Anti-E alloimmunization from a platelet apheresis transfusion in a 22-month-old male with acute myeloid leukemia.

RhD alloimmunization from platelet transfusions have been documented in the literature. However, non-RhD platelet alloimmunization is much less frequent and the risk for non-RhD alloimmunization from platelets is thought to be extremely low and most associated with buffy coat pooled platelets. A 22-month-old male with acute myeloid leukemia received 99 mL apheresis platelets for thrombocytopenia. Three months later, an antibody screen, the direct antiglobulin test (DAT), and red blood cell (RBC) genotype were sent for laboratory evaluation. The antibody screen was positive, with anti-E identified. The DAT was negative and the RBC genotype of the patient was predicted to be negative for the E antigen whereas the platelet donor was predicted to be positive for E antigen. There is a risk of alloimmunization of non-RhD antigen from platelet pheresis transfusion even in a patient less than 2 years old.

[Identification of Complex and Combined Antibody Consisted of Anti-c, Anti-E, Anti-Jka and Anti-Fya].

OBJECTIVE: To investigate the role of multiple serological methods in the identification of complex antibodies. METHODS: The blood group antigens were detected by saline and microcolumn agglutination methods. The saline method was used to screen and identify IgM-type antibodies in the patient's serum, while the polybrene, anti-globulin, microcolumn agglutination, enzymic and absorption-elution methods were used to screen and identify IgG-type antibodies. RESULTS: The patient was B/CCDee/Jk(a-b+)/Fy(a-b+) blood type. The serum reacted with panel cells, and the reaction presented anti-E pattern in the saline medium. It was fully positive in the microcolumn agglutination card, except 2 negative ones after using papain to treat the panel cells. Referring to the pattern table, it was concluded that there existed anti-c, anti-E, and anti-Jka antibodies, and one antibody corresponding to an antigen that was easily destroyed by papain. The red blood cells with specific phenotype were selected for absorption-elution to identify IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies. CONCLUSION: It is confirmed that IgM-type anti-E, and IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies exist in the patient's serum by multiple serological methods.

Low-cost goldleaf electrode as a platform for Escherichia coli immunodetection.

Gold electrodes are one of most prevalent substrates in electrochemical biosensors because they can be easily and highly efficiently functionalized with thiolated biomolecules. However, conventional methods to fabricate gold electrodes are costly, time-consuming and require onerous equipment. Here, an affordable method for rapid fabrication of an electrochemical immunosensor for Escherichia coli detection is presented. The gold electrode was generated using 24-karat gold leaves and lowcost polyvinyl chloride adhesive sheets covered with an insulating PTFE layer. The goldleaf electrode (GLE) was patterned using laser ablation and characterized by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electronic microscopy, contact angle and 3D profiling. The GLEs were modified by a self-assembled mercaptopropionic monolayer, followed by surface activation to allow binding of the specific anti-E. coli antibody via carbodiimide linking. The biosensor showed a detection limit of 2 CFU/mL and a linear dynamic range of 10-107 CFU/mL for E. coli cells. No false positive signals were obtained from control bacteria. The obtained results demonstrated suitability of GLE for use in biosensors with high reliability and reproducibility. It is foreseeable that our work will inspire design of point-of-need biosensors broadly applicable in low-resource settings.

Identification of immune hemolytic transfusion reaction and exploration of transfusion compatibility testing / 中国输血杂志

【Objective】 To analyze the causes of immune hemolytic transfusion reaction in one case, identify related antibodies, and explore transfusion compatibility testing. 【Methods】 ABO/Rh blood group identification, unexpected antibody identification of serum and diffusion fluid, direct antiglobulin test(DAT) and cross matching were conducted by saline method and/or microcolumn gel method. 【Results】 The patient′s blood group was O, and Rh phenotype was identified as DCCee. The DAT was negative, with strong anti-E antibody and weak anti-c antibody detected. Acute hemolytic transfusion reaction occurred in the patient after the last transfusion. 【Conclusion】 Currently, immune hemolytic transfusion reaction in China are mainly caused by Rh blood group system antibodies. The absence of unexpected antibody screening before blood transfusion and the weak anti-c antibody which resulted in missed detection of non compatibility in cross matching led to acute hemolytic transfusion reaction. It is recommended to conduct unexpected antibody screening before blood transfusion, and to collect blood sample for testing as soon as possible to improve the accuracy of DAT when acute hemolytic transfusion reaction is suspected.

Identification of Complex and Combined Antibody Consisted of Anti-c, Anti-E, Anti-Jka and Anti-Fya / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of multiple serological methods in the identification of complex antibodies.@*METHODS@#The blood group antigens were detected by saline and microcolumn agglutination methods. The saline method was used to screen and identify IgM-type antibodies in the patient's serum, while the polybrene, anti-globulin, microcolumn agglutination, enzymic and absorption-elution methods were used to screen and identify IgG-type antibodies.@*RESULTS@#The patient was B/CCDee/Jk(a-b+)/Fy(a-b+) blood type. The serum reacted with panel cells, and the reaction presented anti-E pattern in the saline medium. It was fully positive in the microcolumn agglutination card, except 2 negative ones after using papain to treat the panel cells. Referring to the pattern table, it was concluded that there existed anti-c, anti-E, and anti-Jka antibodies, and one antibody corresponding to an antigen that was easily destroyed by papain. The red blood cells with specific phenotype were selected for absorption-elution to identify IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies.@*CONCLUSION@#It is confirmed that IgM-type anti-E, and IgG-type anti-c, anti-E, anti-Jka and anti-Fya antibodies exist in the patient's serum by multiple serological methods.

Predictive Value of Anti- E6 Oncoprotein (High Risk- Human Papilloma Virus) and p16 Ink4a for Detecting HPV in Oral Epithelial Dysplasia.

AIM: The aim of this study was to evaluate the expression of human papillomavirus (HPV) in oral potentially malignant disorders (OPMD) and to examine the association of HPV in histological grades of dysplasia using p16 and Anti-E6 oncoprotein immunohistochemistry (IHC). SUBJECTS AND METHODS: This study focused on clinically diagnosed oral potentially malignant disorders. Clinical parameters such as age, gender, habits, occupation, duration, site, and the type of the lesions were examined and the incisional biopsy was done on the selected cases for the histopathological diagnosis. Selected cases of OPMDs were screened immunohistochemically for HPV 16 and HPV 18 (high-risk group) positivity using p16INK4a and Anti-E6 oncoprotein. The immunohistochemical p16 expression was evaluated based on (a) percentage of p16 positive cases and (b) pattern of p16 staining in various grades of OPMD. RESULTS: Anti-E6 oncoprotein (HR-HPV) expression level was only detected in 11 cases (37%), and positive expression of p16 was found in three cases (10%), with variation in cell proportion and intensity. Subsequently, the association between p16 expression level and clinicopathological characteristic factors was analyzed and a significant association was found between age and histopathology. CONCLUSION: There was an association between HPV and OPMD. Both biomarker tests, HPV E6 and p16 immunocytochemistry had a specific role in the detection of HR-HPV. Anti-E6 immunocytochemistry can be a valuable test with higher specificity for HPV DNA detection in oral epithelial dysplasia without losing sensitivity.

Hemolytic disease of the fetus and newborn caused by irregular antibody: A mortality case report and case series during the past 15 years in NCKUH.

OBJECTIVE: Hemolytic disease of the fetus and newborn (HDFN) caused by irregular antibodies is a rare, but possibly life-threatening condition. We report a case of severe intrauterine hemolysis caused by anti-E alloimmunization, and review 16 cases in the past 15 years of our hospital. CASE REPORT: A woman with gestational age 28 weeks and 5 days, received emergent Cesarean section because of fetal distress. The baby was expired at the next day after delivery and the comprehensive study showed severe anemia and alloimmunization related hemolysis caused by anti-E due to high antibody titer (1: 4096). CONCLUSION: Anti-E antibody is one of the most common non-Rhesus D antibodies in the pathogenesis of HDFN, but rarely leads to severe hemolysis. However, our case has the highest reported anti-E titer in HDFN and is the first case of mortality during the past 15 years in NCKUH.

Capture and identification of bacteria from fish muscle based on immunomagnetic beads and MALDI-TOF MS.

In the present study, E. coli was taken as a model bacterium, anti-E. coli functionalized magnetic beads were constructed and used to capture E. coli from aqueous extracts of fish sarcoplasmic protein (FSP) and fish muscle protein of sablefish. The excellency of the reproducibility of the present protocol was demonstrated by capturing E. coli from sablefish FSP extracts. The presence of 10 CFU/mL E. coli is still detectable. A microbial safety test on the surface of fish muscle was successfully performed. The bacterial identification accuracy from samples with different matrices was found to be excellent with RSD = 3%. High specific detection of target bacteria in complex biological samples was testified by spiking Staphylococcus aureus and Klebsiella pneumoniae in samples as interference. Ten biomarker ions were discovered for E. coli's recognition. It is promising to apply the present protocol in bacterial analysis in muscle food samples to ensure their safety.

Electrochemical Detection of Waterborne Bacteria Using Bi-Functional Magnetic Nanoparticle Conjugates.

Detection of microbial contamination in water is imperative to ensure water quality. We have developed an electrochemical method for the detection of E. coli using bi-functional magnetic nanoparticle (MNP) conjugates. The bi-functional MNP conjugates were prepared by terminal-specific conjugation of anti-E. coli IgG antibody and the electroactive marker ferrocene. The bi-functional MNP conjugate possesses both E. coli-specific binding and electroactive properties, which were studied in detail. The conjugation efficiency of ferrocene and IgG antibodies with amine-functionalized MNPs was investigated. Square-wave voltammetry enabled the detection of E. coli concentrations ranging from 101-107 cells/mL in a dose-dependent manner, as ferrocene-specific current signals were inversely dependent on E. coli concentrations, completely suppressed at concentrations higher than 107 cells/mL. The developed electrochemical method is highly sensitive (10 cells/mL) and, coupled to magnetic separation, provides specific signals within 1h. Overall, the bi-functional conjugates serve as ideal candidates for electrochemical detection of waterborne bacteria. This approach can be applied for the detection of other bacteria and viruses.

Ultrasonic-assisted preparation of eucalyptus oil nanoemulsion: Process optimization, in vitro digestive stability, and anti-Escherichia coli activity.

Eucalyptus oil (EO) is a natural and effective antimicrobial agent; however, it has disadvantages such as poor water solubility and instability. The aim of this study was to investigate the effect of process vessels and preparation process parameters on the particle size of the emulsion droplets using ultrasonic technique and response surface methodology to prepare eucalyptus oil nanoemulsion (EONE). The optimal sonication process parameters in conical centrifuge tubes were confirmed: sonication distance of 0.9 cm, sonication amplitude of 18%, and sonication time of 2 min. Under these conditions, the particle size of EONE was 18.96 ± 4.66 nm, the polydispersity index was 0.39 ± 0.09, and the zeta potential was -31.17 ± 2.15 mV. In addition, the changes in particle size, potential, micromorphology, and anti-Escherichia coli activity of EONE during digestion were investigated by in vitro simulated digestion. The emulsion was stable in simulated salivary fluid, tended to aggregate in simulated gastric fluid, and increased in particle size and potential value in simulated intestinal fluid. EONE showed higher anti-E. coli activity than EO by simulated digestion. These results provide a useful reference for the in vivo antimicrobial application of the essential oil.

Nanoparticle immuno-fluorescent probes as a method for detection of viable E. coli O157:H7.

Development of revolutionary sensitive biosensors for detecting the presence of harmful biological species in the environment is a necessity for countering disease outbreaks. This work examined the interaction of fluorescence-labeled antibody on amine functionalized gold nanoparticles (GNP) as a model system. The synthesized tetramethylrhodamine isothiocyanate (TRITC) labeled antibody-amine functionalized GNP interaction was characterized using UV-Vis spectroscopy and Fluorescent Microscopy imaging. Transmission Electron Microscopy (TEM) was also used to observe the morphology of the GNP. In contrast to TEM, the fluorescence microscopy imaging revealed the coating of the TRITC labeled antibody on the surface of the GNP. The signals were measured using a Photon Technology Inc. fluorometer at excitation of 541 nm and emission at 555 nm to 650 nm. Tests were conducted at near real-time with results obtained using the biosensor assay within 5 min. Results indicated that there was a shift of the wavelength from lower to higher wavelength (blue to red shift) when conjugated GNP (anti-E. coliO157:H7; IgY-TRITC-GNP) are compared to free GNP, a difference of about 28 nm. The GNP demonstrated a quenching capability when compared to the TRITC labeled antibody (degree of labeling of 15.41 mol dye per mole of IgY) using fluorometer. The lower and upper detection range of this method was found to be 103-105 CFU/mL with observed fluorescence of about 42,000 counts per seconds as against 24,000 counts per seconds that was observed when the specificity of the sensor was tested using Salmonella enterica.

Enfermedad hemolítica perinatal en recién nacido por aloinmunización anti-E / Foetal Haemolytic Disease in a new-born with Anti-E alloimmunization

AntecedentesLa aloinmunización anti-E es una etiología rara de enfermedad hemolítica perinatal (EHP). Su incidencia es muy baja, y menos del 3% es grave. Dado que la titulación de anticuerpos no se correlaciona con la gravedad, resulta interesante notificar casos que requieren tratamiento en el periodo postnatal.ObjetivoReportar el caso de un recién nacido tratado con fototerapia y exanguinotransfusión por EHP por aloinmunizacion anti-E.CasoMujer 42 años, escrutinio de anticuerpos irregulares positivo con aloanticuerpos anti-E.ConclusionesSe requiere un manejo activo tanto en el periodo prenatal como neonatal de la aloinmunización anti-E. Aunque no se haya documentado anemia en el periodo fetal, el recién nacido sigue siendo de alto riesgo para la presentación de clínica compatible con enfermedad hemolítica en el periodo postnatal.

BackgroundAnti-E alloimmunization is a rare aetiology of haemolytic disease of the foetus and new-born (HDFN). The incidence is low, with less than 3% with severity criteria. Antibody titre does not correlate with severity, so cases that require treatment should be notified.ObjectiveTo report the case of a new-born who required phototherapy as treatment of HDFN secondary to anti-E alloimmunization.Case42-year-old woman, positive irregular antibody screening, with anti-E alloantibodies.ConclusionsActive management is required in both the prenatal and the neonatal period. Although anaemia has not been documented in the foetal period, the new-born is still at high risk for the presentation of symptoms compatible with haemolytic disease in the postnatal period.

Is it reasonable for the use of Rh-ee blood? A hospital-based survey from a southern medical center in Taiwan.

Identification of alloantibodies and achieving a reduction in the rate of red blood cell (RBC) alloimmunization are important issues to prevent transfusion complications. The aim of this study was to identify the antigen and alloantibodies in our patients and to study the association of alloimmunization with previous transfusion. Transfusion records from the blood bank of Kaohsiung Medical University Hospital between 2015 and 2017 were retrospectively enrolled in the study. Antigen and antibody identification was performed using routine blood bank methods. In total, 56,422 transfusion records from 2015 to 2017 were included in the study. Among them, 1858 alloantibody episodes were found in the pre-transfusion survey, and anti-Mia, anti-E, and cold antibodies were the most common alloantibodies, with a prevalence of 3.29% (1858/56,422). Among them, 130 episodes involved newly found alloantibodies with no alloantibodies found in the previous transfusion survey. Tracing back to these newly transfusion-induced alloantibodies, the antibody was found with a mean of 10.8 ± 7.8 units of packed RBC transfusion, a mean of 66.3 ± 52.8 days, and with a mean of 4.3 ± 2.7 times of transfusion from the first transfusion therapy. An antibody survey revealed that Rh-ee (62.1%) was the most common phenotype in these newly identified antibodies. In summary, this hospital-based study revealed that RBC alloantibody rates were present at rates of 3.29%, with anti-Mia, anti-E, and cold antibodies being the most common alloantibodies. Among them, anti-E was the most commonly developed alloantibody. Given that the Rh-ee group is the most common phenotype in our population, the strategy of using Rh-ee blood for Rh-ee recipients is reasonable for transfusion safety.

Patient blood management in a neurosurgical patient with anti-e antibody.

The successful application of patient blood management approach in a 48-year-old neurosurgery patient planned for meningioma excision and requiring transfusion is described. The patient had multiple past immunizing events and developed antibody against a high-frequency antigen "e" of the Rh blood group system. With the joint effort from transfusion medicine specialist, anesthesiologist, and surgeon, the patient was successfully managed using the preoperative autologous blood donation program.

Faster and less invasive tools to identify patients with ileal colonization by adherent-invasive E. coli in Crohn's disease.

BACKGROUND AND AIMS: The identification of Crohn's disease (CD)-associated adherent and invasive Escherichia coli (AIEC) is time-consuming and requires ileal biopsies. We aimed to identify a faster and less invasive methods to detect ileal colonization by AIEC in CD patients. METHODS: CD patients requiring ileo-colonoscopy were consecutively enrolled in this prospective multicenter study. Samples from saliva, serum, stools, and ileal biopsies of CD patients were collected. RESULTS: Among 102 CD patients, the prevalence of AIEC on ileal biopsies was 24.5%. The abundance and global invasive ability of ileal-associated total E. coli were respectively ten-fold (p = 0.0065) and two-fold (p = 0.0007) higher in AIEC-positive (vs. AIEC-negative), while abundance of total E. coli in the feces was not correlated with AIEC status in the ileum. The best threshold of ileal total E. coli was 60 cfu/biopsy to detect AIEC-positive patients, with high negative predictive value (NPV) (94.1%[80.3-99.3]), while the global invasive ability (>9000 internalized bacteria) was able to detect the presence of AIEC with high positive predictive value (80.0% [55.2-100.0]). Overall, 78.1% of the AIEC + patients were colonized by two or less different AIEC strains. The level of serum anti-total E. coli antibodies (AEcAb) was higher in AIEC-positive patients (p = 0.038) with a very high negative predictive value (96.6% [89.9-100.0]) (p = 0.038) for a cut-off value > 1.9 × 10-3 . CONCLUSIONS: More than two thirds of AIEC-positive CD patients were colonized by two or less AIEC strains. While stools samples are not accurate to screen AIEC status, the AEcAb level appears to be an attractive, rapid and easier biomarker to identify patients with Crohn's disease harboring AIEC.

Development of Antibodies against HPV-6 and HPV-11 for the Study of Laryngeal Papilloma.

Laryngeal papilloma (LP), which is associated with infection by human papillomavirus (HPV)-6 or -11, displays aggressive growth. The precise molecular mechanism underlying the tumorigenesis of LP has yet to be uncovered. Building on our earlier research into HPV-6, in this study, the viral gene expression of HPV-11 was investigated by quantitative PCR and DNA/RNA in situ hybridization. Additionally, newly developed antibodies against the E4 protein of HPV-6 and HPV-11 were evaluated by immunohistochemistry. The average viral load of HPV-11 in LP was 1.95 ± 0.66 × 105 copies/ng DNA, and 88% of HPV mRNA expression was found to be E4, E5a, and E5b mRNAs. According to RNA in situ hybridization, E4 and E5b mRNAs were expressed from the middle to upper part of the epithelium. E4 immunohistochemistry revealed a wide positive reaction in the upper cell layer in line with E4 mRNA expression. Other head and neck lesions with HPV-11 infection also showed a positive reaction in E4 immunohistochemistry. The distribution pattern of HPV DNA, viral mRNA, and E4 protein in LP with HPV-11 infection was quite similar to that of HPV-6. Therefore, it might be possible to apply these E4-specific antibodies in other functional studies as well as clinical applications, including targeted molecular therapies in patients with HPV-6 and HPV-11 infection.

Formulation, physicochemical characterization, and anti- E. coli activity of food-grade nanoemulsions incorporating clove, cinnamon, and lavender essential oils.

This research studies the application of a specific nanoemulsion as anti-Escherichia coli agent. The specific mixture was generated by a simplex-centroid design. Physicochemical parameters such as droplet average diameter, pH, viscosity, density, turbidity, whitening index, refractive index, stability (thermal, physical, and osmotic stability), and antibacterial activity kinetic, have been assessed. The mixture nanoemulsions had droplet diameters significantly smaller than those of clove or cinnamon nanoemulsions. Individual and mixture essential oils nanoemulsion exhibited appropriate stability under pH, thermal, and ionic stress as well as after mid-term storage. Antibacterial activity kinetic revealed the fast and pronounced efficacy of mixture nanoemulsions on E. coli (reach 98% of growth inhibition), especially for the nanoemulsion composed of 50% essential oil in the dispersed phase upon 20 days of storage. All data considered, the actual work evidences the promising advantages of using specific nanoemulsions as delivery systems of antibacterial agents in the beverage and food industry.

Coordinated Expression of HPV-6 Genes with Predominant E4 and E5 Expression in Laryngeal Papilloma.

Laryngeal papilloma (LP) associated with human papillomavirus (HPV)-6 or -11 infection shows aggressive growth. However, the detailed molecular mechanism of virus-driven tumorigenesis has not been uncovered fully. HPV-6 viral gene expression and dynamic alterations were investigated with in situ localization of viral DNA and RNA in 13 patients with HPV-6-infected laryngeal papilloma. The average viral load was 4.80 × 105 ± 1.86 × 105 copies/ng DNA. E4, E5a, and E5b mRNAs accounted for 96% of the expression of 9 mRNAs. The alteration of viral DNA load during recurrence paralleled the mRNA expression levels, and the expression of all mRNAs showed a similar curve. E4, E5a, and E5b were expressed in the middle to upper part of the epithelium and were co-expressed in the same cells. E4 immunohistochemistry demonstrated an extensively positive reaction in the upper cell layer in accordance with E4 mRNA expression. These results suggest that individual viral genes are coordinately expressed for viral replication, virus release, and immunosurveillance avoidance. The newly developed E4-specific monoclonal antibody can be applied to further functional studies and clinical applications such as targeted molecular therapies.

Hemolytic Disease of the Fetus and Newborn Caused by Maternal Autoantibody with Mimicking Anti-E Specificity.

OBJECTIVE: There are few reports of hemolytic disease of the fetus and newborn (HDFN) caused by maternal autoantibodies. METHODS: We describe the case of a pregnant patient aged 26 years with systemic lupus erythematosus without any transfusion history who developed autoantibody with mimicking anti-E specificity. Her newborn developed HDFN caused by the maternal autoantibody. RESULTS: The clinical symptoms of the newborn were not serious. After bilirubin light phototherapy and other symptomatic supportive treatment, the baby was discharged with a good prognosis. CONCLUSION: This is the first reported case of HDFN caused by maternal autoantibody with mimicking anti-E specificity. However, the real antigenic target of the autoantibody was not clear.