bluuebunny/crossref_metadata_2025 huggingface dataset:
Trans-epithelial Fluid Pumping Performance of Renal Epithelial Cells and Mechanics of Cystic Expansion
Authors: Choudhury Mohammad Ikbal , Li Yizeng , Mistriotis Panagiotis , Vasconcelos Ana , Dixson Eryn , Yang Jing , Benson Morgan , Maity Debonil , Walker Rebecca , Martin Leigha , Koroma Fatima , Qian Feng , Konstantopoulos Konstantinos , Woodward Owen , Sun Sean
Published: May 2021

Using a novel microfluidic platform to recapitulate fluid transport activity of kidney cells, we report that renal epithelial cells can actively generate hydraulic pressure gradients across the epithelium. The fluidic flux declines with increasing hydraulic pressure until a stall pressure, at which the flux vanishes--in a manner similar to mechanical fluidic pumps. The developed pressure gradient translates to a force of 50-100 nanoNewtons per cell. For normal human kidney cells, the fluidic flux is from apical to basal, and the pressure is higher on the basal side. For human Autosomal Dominant Polycystic Kidney Disease (ADPKD) cells, the fluidic flux is reversed from basal to apical with a higher stall pressure. Molecular studies and proteomic analysis reveal that renal epithelial cells are sensitive to hydraulic pressure gradients, developing different expression profiles and spatial arrangements of ion exchangers and the cytoskeleton in different pressure conditions. These results, together with data from osmotic and pharmacological perturbations of fluidic pumping, implicate mechanical force and hydraulic pressure as important variables during morphological changes in epithelial tubules, and provide further insights into pathophysiological mechanisms underlying the development of high luminal pressure within renal cysts.
Cinnamamide-Chalcone Derivatives as CDK2 Inhibitors: Synthesis, Pharmacological Evaluation, and Molecular Modelling Study
Authors: Joshi Akshada J. , Bhojwani Heena R. , Joshi Urmila , Begwani Khushboo V. , Wagal Ojas S. , Sathaye Sadhana S. , Kanchan Divya M.
Published: May 2021

A series of 13 novel cinnamamide-chalcone derivatives (2a-2m) were synthesized and evaluated for their antiproliferative activity against MCF-7, K562, U373MG, and HT-29 cell lines by SRB assay. Considering the activities on MCF-7 cell line, eight compounds were tested for the in-vitro CDK2 inhibition and four (2g, 2h, 2k and 2l) were found to possess good activity (IC50<10µM). These four compounds were tested on EGFR kinase to assess the selectivity towards CDK2 and were found be nearly two times more selective. To corroborate the in-vitro enzyme assay data with binding, the compounds were docked into the CDK2 and EGFR using Glide software. The docking studies reveal that all eight compounds form hydrogen bonds with Lys33 (β-3 region) and Leu83 (hinge region) in CDK2 and the docking scores correlate well with the IC50 values. The most active compounds on CDK2 when docked in EGFR had lower docking scores. Only one compound interacts with Lys721 (β-3 region) and Met769 (hinge region). The stability of interactions with CDK2 was assessed for 2k and 2l by molecular dynamics simulation using Desmond software. In conclusion, three compounds possess excellent activity against MCF-7 cell line and good activity against CDK2.
Ovulation Patterns Affect the Offspring Sex Ratios and Change With the Women’s Age
Authors: Fukuda Misao , Fukuda Kiyomi , Mason Shawn , Tatsumi Kenichi , Shimizu Takashi , Akahori Taiichiro , Matsumoto Tsunekazu , Tahaara Masahiro , Andersen Claus Y
Published: May 2021

Purpose The aim of this study was to evaluate whether women’s ages at conception and the ratio of male to female infants are associated with various ovulation patterns.Methods An observational clinical study was conducted in private OB/GYN clinics. Infertile women with regular menstrual cycles receiving intrauterine insemination (IUI) and/or in-vitro fertilization (IVF) had their ovulation patterns monitored in three consecutive spontaneous cycles receiving infertility treatment in the third cycle. Ovulation patterns were also observed in women with slight ovulation disorders during IUI and/or IVF in clomiphene citrate stimulated cycles. All the pregnant women’s ages at conception and their respective offspring sex ratios were compared to various ovulation patterns. Results Contralateral ovulation (i.e. ovulation jumping from ovary to the other) was more often observed in relatively younger women, who showed a higher probability of having a boy than after ipsilateral ovulation. There was a significantly higher frequency of boys being conceived following three consecutive ovulations with a left-left-right (LLR) ovulation pattern, while three ovulations from the left ovary (LLL) were associated with a higher frequency of girls. We also found two consecutive menstrual cycles the left-right (LR) ovulation pattern showed a similar significant difference compared to the left-left (LL) ovulation. Both the infertile and infertile + fertile women groups showing right-sided ovulation, regardless of age, showed significantly higher offspring sex ratio compared to left-sided ovulation, which was not observed in the group of fertile women alone. Conclusions LLR, LR and contralateral ovulation happens more often in younger women and favors male offspring in infertile women. Right-sided ovulation favors male offspring in infertile and infertile + fertile women, which was not observed in the group of fertile women.
Streptococcal Meningitis Reveals the Presence of Residual Streptococci and Down-Regulated Aquaporin 4 in the Brain
Authors: Nakayama Tatsuya
Published: May 2021

The pathology of streptococcal meningitis is poorly understood, even though streptococcal infection induces meningitis. The aim of this study was to clarify the relationship between streptococcal meningitis and aquaporin 4 (AQP4) in the mouse brain. After Streptococcus suis infection, the streptococcal number was calculated, and AQP4 mRNA expression in the brain was quantified at 2 and 7 days after infection. At 7 days post-infection, mice with neurological symptoms showed significantly higher S. suis levels in the brain than mice without neurological symptoms. AQP4 expression was significantly decreased in mice with neurological symptoms than in mice without neurological symptoms. Image analysis demonstrated that S. suis progressed to invade the white matter. Pathological analysis revealed that infected mouse brains had higher inflammation and neurological damage scores than uninfected mouse brains. Therefore, mice with neurological symptoms caused by streptococcal meningitis had high S. suis levels in the brain and reduced AQP4 expression.
The Association Between SYNTAX Score and Long-term Outcomes in Patients With Unstable Angina Pectoris: a Single-center Retrospective Study
Authors: Xu Min , Li Hong-Wei , Chen Hui
Published: May 2021

Background: SYNTAX Score affects clinical outcomes in early studies, whether SYNTAX Score could predict long-term outcomes in patients with unstable angina pectoris (UAP) in the era of new generation drug-eluting stent was unclear, and differences by SYNTAX Score in long-term outcomes between the medical therapy and percutaneous coronary intervention (PCI) in UAP patients were not well known.Methods：In this single-center retrospective study, a total of 2,364 patients with UAP from January 2014 to June 2017 at Beijing Friendship Hospital were enrolled. The primary endpoint was a composite of major adverse cardiovascular events (MACE) , including all-cause death, cardiac death, nonfatal myocardial infarction (MI) and stroke after at least 2 year from discharge. Results：In this study, 1,695 patients had low SYNTAX score (<22) , 432 patients had medium SYNTAX score (22-32), and 237 patients had high SYNTAX score (≥33), and 1,018 received medical therapy, 1,346 patients underwent PCI. Long-term MACE occurred in 95 patients during 3.38 ± 0.99 years follow up. Cox multivariate regression analysis showed advanced age, diabetes mellitus, heart failure, chronic kidney disease (CKD) and high SYNTAX score were independent predictors for MACE in the medical therapy group (P < 0.05), while, heart failure and CKD were predictors of MACE in PCI group. Compared to medical therapy group, PCI group showed lower MACE and cardiac death in patients with high SYNTAX score (≥33) (7.4% vs. 16.7%, P = 0.048; 3.7% vs 14.6%, P = 0.004), but no reduction in patients with low- and medium SYNTAX score.Conclusions：High SYNTAX score could predict long-term MACE for UAP patients with medical therapy, but not for patients undergoing PCI. Compared to medical therapy, PCI could significantly reduced long-term MACE and cardiac death for patients with high SYNTAX score.
Spin excitations in metallic kagome lattice FeSn and CoSn
Authors: Xie Yaofeng , Chen Lebing , Chen Tong , Wang Qi , Yin Qiangwei , Stewart J. , Stone Matthew , Daemen Luke , Feng Erxi , Cao Huibo , Lei Hechang , Yin Zhiping , MacDonald Allan , Dai Pengcheng
Published: May 2021

In two-dimensional (2D) metallic kagome lattice materials, destructive interference of electronic hopping pathways around the kagome bracket can produce nearly localized electrons, and thus electronic bands that are flat in momentum space. When ferromagnetic order breaks the degeneracy of the electronic bands and splits them into the spin-up majority and spin-down minority electronic bands, quasiparticle excitations between the spin-up and spin-down flat bands should form a narrow localized spin-excitation Stoner continuum coexisting with well-defined spin waves in the long wavelengths. Here we report inelastic neutron scattering studies of spin excitations in 2D metallic Kagome lattice antiferromagnetic FeSn and paramagnetic CoSn, where angle resolved photoemission spectroscopy experiments found spin-polarized and nonpolarized flat bands, respectively, below the Fermi level. Although our initial measurements on FeSn indeed reveal well-defined spin waves extending well above 140 meV coexisting with a flat excitation at 170 meV, subsequent experiments on CoSn indicate that the flat mode actually arises mostly from hydrocarbon scattering of the CYTOP-M commonly used to glue the samples to aluminum holder. Therefore, our results established the evolution of spin excitations in FeSn and CoSn, and identified an anomalous flat mode that has been overlooked by the neutron scattering community for the past 20 years.
Mechanistic Insights into Dideoxygenation in Gentamicin Biosynthesis
Authors: Li Sicong , Bury Priscila , Huang Fanglu , Guo Junhong , Sun Guo , Reva Anna , Huang Chuan , Jian Xinyun , Li Yuan , Zhou Jiahai , Deng Zixin , Leeper Finian , Leadlay Peter , Dias Marcio , Sun Yuhui
Published: May 2021

Gentamicin is an important aminoglycoside antibiotic used for treatment of infections caused by Gram-negative bacteria. Although most of the biosynthetic pathway of gentamicin has been elucidated, a remaining intriguing question is how the intermediates JI-20A and JI-20B undergo a dideoxygenation to form gentamicin C complex. Here we show that the dideoxygenation process starts with GenP-catalyzed phosphorylation of JI-20A and JI-20Ba. The phosphorylated products are converted to C1a and C2a by concerted actions of two PLP (pyridoxal 5'-phosphate)-dependent enzymes: elimination of water and then phosphate by GenB3 and double bond migration by GenB4. Each of these reactions liberates an imine which hydrolyses to a ketone or aldehyde and is then re-aminated by GenB3 using an amino donor. Crystal structures of GenB3 and GenB4 have guided site-directed mutagenesis to reveal crucial residues for the enzymes' functions. We propose catalytic mechanisms for GenB3 and GenB4, which shed new light on the already unrivalled catalytic versatility of PLP-dependent enzymes.
Feasibility Study on Machining Super-dimensioned Aviation Deep-hole Ti6Al4V Part With Axial Ultrasonic Vibration-assisted Boring
Authors: Sui He , Zhang Lifeng , Wang Shuang , Gu Zhaojun
Published: May 2021

Deep-holes are typical super-dimensioned parts of aircraft structures, with associated machining technology which is recognized as challenging due to the difficult-to-machine material, the narrow and enclosed machining environment, and the weak rigidity and large deformation of the boring bar. Axial ultrasonic vibration-assisted cutting has been proved to greatly enhance machining performance, especially in the precision machining of aviation alloy. This paper focuses on the machining of a super-dimensioned titanium alloy Ti6Al4V aviation deep-hole part (aspect ratio exceeding 20) with the axial ultrasonic vibration-assisted boring (AUVB) method. First, the kinetics of the AUVB process is analyzed and a retrospective of its separation cutting feature is provided. Subsequently, a multi-step cantilever beam model of the boring bar is established to analyze its static rigidity and dynamic stability. The aperture error of bored hole is deduced, and it is found to be mainly determined by the diameter of the basic hole, the cutting depth, the boring force and the length, diameter, elastic modulus, and rotational inertia of each step. Size coefficient, rather than aspect ratio, is then put forward to represent the static rigidity of the boring bar, which is proportional to the third power of the diameter and inversely proportional to the fourth power of the diameter. In addition, two different vibration cases, namely modal-coupling vibration and regenerative vibration are considered for dynamic stability analysis. The actual dynamic rigidity of AUVB in both of these cases is larger than that of conventional boring (CB), resulting from the lower actual boing force and the larger actual cutting depth. Next, the morphology of bored surface is analyzed, and the geometric height of peaks formed by AUVB and CB are calculated. Phase shift φ= π is suggested for obtaining a better surface in AUVB. Finally, the feasibility of AUVB on machining a super-dimensioned titanium alloy Ti6Al4V aviation deep-hole part with three different sizes of boring bar (size coefficients are 7473, 4076 and 2718, respectively) is verified through systematic experiments, and compared with CB. Results demonstrate that AUVB has obvious advantages in reducing the boring force, improving boring accuracy, suppressing vibration and promoting surface quality. Furthermore, the aperture error decreases to 50% and vibration amplitudes decrease to only 20%-25%. The overall surface roughness of the deep-hole part stays below Ra=0.8μm with rotational speeds of 60r/min and 80r/min, and the surface residual stress state is transferred from the tensile state to a compressive one. As a result, not only AUVB can provide better boring accuracy and surface finish, but it also can enhance the surface fatigue properties.
Ceria Nanoparticles Ameliorate Renal Fibrosis by Modulating the Balance Between Oxidative Phosphorylation and Aerobic Glycolysis
Authors: mengling Wang , feng Zeng , fengling Ning , yinhang Wang , jiaqi He , shilin Zhou , cong Li , dongliang Zhang , ping Hu , hong Xin , xudong Xu , xuemei Zhang
Published: May 2021

Background and aims: Renal fibrosis is the common outcome in all progressive forms of chronic kidney disease. Unfortunately, the pathogenesis of the renal fibrosis remains largely unexplored, among which metabolic reprogramming plays an extremely crucial role in the evolution of renal fibrosis. Ceria nanoparticles have strong ROS scavenging and anti-inflammatory activity. The present study aims to determine whether CeNPPEG has therapeutic value for renal fibrosis.Methods: Ceria nanoparticles and Ceria nanoparticles-PEG were synthesized according to previously reported procedures with slight modifications. For the in vivo studies, unilateral ureteral obstructive -induced kidney fibrosis was chosen. Through cellular studies, TGF-β1-induced epithelial-to-mesenchymal transition in HK-2 cells of enhanced glycolysis was initially utilized to report the associations among Ceria nanoparticles-PEG, oxidative phosphorylation, glycolysis and renal fibrosis.Results: In this study, Ceria nanoparticles were developed and the Ceria nanoparticles-PEG treatment significantly ameliorated renal fibrosis in unilateral ureteral obstructive mice as well as protected HK-2 cells from transforming growth factor beta1 -induced epithelial-to-mesenchymal transition process. Furthermore, we found that Ceria nanoparticles-PEG suppressed hexokinase 1and hexokinase 2 to block the dysregulated metabolic status, in which damaged epithelial cells take glycolytic metabolism as priority to oxidative phosphorylation, exerting the protective function in the progress of renal fibrosis.Conclusions: The application of Ceria nanoparticles-PEG in this study is attempted to provide another therapeutic option on renal fibrosis.
Sofosbuvir Is the Major Contributor to Elevation of Blood Cholesterol and Low-density Lipoprotein During HCV Treatment With DAAs
Authors: Wang Yi-Kai , Lee Wei-Ping , Wang Ying-Wen , Lu Chia-Ling , Huang Yi-Hsiang , Hou Ming-Chih , Chang Yuh-Lih , Lan Keng-Hsin
Published: May 2021

Background and aims: Worsened lipid profiles were observed in chronic hepatitis C (CHC) patients during direct-acting antivirals (DAAs) treatment, among which combination drugs confounded the effect of individual ingredient on lipid. Tenofovir alafenamide (TAF) also worsened lipid profiles in HIV patients. Structural similarity between sofosbuvir (SOF) and TAF prompted us to investigate rapid increase in total cholesterol (TC) and low-density lipoprotein (LDL) in CHC patients treated with SOF-based DAAs.Methods: A retrospective study was performed to analyze 487 CHC patients receiving DAAs with SVR12. Laboratory data were analyzed by logistic regression to determine relative risks of TC and LDL of SOF-based regimens over non-SOF-based regimens. Week 4 to baseline ratios of serum TC or LDL between regimens were compared by Mann-Whitney's test.Results: 487 patients were treated with Harvoni® (SOF-based, 206 patients), Epclusa® (SOF-based, 124 patients), Maviret® (non-SOF-based, 122 patients), or Zepatier® (non-SOF-based, 35 patients). At week 4 during drug treatment, Harvoni®, Epclusa®, and Maviret® induced statistically significant elevation of TC and LDL, but Zepatier® did not. SOF-based regimens had 2.72-fold higher relative risk (RR) causing 10% elevation of TC (95% CI: 1.84-4.02, p < 0.001) and 2.04-fold higher RR causing 10% elevation of LDL (95% CI:1.39-3.01, p < 0.001) than non-SOF-based DAAs. Conclusion: SOF-based DAAs were associated with significantly larger amplitude of increases in TC and LDL than non-SOFO-based regimens during the initial 4 weeks of treatment, but the amplitude was not sustained to SVR12.
Pseudomonas nanhaiensis sp. nov., a lipase-producing bacterium isolated from deep-sea sediment of the South China Sea
Authors: Pang Yu , Zhang Yage , Chen Mengru , Lu Wei , Chen Ming , Yan Yongliang , Lin Min , Zhang Wei , ZHOU Zhengfu
Published: May 2021

A bacterial lipase producer, designated SCS 2–3, was isolated from deep-sea sediment of the South China Sea. Phylogenetic analysis based on the 16S rRNA sequences revealed that strain SCS2-3 belonged to the genus Pseudomonas and had 98.56% similarity with P. xinjiangensis NRRL B-51270T that the closest relative strain. MLSA using four protein-coding genes (dnaK, gyrA, recA, and rpoB) showed strain SCS 2–3 to branch separately from its most closely related type strains. ANI and in silico DDH values between strain SCS 2–3 and the closely related type strains of Pseudomonas species were less than 81.51% and 23.80%, respectively, both below the species delineation threshold. Genome comparison showed that strain SCS 2–3 shared 1875 core gene families with other eight closely related type strains in Pseudomonas, and the number of strain-unique genes was 263. Through gene annotations, genes related to lipase was found in the isolates’ genomes. Furthermore, a combination of phenotypic, chemotaxonomic, phylogenetic and genotypic data clearly indicated that strain SCS 2–3 represents a novel species of the genus Pseudomonas, for which the name Pseudomonas nanhaiensis sp. nov. is proposed. The type strain is SCS 2-3T (= GDMCC 1.2219T = JCM 34440T).
Liposuction in Cancer‐Related Lower Extremity Lymphedema: an Investigative Study for Clinical Applications
Authors: Xin Jianfeng , Sun Yuguang , Xia Song , Chang Kun , Li Yiyin , Dong Jian , Shen Wenbin
Published: May 2021

Background: Lymphedema is a progressive, noncurable condition consisting of increases in subcutaneous fat and interstitial fluid in the limbs and fibrosis during later stages. The disease most commonly affects the limbs because of injury to or removal of lymph nodes. The aim of this study was to investigate therapeutic effects of liposuction for cancer-related lower extremity lymphedema.Methods 62 patients with cancer-related lymphedema in unilateral lower extremity were recruited in this study, and all the patients underwent liposuction. The volume of hemorrhage and lipid, the operation time and the volume changes of affected extremity were compared by applying t-test, and the subjective evaluation of the patients were also assessed by applying Chi-square. Results Total lipid volume is (2539±1253.5) ml, and the hemorrhage volume is (828±311.8) ml during liposuction. For the comparison of objective indexes, (1) Comparison of percent volume difference before surgery, in-operation and 3-month follow-up was (5.5±12.2 Vs. 11.6±18.4 Vs. 43.2±23.7, P＜0.05) respectively. (2) Higher lipid volume and liposuction rate for female patients, with lower volume of hemorrhage. (3) Higher volume of hemorrhage in patients with erysipelas (4) Higher lipid volume and liposuction rate, with lower hemorrhage for stage II than stage III patients.Conclusions Liposuction is an effective therapy for cancer-related lower extremity lymphedema, and it is necessary to combine with other therapeutic methods to improve lymph circulation.
Earthworms drastically change fungal and bacterial communities during vermicomposting of sewage sludge
Authors: Domínguez Jorge , Aira Manuel , Crandall Keith A. , Pérez-Losada Marcos
Published: May 2021

Wastewater treatment plants produce hundreds of million tons of sewage sludge every year all over the world. Vermicomposting is well established worldwide and has been successful in processing sewage sludge, which can contribute to alleviate the severe environmental problems caused by their disposal. Here, we utilized 16S and ITS rRNA high-throughput sequencing to characterize bacterial and fungal community composition and structure during the gut- and cast-associated processes (GAP and CAP, respectively) of vermicomposting of sewage sludge. Bacterial and fungal community composition and diversity changed significantly during both vermicomposting processes. Most of the bacterial and fungal taxa in the sewage sludge were eliminated during vermicomposting, mainly through the GAP. Vermicomposting of sewage resulted in a stable and rich microbial community with potential biostimulant properties that may aid plant growth. Our results support the use of vermicompost derived from sewage sludge for sustainable agricultural practices.
Development of the principles of fuzzy rule-based system for hepatocelular carcinoma staging
Authors: Mammadova Masuma , Bayramov Nuru , Jabrayilova Zarifa
Published: May 2021

The article proposes the principles for the development of a fuzzy rule-based physician decision support system n to determine the stages of the most common hepatocellular carcinoma (HCC) among malignant tumors of liver. The stages of HCC, i.e., critical situations, are expressed by different combinations of clinical signs of input data and emerging clinical conditions. These combinations shape the multiplicity of possible situations (critical situations) by forming linguistic rules that are in fuzzy relations with one another. The article presents the task of developing a fuzzy rules-based system for HCC staging by classifying the set of possible situations into given classes. In order to solve the problem, fuzzy rules of clinical situations and critical situations deviated from them are developed according to the possible clinical signs of input data. The rules in accordance with the decision-making process are developed in two phases. In the first phase, three input data are developed: nine rules are developed to determine possible clinical conditions based on the number, size, and vascular invasion of tumor. In the second phase, seven rules are developed based on possible combinations of input data on the presence of lymph nodes and metastases in these nine clinical conditions. At this stage, the rules representing the fuzzification of results obtained are also described. The latter provide an interpretation of results and a decision on related stage of HCC. It also proposes a functional scheme of fuzzy rules-based system for HCC staging, and presents the working principle of structural blocks. The fuzzy rule-based system for HCC staging can be used to support physicians to make diagnostic and treatment decisions
Acute hemolytic transfusion reaction caused by anti-Coa
Authors: Covin R.B. , Evans K.S. , Olshock R. , Thompson H.W.
Published: May 2021

Coa is a high-frequency blood group antigen in the Colton blood group system expressed on red blood cells (RBCs) of approximately 99.8 percent of random persons. Anti-Coa has been reported to cause delayed hemolytic transfusion reactions, hemolytic disease of the newborn, and accelerated clearance of RBCs in vivo. Acute hemolytic transfusion reactions (AHTRs) have not previously been reported. A 58-year-old man was hospitalized for vascular surgery. Initial blood bank evaluation revealed anti-Fya. The patient received six units of RBCs during his initial hospitalization and developed anti-E. A subsequent sample was sent to the reference laboratory when all units of RBCs appeared incompatible. Additional studies, including alloadsorptions, revealed the presence of anti-E, anti-Fya, and an apparent warm autoantibody. One unit of least-incompatible RBCs was transfused during surgery. The patient had an increase in temperature. Hemoglobinuria and a decrease in hematocrit were also noted. Due to the clinical impression of an AHTR, the pre- and postreaction samples were reevaluated in the reference laboratory and demonstrated the presence of anti-Coa in both. Based on clinical and laboratory evaluation this patient appears to have had an AHTR due to anti-Coa. This is the first known reported case of an AHTR caused by anti-Coa.
Immunohematology 2001;17:45–49.
DNA from urine sediment or buccal cells can be used for blood group molecular genotyping
Authors: Rios M. , Cash K. , Strupp A. , Uehlinger J. , Reid ANDM
Published: May 2021

Accurate blood group antigen typing of red blood cells with a positive direct antiglobulin test or from a recently transfused patient has been a long-standing problem. To overcome this problem, we evaluated the feasibility of using somatic cells as a source of DNA for molecular genotyping. Two sources of cells that could be obtained by noninvasive procedures were chosen for analysis: urine samples, which were already available in the clinical laboratory, and buccal epithelial cells collected with cotton wool swabs. DNA, prepared using a commercial kit, was subjected to polymerase chain reaction amplification and followed by digestion with the appropriate restriction enzyme. Genotyping was performed for three alleles encoded by polymorphic genes on three different chromosomes, namely KEL1/KEL2, JKA/JKB, and FYA/FYB. Genotyping results were compared to the results of typing performed on red blood cells using standard hemagglutination techniques. Results given by samples freshly collected from volunteer donors were concordant. Although results obtained with samples collected from hospital patients were initially not in agreement with the phenotyping results, adjustments to the test protocol resulted in concordance. DNA from blood, urine sediment, or buccal cells can be used for blood group molecular genotyping.
Immunohematology 15;2:61–65.
Case report: passively acquired anti-D in a D+ pregnant patient
Authors: Holub M.P. , Kitchen K. , Mensinger E.
Published: May 2021

A sample was submitted for serologic evaluation from a pregnant patient with immune thrombocytopenic purpura (ITP) for possible transfusion in the future because of a decreased platelet count. Anti-D and -E were identified in the patient’s serum using several antibody identification techniques, and anti-D was recovered in an acid eluate prepared from the patient’s red cells. It was discovered that WinRho™ had been administered to treat the ITP. This product has been licensed for treatment of nonsplenectomized D+ children and adults with ITP to increase the platelet count. Administration of anti-D to D+ individuals for treatment of ITP can cause a red cell anemia.
Immunohematology 1999;15:69–70.
Provision of HPA-1a (PlA1)-negative platelets for neonatal alloimmune thrombocytopenia: screening, testing, and transfusion protocol
Authors: Munizza M. , Nance S. , Keashen-Schnell M.A. , Sherwood W. , Murphy s.
Published: May 2021

HPA-1a-negative platelet products are not routinely available for newborns with alloimmune thrombocytopenia. In this article we describe a program established to identify normal pheresis donors who are HPA-1a-negative and to organize their future donations so that our regional blood center would always have an HPA-1a-negative platelet product available. The solid phase red cell adherence assay was used for initial screening of platelet pheresis products. HPA-1a-negative donors were confirmed with the platelet suspension immunofluorescence test using three anti-HPA-1a sera. Screening of 2600 platelet-pheresis donor samples identified 40 HPA-1a-negative donors. Of these, 36 are active and are coded for recognition on the daily pheresis inventory sheet. Theoretically, assuming four donations per year and donors’ cooperation with scheduling, these 36 donors would enable us to have at least one HPA-1a-negative product available every day. In addition, a decision tree for patient management using platelet serology and availability of HPA-1a-negative products was developed. The GTI-PAK™12 is the major technique used for serologic screening of mothers of patients thought to have neonatal alloimmune thrombocytopenia. By screening pheresis donors and developing a clinical decision tree, HPA-1a-negative products, a rare resource, can be fully utilized.
Immunohematology 1999;15:71–74.
A comparison of a new affinity column system with a conventional tube LISS-antiglobulin test for antibody detection
Authors: Shirey R.S. , Boyd J.S. , Barrasso C. , King K.E. , Ess P. M . N
Published: May 2021

A recently introduced system for antibody detection (ReACT™) consists of affinity columns (AFC) that contain protein A and protein G-coated agarose. We compared the ReACT™ system to a conventional tube low-ionic-strength saline antiglobulin test (LISS-AGT). We selected 100 LISS-AGT positive samples with clinically important and benign antibodies of varying strengths and 130 LISS-AGT negative samples to evaluate by the AFC method. AFC tests were positive with all 84 clinically important antibodies, including 36 antibodies that reacted ≤ 1+ at LISS-AGT (0% falsely negative). Eleven of 16 (69%) clinically benign antibodies reacted by AFC. Five samples (2 anti-Sda, 2 anti-I, and 1 inconclusive) were negative by AFC. AFC tests were negative with all 130 samples that were negative by LISS-AGT (0% falsely positive). The AFC method showed results comparable with results obtained with a conventional tube LISS-AGT for detection of clinically important antibodies. Some unwanted, clinically benign antibodies may not be detected by the AFC method.
Immunohematology 1999;15:75–77.
Serologic and molecular investigations of a chimera
Authors: Mifsud Nicole A. , Haddad Albert P. , EHart Cathie , Holdsworth Rhonda , Condon Jennifer A. , Swain Michael , Sparrow Rosemary L
Published: May 2021

A chimeric individual possesses two or more genetically distinct ceil populations. Although the chimerism may not be evident in all gene systems, various loci display greater numbers of alleles than genetically “normal” individ uals. The proposit’a was referred for further laboratory investigation due to a mixed-field ABO blood group reaction following routine antenatal testing. Various molecular (HLA class II, ABO genotyping, and 10 short tandem repeat [STR] microsatellites) and serologic (HLA class I and red cell blood groups) typing techniques were employed to investigate a number of polymorphic loci located on different chromosomes. Chimerism was identified in 8 out of the 1.4 chromosomes tested: chromosome 1 (Duffy), 6 (HLA class I and Iï), 9 (ABO), 11 (HUMTH01), 12 (HUMPLA2A1), 15 (HUMFES/FPS). 18 (Kidd) and 21 (D21S11). The proposita was determined to be a probable dispermic chimera, based on the results of the serology and molecular studies.
Precipitation of serum proteins by polyethylene glycol (PEG) in pretransfusion testing
Authors: Hoffer Jack , Koslosky William P. , Gloster Elizabeth S. , Dimaio Tberese M. , Reid Marion E.
Published: May 2021

Polyethylene glycol (PEG) is used as a potentiator of blood group antigen-antibody interactions. Although PEG is known to precipitate immunoglobulins, we could find no reports of this reagent entrapping red blood cells (RBCs) in irreversible clumps. The patient we describe here had hyperglobulineinia with a reversed albumin:giob- ulin ratio and a diffuse immunoglobulin peak on serum protein electrophoresis. During preparation of serologic tests, a precipitate formed that entrapped the RBCs when PEG was added. Rapid recognition of this phenomenon could prevent delay in the selection. of blood for transfusion by substituting PEG-indirect antiglobulin test (1AT) with another technique such as low-ionic-strength solution (LISS-IAT, and by increasing the number of washes prior to addition of the antigiobulin reagent.
Hemochromatosis, iron, and blood donation: a short review
Authors: Fields Angela C. , Grindon Alfred J.
Published: May 2021

Hereditary hemochromatosis (HIT), an autosomal recessive disease of iron overload, is one of the most common inherited diseases. The candidate gene (HFE) for HH lias been identified recently and a DNA- based test for the mutation is available. Treatment for HH patients with elevated iron stores include repeated phlebotomy. Left untreated, iron overload can lead to cirrhosis, organ failure, and a shortened life expectancy. In the past and present, blood collected for therapeutic purposes from patients with HH has been discarded. The aim of this article is to address whether blood collected from HH patients should be used for allogeneic transfusion in the future.
Use of LOR-15C9 monoclonal anti-D to differentiate erythrocytes with the partial DVI antigen from those with other partial D antigens or weak D antigens
Authors: Reid M.E. , Halverson G.R. , Roubinet F. , Apoil P.A. , Blancher A.
Published: May 2021

Historically, red blood cells (RBCs) with partial D antigens have been defined serologically by their pattern of reactivity with polyclonal and monoclonal anti-D. Although numerous variants have been described in tests with well-characterized monoclonal anti-D, definition remains difficult to ascertain serologically. RBCs of known partial D type were tested with LOR-15C9 (a monoclonal anti-D) and commercial anti-D by the tube indirect antiglobulin test (IAT), by micro typing system IgG gel cards, and by immunoblotting. By IAT, LOR-15C9 reacted strongly with DIIIa, DIIIc, DVa, DVI, DVII, and DFR RBCs in addition to RBCs with common D antigens; weakly with DII, DNU, and DIIIb RBCs; and not at all with DIVa, DIVb, DBT, or R0
Har RBCs. Reactivity was variable (1+ to 4+), with RBCs classified as weak D (Du). As expected, the commercial anti-D agglutinated all D variants and weak D RBC samples by the IAT and by using IgG gel cards; however, the reactivity with DVI RBCs was weaker than with LOR- 15C9. By immunoblotting, LOR-15C9 detected a band with an apparent molecular mass of approximate M
r 30,000-34,000 in membranes prepared from D-positive, DIIIa, DIIIc, DVa, DVI, DVII, and DFR RBCs and an additional band of Mr 20,000-22,000 in membranes prepared from DVI RBCs. No band(s) was detected in membranes from DII, DNU, DIIIb, DIVa, DIVb, DBT, R0
Har, weak D, or D-negative samples. LOR-15C9 provides a useful tool to identify positively DVI samples and thereby differentiate this partial D from other D variants and from weak D samples.
Use of the MAIEA assay to demonstrate that Fy3 is on the same glycoprotein as Fy6, Fya, and Fyb
Authors: Tzeng J. , Dodd R. , Mallory D.
Published: May 2021

The monoclonal-antibody immobilization of erythrocyte antigens (MAIEA) assay is a technique that detects trimolecular complexes formed by a human antibody and a mouse monoclonal antibody with specific red cell epitopes. This enzyme-linked immunoadsorbent assay test gives a positive reaction when two different epitopes on the same membrane protein are separately recognized by human and mouse antibodies. In this study, the MAIEA test was used to determine if the Duffy system antigen Fy3 is on the same membrane protein as Fy6. The well-established location and relationship of Duffy blood group antigens Fya, Fyb, and Fy6 were again confirmed by the MAIEA assay, and those facts were used to standardize a variation of the assay to establish the relationship between Fy3 and Fy6 using red cells with various Fy phenotypes. The MAIEA assay generated high absorbance values when Fy6 and Fya (or Fyb) antigens were evaluated. Similarly, high absorbance values were seen when Fy3 and Fy6 antigens were tested using Fy(a+b+), Fy(a+b–), and Fy(a–b+) red cells. The MAIEA assay is a valid technique for detecting Duffy system antigens. By this technique, it was shown that Fy3 and Fy6 antigens are carried on the Duffy glycoprotein D and by extension that Fy3 is also located on the Duffy glycoprotein D.
Detection of anti-D following antepartum injections of Rh immune globulin
Authors: Kennedy M.S. , McNanie J. , Waheed A.
Published: May 2021

Antepartum prophylaxis using Rh immune globulin (RhIG) at 28 weeks of gestation is routine in unsensitized Rh-negative women. As various sources state that anti-D may be detected up to 6 months after administration, we reviewed the medical and laboratory records of all Rh-negative women who delivered at our institution during 1995. For 385 evaluable women, only 137 (35.6%) had anti-D demonstrable in their sera at delivery; 97.8 percent of these delivered within 75 days after administration of RhIG. Of 248 women (64.4%) who delivered in < 76 days after administration of RhIG, 134 (54%) had demonstrable anti-D. For 123 women who delivered between 76 to 95 days after RhIG, only 3 (2.4%) had demonstrable anti-D. Of 14 women who delivered more than 96 days after RhIG, none had anti- D at delivery. These data show that the 300-μg dose used in the United States may not be adequate for antepartum protection and that the detection of anti-D more than 100 days after the administration of RhIG should be viewed with suspicion.
Evaluation of column technology for direct antiglobulin testing
Authors: Moulds J.M. , Diekman L. , Wells T. D.
Published: May 2021

Preliminary reports have suggested that microcolumn technology might be too sensitive for direct antiglobulin testing (DAT). We studied 228 samples from patients with autoimmune diseases and 30 samples from healthy controls to determine the sensitivity of column techniques. Both Sephadex® gel and protein A/G columns were compared with manual methods using rabbit or murine polyspecific reagents. Of the 187 samples that were negative by both manual methods, an additional 29 (15%) and 42 (22%) samples gave weakly positive reactions with the Sephadex® and protein A/G bead columns, respectively. Subsequently, there was poor correlation between manual and column techniques (r = 0.40–0.61). Acid eluates from these samples were negative. We concluded that the column technology may detect too many weakly positive DATs that are clinically insignificant.
Immune hemolytic anemia due to diclofenac
Authors: Laidlaw S.T. , Stamps R. , Booker D.J. , Brown M.J. , Sokol R.J.
Published: May 2021

A 37-year-old male presented with severe anemia, mild jaundice, and hemoglobinuria during his second course of diclofenac for gout. The peripheral blood showed microspherocytes and nucleated red blood cells (RBCs). The reticulocyte count was 21 percent and haptoglobin was < 0.1 g/L. A presumptive diagnosis of diclofenac-induced immune hemolysis was made and blood, urine, and drug samples were referred for investigation. Direct antiglobulin testing showed the RBCs to be coated with IgG1, IgG4, and C3d, but an eluate only yielded weakly reacting IgG antibodies. In tests for drug-dependent antibodies, group O, R1R2 red cells were incubated with the patient’s serum that had been mixed with either urine (which contained diclofenac metabolites) or diclofenac solution and then tested by an antiglobulin method. Strongly positive reactions with anti-IgG occurred in the tests using urine but only weak reactions in those tests employing diclofenac solution. All controls gave negative results. These findings support the role of diclofenac in causing hemolysis and the importance of employing urine as a source of drug metabolites. The findings also showed that an immune complex mechanism predominated and that the eluted IgG (detectable independently of the presence of the drug or its metabolites) confirmed a minor autoimmune component. Diclofenac was stopped and treatment with prednisolone and folic acid instituted; this resulted in complete recovery.
Immunohematology
1997;13:9–11.
Report on anti-Dib encountered in two Hong Kong Chinese
Authors: Lin C.K. , Mak K.H. , Chan N.K. , Yuen C.M.Y. , Devenish A. , Chan H.B. , Au K.L. , Szeto S.C.
Published: May 2021

Two cases of anti-Dib, a rarely encountered antibody, were identified in serum samples referred by hospital blood banks during the past 13 months. Case 1 is a 4l-year-old female who required blood for elective surgery. Case 2 is a premature infant suffering from mild neonatal jaundice on day 2 after birth. The anti-Dib in both cases exhibited marked dosage effect. The titer/score against Di(a+b+) and Di(a-b+) red blood cells (RBCs) in case 1 was 8/10 and 32/32, respectively, and in case 2, 4/18 and 32/46. The monocyte monolayer assay (MMA) also gave a similar pattern of results, being l5 percent and 100 percent reactive when tested with Di(a+b+) and Di(a-b+) RBCs in case 1, and 0.4 percent (within normal range) and l4.4 percent in case 2. The patient in case 1 underwent her operation without blood transfusion. The infant in case 2 was treated by phototherapy and subsequently recovered without the need for exchange transfusion.
Immunohematology
1997;13:17–19.
A modified PCR-RFLP genotyping method demonstrates the presence of the HPA-4b platelet alloantigen in a North American Indian population
Authors: Reiner A.P , Teramura G.
Published: May 2021

The diallelic HPA-4 (Pen/Yuk) platelet alloantigen system is polymorphic in Asian populations and accounts for the majority of cases of neonatal alloimmune thrombocytopenia in Japan. At the molecular level, the HPA-4a/4b dimorphism is associated with an arginine/ glutamine substitution at amino acid 143 of the gene encoding platelet glycoprotein IIIa. Unlike the five other major diallelic human platelet antigen systems (HPA-1, -2, -3, -5, and -6), the nucleotide substitution corresponding to the HPA-4 antigen system does not involve a common naturally occurring restriction enzyme site. This paper describes a new genotyping method for HPA-4 (polymerase chain reaction–restriction fragment length polymorphism [PCR-RFLP]) that involves restriction enzyme digestion of PCR-amplified genomic DNA using a modified PCR primer to create an artificial Taq I restriction site that is present in the HPA-4a but not in the HPA-4b DNA sequence. The HPA-4 PCR-RFLP method was validated by testing a reference panel of 10 known HPA-4 genotyped Japanese individuals. Thus, genotyping by PCR-RFLP can now be performed for all six major HPA systems. Using the HPA-4 PCR-RFLP genotyping method, we determined a frequency of 2.9 percent for the HPA-4b allele in a North American Indian population. This finding indicates the importance of the HPA-4 antigen system as a potential cause of alloimmune thrombocytopenia in American Indians.
Common variable immunodeficiency: diagnosis by absent ABO reverse type
Authors: Julius C.J. , Wade M. , Waheed A. , Mcneil D.L. , Kennedy M.S.
Published: May 2021

Many causes for ABO discrepancy between red blood cell and serum testing have been cited in the literature. ABO discrepancy due to weak or absent reverse type is most often seen at the extremes of age. We report a case of ABO discrepancy in a 25-year-old woman who presented for wisdom teeth extraction. Initial serologic workup revealed total absence of isohemagglutinin anti-B in this group A, D+ individual. Further evaluation revealed decreased levels of all classes of immunoglobulins and a medical history significant for multiple episodes of infection. Based on the patient’s history and laboratory data, she was diagnosed with common variable immunodeficiency (CVID). CVID can cause ABO discrepancy in an adult patient because of an absent ABO reverse type.
Immunohematology
1997;13:80–83.
Comparison of hemagglutination and solid phase titration methods for determination of critical prenatal antibody titers
Authors: Alder B.C. , Summers S.H. , Hare V.
Published: May 2021

The hemagglutination (HA) tube method is the standard method for determination of antibody titer in prenatal samples. Most facilities use a titer between 8 and 32 as their definition of a critical value when amniocentesis may be considered. This study determined if there is a relationship between the results of HA tube and solid phase (SP) titers performed on the same sample. Forty-six paired samples containing known antibody were titrated by both HA tube and SP methods and the results subjected to data analysis. We conclude that there is a strong correlation between HA tube and SP methods in titer end points; i.e., SP was greater than or equal to HA in every case. In addition, the appropriate critical SP titer is 64, which is at least equivalent to an HA critical titer of 16.
Immunohematology
1997;13:84–89.
The Role of Video in the Flipped Language Classroom
Authors: Verch Angelika , Nissen Elke
Published: May 2021

Flipped classrooms have become a widespread form of teaching. Yet, there is no consensus on how to define flipped (language) learning. Several authors consider the use of videos that prepares in-class activities as an essential principle.
The article presents a study which examined the actual roles of videos in a corpus of 52 descriptions by L2 teachers of flipped language class settings and using Willis’ 1983 framework.
In the corpus videos played a central role in before-class activities; a large number of videos were used. The roles that videos played in before-class activities in the settings did not all correspond to Willis’ framework; those which did not fit corresponded to direct instruction. The definition of a flipped setting was found to be unclear, as in a quarter of the descriptions the criteria did not apply. Video was not found to be necessarily constitutive for flipped language classes.
Serious Games in Higher Distance Education
Authors: Celestini Ann
Published: May 2021

Games have been socially entrenched throughout history as a form of entertainment. Current rapidly changing technological advances have permitted an increasingly prominent means of utilizing these sources of entertainment in an instructional capacity for educational purposes. Serious gaming as a result, focuses on engaging learners in activities which are not solely developed for enjoyment purposes. Goal oriented pursuits based in either an authentic or fictitious scenario can be designed to improve a learner or players motor and cognitive abilities or knowledge (de Freitas & Jarvis, 2006; Lamb et al., 2018; Protopsaltis et al., 2011). Serious gaming promotes intentional, active, and mobile learning that can be successfully used as a supplemental educational tool to facilitate a situated understanding of specific content (Admiral et al., 2011; Gee, 2005). This paper is a brief overview of game-based learning, or serious games, as an innovative instructional strategy in higher distance education.