nitrate signaling in Arabidopsis thaliana. HBI1 is rapidly induced by nitrate through NLP6 and NLP7, which are master regulators of nitrate signaling. Mutations in HBIs result in the reduced effects of nitrate on plant growth and ∼22% nitrate-responsive genes no longer to be regulated by nitrate. HBIs increase the expression levels of a set of antioxidant genes to reduce the accumulation of reactive oxygen species (ROS) in plants. Nitrate treatment induces the nuclear localization of NLP7, whereas such promoting effects of nitrate are significantly impaired in the hbi-q and cat2 cat3 mutants, which accumulate high levels of H2O2. These results demonstrate that HBI-mediated ROS homeostasis regulates nitrate signal transduction through modulating the nucleocytoplasmic shuttling of NLP7. Overall, our findings reveal that nitrate treatment reduces the accumulation of H2O2, and H2O2 inhibits nitrate signaling, thereby forming a feedback regulatory loop to regulate plant growth and development.
Towards genetically encoded sensors for nitric oxide bioimaging in planta
Authors: Safavi-Rizi Vajiheh
Published: June 2021

Towards genetically encoded sensors for nitric oxide bioimaging in planta
The Conundrum of Kappa and Why Some Musculoskeletal Tests Appear Unreliable Despite High Agreement: A Comparison of Cohen Kappa and Gwet AC to Assess Observer Agreement When Using Nominal and Ordinal Data
Authors: Cibulka Michael T , Strube Michael J
Published: June 2021

In clinical practice, physical therapists often use different kinds of tests and measures in the assessment of their patients. For therapists to have confidence when using their tests and measures, an important attribute is having intratester and intertester reliability. Studies that assess reliability are cases of observer agreement. Many studies have been performed assessing observer agreement in the physical therapy literature. The most commonly used method to assess observer agreement studies that use nominal or ordinal data is the statistical method suggested by Cohen and the corresponding reliability coefficient, Cohen kappa. Recently, Cohen kappa has undergone scrutiny because of what is called kappa paradox, which occurs when observer agreement is high but the resulting kappa value is low. Another paradox also occurs when asymmetries exist between raters on their disagreements, resulting in a higher kappa value. In the physical therapy literature, there are numerous examples of this problem, which can often lead to misunderstanding the meaning of the data. This Perspective examines how and why these problems occur and suggests an alternative method for assessing observer agreement.
Business Restrictions and COVID-19 Fatalities
Authors: Spiegel Matthew , Tookes Heather
Published: June 2021

We collect a time-series database of business and related restrictions for every county in the United States from March through December 2020. We find strong evidence consistent with the idea that employee mask policies, mask mandates for the general population, restaurant and bar closures, gym closures, and high-risk business closures reduce future fatality growth. Other business restrictions, such as second-round closures of low- to medium-risk businesses and personal care/spa services, did not generate consistent evidence of lowered fatality growth and may have been counterproductive.
A New Pipeline for Removing Paralogs in Target Enrichment Data
Authors: Zhou Wenbin , Soghigian John , Xiang Qiu-Yun (Jenny)
Published: June 2021

Target enrichment (such as Hyb-Seq) is a well-established high throughput sequencing method that has been increasingly used for phylogenomic studies. Unfortunately, current widely used pipelines for analysis of target enrichment data do not have a vigorous procedure to remove paralogs in target enrichment data. In this study, we develop a pipeline we call Putative Paralogs Detection (PPD) to better address putative paralogs from enrichment data. The new pipeline is an add-on to the existing HybPiper pipeline, and the entire pipeline applies criteria in both sequence similarity and heterozygous sites at each locus in the identification of paralogs. Users may adjust the thresholds of sequence identity and heterozygous sites to identify and remove paralogs according to the level of phylogenetic divergence of their group of interest. The new pipeline also removes highly polymorphic sites attributed to errors in sequence assembly and gappy regions in the alignment. We demonstrated the value of the new pipeline using empirical data generated from Hyb-Seq and the Angiosperms353 kit for two woody genera Castanea (Fagaceae, Fagales) and Hamamelis (Hamamelidaceae, Saxifragales). Comparisons of data sets showed that the PPD identified many more putative paralogs than the popular method HybPiper. Comparisons of tree topologies and divergence times showed evident differences between data from HybPiper and data from our new PPD pipeline. We further evaluated the accuracy and error rates of PPD by BLAST mapping of putative paralogous and orthologous sequences to a reference genome sequence of Castanea mollissima. Compared to HybPiper alone, PPD identified substantially more paralogous gene sequences that mapped to multiple regions of the reference genome (31 genes for PPD compared with 4 genes for HybPiper alone). In conjunction with HybPiper, paralogous genes identified by both pipelines can be removed resulting in the construction of more robust orthologous gene data sets for phylogenomic and divergence time analyses. Our study demonstrates the value of Hyb-Seq with data derived from the Angiosperms353 probe set for elucidating species relationships within a genus, and argues for the importance of additional steps to filter paralogous genes and poorly aligned regions (e.g., as occur through assembly errors), such as our new PPD pipeline described in this study. [Angiosperms353; Castanea; divergence time; Hamamelis; Hyb-Seq, paralogs, phylogenomics.]
Titratable residues in the extracellular domain of human gamma‐ENaC contribute to H+ regulation of channel activity
Authors: Collier Daniel M , Snyder Peter M
Published: June 2021

The extracellular domain of the epithelial sodium channel (ENaC) plays a critical role in tuning channel activity in response to dynamic extracellular conditions. We have previously demonstrated that H+ have dual effects on ENaC activity. H+ increase ENaC activity by decreasing Na+ self‐inhibition. H+ also inhibit ENaC activity by increasing Cl− inhibition. In the current work, we investigated the mechanisms by which H+ stimulate ENaC. We took advantage of our previous finding that rat ENaC has a reduced response to pH compared to human ENaC. By expressing combinations of human and rat subunits, we localized the difference to the γ subunit. Our strategy was to mutate H+ titratable residues in the extracellular domain of human γENaC that are not conserved in rat γENaC. We expressed the mutant γENaC cDNAs (with wild type α‐ and βENaC) in Xenopus oocytes and tested the effect of pH changes on amiloride‐sensitive Na+ current (by TEVC at −60 mV). We identified a group of residues in the extracellular domain of γENaC (D164, Q165, D166, E292, E315, E335, H439, and E455) that, when individually mutated to Ala, decreased H+ activation of ENaC. Furthermore, combining single mutations resulted in an additive decrease in the pH response. The data demonstrate that multiple residues in human γENaC are required for regulation by pH.
Identification of pH‐sensitive currents in Drosophila sensory neurons
Authors: Boiko Nina , Eaton Benjamin A. , Stockand James D.
Published: June 2021

Acid‐sensing ion channels (ASIC) are proton‐gated Na+ channels that belong to the epithelial Na+ channel/degenerin (ENaC/Deg) superfamily. ASICs contribute to mechanosensation, nociception and synaptic plasticity. Pickpocket, PPK, are ENaC/Deg homologs in Drosophila neurons. We ask here whether PPK function as acid‐sensing channels with a view towards identify physiological function. Using whole cell patch‐clamping, we identified two acid‐sensing Na+ currents in multiple dendritic (md) neurons of Drosophila melanogaster. One current is robust and activated by pronounced acidification. Amiloride, a nonspecific antagonist of ENaC/Deg channels, reduces this pH‐sensitive current in a dose‐dependent manner. In addition, a distinct transient Na+ current (Na+tr) is present in md neurons. This current is activated by relief from inhibition subsequent to removal of amiloride following extracellular acidification. The standing acid‐sensing current was retained in md neurons of PPK−/− flies. In contrast, Na+tr was absent in md neurons of PPK−/− flies implying that PPK is part of the pH‐sensitive channel complex underlying this transient current. Phenotype assay of PPK−/− flies is clarifying the physiological role of PPK: preliminary results suggest a role in control of courtship behavior.
Paracrine erythropoietin (EPO) signaling and anti‐apoptosis in the lungs of guinea pigs exposed to high altitude (HA)
Authors: Hsia Connie C.W. , Zhang Qiuyang Daniel , Foster David J. , Ravikumar Priya , Powell Frank L.
Published: June 2021

HA residence stimulates erythropoiesis as well as lung growth and remodeling in young animals, suggesting parallel upregulation of endocrine and paracrine/autocrine EPO signaling. To examine this possibility, we exposed weanling male guinea pigs to 3,800 m HA for 3 d, 7 d or 1 mo compared to matched controls at sea level (SL) (n = 5–8 each), and measured mRNA and/or protein expression of hypoxia‐inducible factors (HIF‐1alpha and ‐2alpha), EPO and its receptor (EPOR), Jak2, VEGF and its receptors (−R1, −R2), endothelin‐1 (ET‐1), proliferation marker (PCNA), and an apoptotic suppressor (Bcl‐2) and a promoter (BAD). Compared to SL, HA‐exposed lungs showed 100% and 50% higher HIF‐1alpha mRNA at 3 and 7 d and 55% lower HIF‐2alpha mRNA at 3 d, although HIF‐2alpha protein was 120% higher. Lung EPO mRNA and cytoplasmic EPOR protein increased 150% and 25% at 3 d without significant changes in membrane EPOR or Jak2. Plasma and urine EPO protein increased 56% and 160% at 3 d. Lung VEGF‐R2 protein increased 72% at 3 d while VEGF increased 27% at 7 d. ET‐1 mRNA increased 3‐fold at 3 and 7 d. BAD was 50% lower and Bcl‐2 100% higher at 3 d. PCNA was unaltered. All changes resolved by 1 mo. Thus, short‐term HA exposure stimulates both endocrine and paracrine/autocrine EPO signaling. Enhanced EPO signaling in the lung occurs primarily at the ligand but not the receptor level and is associated with anti‐apoptosis. Support: NIH R01 HL045716.
A pathway impact analysis approach to identify context‐specific signaling networks consistent with differential gene expression data
Authors: Vadigepalli Rajanikanth
Published: June 2021

The objective of the present study is to integrate the information on signaling network structures with differential gene expression data sets to predict functional changes in the cellular pathways that are operational under those conditions. We developed a novel approach that starts with a user‐specified pathway structure, generates thousands of alternative pathway structures and evaluates each network for consistency with the gene expression data based on the statistical significance of a perturbation impact factor metric. Our approach removes the interactions in the initial network that are not consistent with the gene expression changes, and yields context‐specific signaling pathways. We benchmarked our approach using biologically motivated synthetic networks as well as a Gq‐protein coupled receptor pathway. Our results indicate that (1) networks with high consistency are likely to arise by differential expression of activators and inhibitors in opposing directions, and (2) nodes with upstream activation and inhibitory interactions significantly affect the network consistency. Research Support: NIH ‐ R01 GM083108 and R33 HL088283.
A novel approach to identifying the major disease processes associated with diabetic nephropathy
Authors: McMahon Kevin Wyatt , Perez Jerry , Prabhakar Sharma
Published: June 2021

Most complex diseases can be summarized by a small number of “disease processes.” Identifying those processes can lead to improved understanding and therapeutic options. We have devised a new procedure for disease process discovery using diabetic nephropathy (DN) as the example. Our method consists of three steps: 1) an effect size is calculated for the disease of interest and all other publicly‐available Affymetrix gene array experimental conditions (using a condor grid system), 2) a novel distance statistic is calculated for each gene to identify genes that are similarly affected in the experimental condition of interest and DN 3)the counts of the gene ontology (GO) terms associated with these similarly affected genes serve as the basis for class discovery (using positioning around medioids) followed by variable selection using a Wilcox test ranking procedure. We divide DN into 3 different disease processes (cytoskeltal/morphological changes, inflammation, and response to hyperglycemia). Important GO terms in distinguishing between these 3 factors included actin filament bundle assembly, defense response, and response to nutrient. Since these disease processes are well known components of the pathology of diabetic nephropathy, this suggests we have developed a powerful method for identifying the major components of complex disease.
Force closure of the sacroiliac joint: cadaver study of the extent of gluteus maximus attachment on the sacrum and relative orientation of muscle fibers
Authors: Seim Sophia , Carey Fred , Andrews Ron , Jones Beth Moody
Published: June 2021

The lumbosacral region has many muscles that may stabilize the region during functional activities via force closure (FC). The sacroiliac joint (SIJ) is a key element of this region. The role of individual muscles in FC of the SIJ is poorly understood. We undertook an anatomical study to examine the contribution of the gluteus maximus (GM) to FC.

The posterior superior iliac spine (PSIS), sacrum, coccyx, and muscle fiber attachments of the GM were exposed on 30 embalmed cadavers. Sacral dimensions and specific GM muscle fiber attachment sites and their length were recorded. The angles of GM fibers relative to the SIJ were measured by goniometric technique at three specific points on the sacrum. All measurements were taken relative to a reference line connecting the most prominent point of the PSIS to the coccyx (line A).

The mean width of GM attachment onto the sacrum at line A was 11.1 cm. At the midpoint of line A, GM extends past the SIJ attaching to the sacrum with a mean length of 3.13 cm and a mean angle of fiber alignment of 90 degrees. The orientation of GM fibers measured at the PSIS was 85.4 degrees.

The GM's considerable length of attachment onto the sacrum and its angles of fiber alignment suggest that it may play a significant role in stabilization and force closure of the SIJ. Inclusion of GM recruitment and strengthening exercises may be beneficial for patients diagnosed with sacroiliac dysfunction.

Grant Funding Source: none
Anatomical Variation in the Maximum Bending Moment of Metacarpals 2 through 5
Authors: Adar Tony , Baral Sumit , Hayes Westley , Saha Subrata , Pagano Anthony , Marquez Samuel
Published: June 2021

Metacarpals (MC) two through five function as transmitters of force in activities involving the hand, making them susceptible to various types of fractures. An important clinical relevance is that the prevalence of metacarpal fractures has been estimated to be about 39% of all fractures of the hand. This study explores the relationship between maximum bending moment (MBM) and mid‐shaft diameter of embalmed cadaveric MC two through five in order to evaluate structural strength and fracture risk. Twelve donor bodies were examined followed by disarticulation and removal of all soft tissue of MC 2–5. All specimens were radiographed to assess for preexisting fractures and to rule out osteopenic changes. The samples were mechanically tested in three point bending using a Instron 8874 (Norwood, MA) Servohydraulic Test System. MBM was calculated for each MC and they ranged from 6.81 N·m to 3.60 N·m of MC2 to MC5 respectively. Our preliminary findings revealed a high positive correlation between mid‐shaft diameter and MBM (r2 = 0.96; p<0.05) suggesting that bending stresses resulting in MC fractures may be predicted with evaluation of mid‐shaft diameter, which may be of clinical importance.

Grant Funding Source: SUNY Downstae College of Medicine
A suitable model to investigate the regeneration of transplanted hepatocytes
Authors: Itagaki Hiroko , Kitahara Shuji , Ezaki Taichi
Published: June 2021

Aim

To seek for a suitable site for isolated hepatocytes to proliferate with a high reliability, we established a mouse transplantation model and morphologically characterized the regeneration of transplanted ectopic hepatocytes.

Materials & Methods

Adult male C57BL/6 mice were used as hepatocyte donors and hepatocytes were transplanted into mesenteric fat pads of C57BL/6‐Tg (CAG‐GFP) mice as recipients. At various times after transplantation of isolated hepatocytes (1.5×107 cells per site), mice were sacrificed and cryosections were made. Multiple‐immunostaining of various markers was employed for the 3‐dimentional imaging by the confocal laser‐scanning microscopy.

Results & Discussion

Three weeks after transplantation, the transplanted hepatocytes vigorously regenerated to cell masses raging from 350μm to 2500μm in diameter. They are also acoompanied by the development of hepatic sinusoids of donor origin. The endotherial and type‐IV collagen linings of the sinusoids were comparable to those of the normal liver. Comparing with other transplantation sites, this model, therefore, provides a useful tool to investigate the regeneration of the liver structure and function after transplantation.
Convergence of visceral and somatic sensory information in the sacral cord: morphologic evidence for referred pain
Authors: Li Yun‐Qing , Zhang Ming‐Ming , Kou Zhen‐Zhen
Published: June 2021

The somatic foci of pelvic organs referred pain are often and mainly distributed along the anteriolateral aspect of the lower limbs. Our study was designed to investigate the neural pathways and their transmitters for the pelvic visceral referred pain. By using tracing techniques, we observed that neurons in the sacral dorsal commissural nucleus (SDCN) innervated by primary afferents originated from both pelvic nerve and sciatic nerve and projected to parabrachial nucleus (PBN). Excitatory postsynaptic potentials (EPSPs) were elicited by the electrical stimulation of the pelvic or sciatic nerves in the SDCN neurons tested by patch clamp recording. After visceral noxious stimulation, there were more Fos‐immunopositive (Fos‐ip) neurons in the SDCN than that in the dorsal horn. SPR‐ip neurons with FOS‐ip nuclei projected to the PBN and contacted with SP‐ip terminals. Under electron microscope, we found that SP‐ip terminals or ENK‐, GABA‐, Gly‐ip terminals to make synapses with SDCN neurons projected to the PBN. These results indicated that: (1) SDCN might be the convergence point of the referred pain associated with pelvic visceral organs to lower limbs; (2) The projection from the SDCN to the PBN might contribute to the pelvic visceral information transmission; (3) SP, ENK, GABA and Gly are important for the pelvic visceral information transmission and regulation.

National Natural Science Foundation of China
An immunohistochemical study on the arrangement of sympathetic fibers within the human common fibular nerve
Authors: Tompkins Rebecca , Melling Jamie , Shoemaker J Kevin , Wilson Timothy D
Published: June 2021

Physiologists often use muscle sympathetic neural activity (MSNA) in the common fibular nerve (CFN) to measure sympathetic drive within lower limb peripheral nerves. It is understood that significant alterations in MSNA activity occur with age, diabetes, and vascular disease. Little is known, however, about sympathetic fiber distribution in the lower limb nerves of humans. In order to describe the general sympathetic arrangement in the CFN, immunohistochemical techniques were used on 5μm transverse sections of formaldehyde‐fixed CFN taken from human cadavers within 24 hours of death. By staining for tyrosine hydroxylase the arrangement and number of sympathetic fibers can be determined. From microneurographic records it is hypothesized that the sympathetic fibers are arranged in randomly dispersed bundles, each of approximately 10–40 fibers. Results from this study will quantify autonomic fiber distribution in the CFN. This will better enable physiologists to relate autonomic neuroanatomy to microneurographic outcomes in the CFN.

Grant Funding Source: Departmental
The augmentative action of taurine on the differentiation of C2C12 cells to myotube
Authors: Miyazaki Teruo , Honda Akira , Ikegami Tadashi , Shirai Mutsumi , Miyakawa Shumpei , Matsuzaki Yasushi
Published: June 2021

Background/Aim

Incomplete muscular development is induced by deficiency of taurine that is endogenously and abundantly supplied thought placenta and milk via a specific transporter (TAUT). This study investigated the influence of taurine on the differentiation of myoblast.

Method

Confluent mice myoblast (C2C12) cell was exposed to up to 20mM taurine with a differentiation medium for up to a week. The differentiation markers were evaluated by RT‐qPCR, fluorescence immunohistochemical stain or Western blot. The influences of cellular taurine uptake inhibition on the myoblast differentiation were also examined.

Result

The fusion index and maximal diameter in the MHC‐positive myotubes were significantly increased, in particular 6‐fold in 20mM taurine, compared to those in 0mM taurine. The phosphorylations of Akt and p38MAPK were increased and decreased, respectively, by taurine. Furthermore, the expression of myostatin protein was suppressed by taurine exposure. The differentiations in exposing to 20mM taurine were significantly inhibited by taut siRNA or β‐alanine; a taurine transport competitor.

Conclusion

Exogenous taurine might play a key role on the skeletal muscle differentiation during development period. The effects of taurine should contribute the recovery from muscle damages and differentiation of satellite cells.
Novel Insight into the Staining and Counterstaining Properties of Lawsonia inermis (Henna) in Histological Preparations
Authors: Alawa Judith Nkiruka , Aribiyun Yomi S , Adetiba Bamidele , Alawa Clement B.I.
Published: June 2021

Lawsonia inermis commonly called Henna is a naphthoquinone‐rich plant which is exploited as a natural dye for hair colouring and cosmetic purposes for nails and skin. Histological staining properties of Lawsonia inermis was studied using Wistar rat specimens of skin, liver, brain, kidney and intestine fixed in neutral buffered formalin and prepared for light microscopy. Stain was prepared as 0.5% extract of leaves of Lawsonia inermis dissolved in 5% aqueous alcohol and mordanted with potassium alum (aluminium potassium sulphate). The tested extract stained cell cytoplasm brown and gave consistent reproducible results. Nuclei did not pick up the stain suggesting that it is an acidic stain similar to eosin. Stain showed great affinity for keratin and muscle cells which stained deeply. Lawsonia inermis when used as a counterstain to haematoxylin showed marked contrast and clarity which was comparable to routine H&E preparations and was heat stable. This indicates that Lawsonia inermis could be developed as an acidic stain or a counterstain to haematoxylin in histological techniques for highlighting structures in biological tissues.
Clathrin‐associated Endocytic Proteins Eps15 and Epsin are Crucial for the Formation of Enteropathogenic E. coli (EPEC) pedestals
Authors: Lin Ann E. , Benmerah Alexandre , Guttman Julian A.
Published: June 2021

Enteropathogenic E.coli (EPEC) are primarily extracellular pathogens that cause severe infantile diarrhea. The hallmark of EPEC pathogenesis is the generation of actin‐rich structures known as pedestals. Surprising evidence has revealed that while they persistently reside at an extracellular space, EPEC require the receptor‐mediated endocytosis protein, clathrin, for pedestal formation. In order to understand the strategies EPEC utilize to usurp endocytosis, we examined the roles of three major endocytic components: AP‐2, Eps15 and epsin, during pedestals formation. We found that pedestal formations required the recruitment of Eps15 and epsin, but not AP‐2. AP‐2 is known to provide linkage between Eps15, epsin and clathrin during the clathrin‐coated pit assembly process to complete endocytosis. Together, these results reveal a novel endocytic protein manipulation strategy employed by EPEC, supporting the recent paradigm that endocytic proteins are important for EPEC‐mediated disease.

Grant Funding Source: CIHR
Genome‐wide screen identifies genes required for Francisella invasion in non‐phagocytic cells
Authors: Lo Karen Y , Nano Francis E , Guttman Julian A
Published: June 2021

The intracellular bacterium Francisella tularensis subspecies tularensis (F. tularensis) is the causative agent of tularemia, a disease that can result in 30–35% mortality if left untreated. Due to F. tularensis' high infectivity and lethality the Centers for Disease Control and Prevention has classified it as a Biosafety Level 3 pathogen and a Category A Select agent. F. tularensis host cell invasion is crucial for their pathogenesis as without invasion disease does not occur. Upon entry into their host, the bacteria colonize both macrophages and epithelial cells. Despite significant research advances in macrophage infections, the study of epithelial infections has lagged behind. To investigate F. tularensis genes involved in epithelial cell invasion, we utilized a biosafety level 2 bacterial surrogate called F. tularensis subspecies novicida (F. novicida). A F. novicida transposon mutant library was screened to assess their invasion in hepatocytes. We selected for mutants with reduced intracellular growth using gentamycin‐based invasion/survival assays and confirmed our results microscopically using a subset of mutants. We discovered 232 mutants had significantly reduced invasion/survival levels as compared to wild type F. novicida infections. Ultimately our findings open a new door to the discovery of novel targets for the development of therapeutics and prophylactics.

Grant Funding Source: CIHR and NSERC
α‐II‐spectrin as a marker for the striated organelle in inner ear hair cells
Authors: Chidavaenzi Robstein L , Vranceanu Florin , Lysakowski Anna
Published: June 2021

Located in the subcuticular zone of vestibular and auditory hair cells, the striated organelle's form, occurrence and function remain largely unknown. Studying its architecture by employing serial section transmission electron microscopy, intermediate voltage electron microscopy and electron microscope tomography, we have come to recognize that the striated organelle (SO) is a cytoskeletal apparatus shaped like an inverted cone that is made up of alternating thin and thick filaments. It is associated with microtubules, endoplasmic reticular and large mitochondria that have polarized, lamellar cristae. It is attached to both stereociliar rootlets and the cell membrane, raising the prospect of an active role in hair cell mechanotransduction. As we attempt to answer questions pertaining to its function, unraveling its protein composition is essential. In this study we examined the development of α‐II‐spectrin immunoreactivity in rats (aged P1 to adult). Our confocal data show cuticular plate labeling in both type I and II hair cells in the cristas, utricules and saccules. More telling is the localization of α‐II‐spectrin in the region where the SO occurs. The staining pattern here is less intense in younger animals and more pronounced in the adult.

Grant Funding Source: NIH R01‐DC02521
Regulation of pyruvate dehydrogenase kinase 4 (PDK4) by CCAAT/enhancer‐binding protein beta (C/EBPb)
Authors: SHARMA PRAGYA , ATTIA RAMY R , SONG SHULAN , ELAM MARSHALL B , COOK GEORGE A , JANSSEN RACHEL C , FRIEDMAN JACOB E , PARK EDWARDS A
Published: June 2021

Thyroid hormone (T3) regulates various aspects of hepatic metabolism. C/EBPβ modulates the expression of multiple hepatic genes including those involved in hepatic gluconeogenesis. The conversion of pyruvate to acetyl‐CoA in mitochondria is catalyzed by the pyruvate dehydrogenase complex (PDC). Activity of PDC is decreased via phosphorylation by the pyruvate dehydrogenase kinases (PDK). Here we investigated the role of the CCAAT/enhancer‐binding protein β (C/EBPβ) in the T3 regulation of PDK4 gene expression. Our data indicate that C/EBPβ induces PDK4 gene expression and decreases PDC activity. This transcriptional induction is mediated through C/EBPβ binding sites in the PDK4 promoter. T3 increased C/EBPβ abundance in primary rat hepatocytes. Knockdown of C/EBPβ with siRNA diminished the T3 induction of the PDK4. Our data suggest that C/EBPβ is an important accessory factor for the T3 activation of PDK4 gene.
The Na/K‐ATPase/Caveolin‐1 Interaction Regulates Cell Growth Via A Cholesterol‐Dependent Mechanism
Authors: Dong Shuai , Chen Yiliang , Xie Zijian
Published: June 2021

The Na/K‐ATPase α1 subunit contains a highly conserved caveolin‐1 binding motif (CBM). We have previously shown that the interaction between the α1 CBM and caveolin‐1 regulates the formation of caveolae, and intracellular trafficking and distribution of cholesterol. Here we demonstrate an important role of this interaction in control of cell growth. First, we find that knockdown of either Na/K‐ATPase or caveolin‐1 inhibits cell proliferation and that this inhibition is greatly enhanced by modest depletion of cellular cholesterol. Second, the expression of wild‐type α1, but not the CBM mutant, is capable of restoring normal growth in LLC‐PK1 cells. Finally, we find that the expression of N‐terminus of the α1 subunit inhibits the interaction between the Na/K‐ATPase and caveolin‐1. Consequently, it alters normal trafficking of caveolin‐1 and cholesterol, and inhibits cell growth. Taken together, our new findings demonstrate that the highly conserved caveolin binding motif of the Na/K‐ATPase plays an important role in regulation of cell growth by affecting the trafficking and distribution of caveolin‐1 and cholesterol (supported by NIH grants HL36573 and GM78565).
Effects of LPA antagonist and PLD inhibitor on proliferation of human prostate cancer cell lines
Authors: Zhang Zhihong , Meier Kathryn E
Published: June 2021

Lysophosphatidic acid (LPA) is a phospholipid mediator that binds to G protein‐coupled receptors. LPA receptors can activate phospholipase D (PLD), and PLD can participate in LPA production. Previous work from our lab has characterized LPA receptors and response in human prostate cancer cell lines, showing that LPA and EGF can activate PLD and induce LPA production. The current study addresses whether LPA antagonists and/or PLD inhibitors interfere with prostate cancer cell growth. Three human prostate cancer cell lines were used: PC‐3, DU145, and LNCaP. Proliferation assays showed that serum, LPA, and EGF stimulate proliferation of PC‐3 and DU145 cells. Only serum and EGF stimulate proliferation of LNCaP, consistent with published data showing that LNCaP does not respond to LPA. The effects of an LPA receptor antagonist (Ki16425) and a PLD inhibitor (FIPI) were tested. Ki16425 significantly inhibits proliferation of PC‐3 and DU145 in response to LPA and EGF. FIPI inhibits LPA‐ and EGF‐induced growth in PC‐3 and DU145, and also inhibits EGF‐induced proliferation of LNCaP. The observation that an LPA antagonist inhibits EGF response suggests that cross‐talk between EGF and LPA receptors is functionally important. Moreover, the finding that a PLD inhibitor can interfere with proliferation suggests that PLD is a potential therapeutic target in prostate cancer. Supported by an ITHS/INBRE Translational Seed Grant.