Analysis of Correlation between Bone Marrow Vascular Endothelial Growth Factor per Platelet Count and Blast Percentage in Hematologic Malignancies.

OBJECTIVE: Bone marrow (BM) samples reflective of the BM condition are more suitable than peripheral blood samples for the measurement of vascular endothelial growth factor (VEGF) levels in relation to hematologic malignancy. However, BM VEGF levels require calibration, with respect to the platelet count, to eliminate any influence of VEGF released from platelets during sample processing. This parameter is termed the BM VEGF per platelet count. Our aim was to measure BM VEGF per platelet count of patients diagnosed with hematologic malignancy and to analyze its association with several hematological parameters, including BM blast percentages. We also compared the BM VEGF per platelet count and BM blast percentage between the disease and control groups. METHODS: BM plasma samples were collected from 73 patients classified into myeloproliferative neoplasm, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), plasma cell neoplasm (PCN), and control groups. Luminex assays were used to quantify BM VEGF levels. BM cellularity and cell percentage were assessed using BM biopsy and aspiration slides, respectively. Data on hematological parameters were collected from medical records. Single and multiple regression analyses were performed to analyze the relationships between different parameters. Intergroup comparisons were performed using the Kruskal-Wallis H-test. RESULTS: Independent factors associated with BM VEGF per platelet count included BM blast percentage (%), BM band neutrophil %, BM neutrophil %, and BM cellularity. Multiple regression analyses using the four factors generated the following model (adjusted R2=0.290): BM VEGF per platelet count (×10-6 pg)=1.488+0.059×BM blast (%). BM VEGF per platelet count was the highest in the AML group (n=13) and lowest in the control group (n=14). Similarly, BM blast % was the highest in the AML group. Compared to that in the PCN group (n=11), the BM blast % in the MDS group (n=12) was higher. CONCLUSION: BM VEGF per platelet count was related to BM blast %, and both parameters showed similar intergroup patterns, indicating an association.

Validity of Luminex and Enzyme-Linked Immunosorbent Assay Measuring Vascular Endothelial Growth Factor and Six Cytokines Quantitatively in Bone Marrow Aspiration Supernatant: Pilot Study.

OBJECTIVE: Vascular endothelial growth factor (VEGF) and other cytokines have been reported to be implicated in the molecular pathogenesis of hematologic malignancy. However, a quantitative measurement of VEGF and related cytokines is necessary to reflect the real situation in the bone marrow (BM). Currently, no such quantitative assays exist for use in the BM supernatant as their concentrations have not been previously validated in the BM. Here we performed linearity and recovery tests to quantitatively measure the concentrations of VEGF and six related cytokines in the BM. METHOD: A total of 24 BM supernatant samples were collected from patients who underwent a BM examination for hematological malignancies. The levels of VEGF and six cytokines - granulocyte colony-stimulating factor (G-CSF), interferon-ß (INF-ß), interleukin (IL)-1ß, IL-6, IL-17A, and tumor necrosis factor-α (TNF-α) - were measured using Luminex assay and enzyme-linked immunosorbent assay. Percentage recovery and linearity were calculated, with the acceptable range being 80-120%. The undiluted and diluted (1:2, 1:4, and 1:8) concentrations of VEGF and the six cytokines in 24 spiked and unspiked BM supernatant samples and controls were also measured. RESULTS: For VEGF, both assays passed the percentage recovery and linearity tests; wherein the undiluted and all diluted concentrations of VEGF in all six unspiked BM samples showed linearity parallel to those of VEGF in spiked BM samples and controls. For the other six cytokines, both assays did not pass the percentage recovery and linearity tests, with the undiluted and diluted concentrations in all seventeen unspiked BM samples (except G-CSF in one sample) showing a lack of parallelism to those in spiked BM samples and controls. CONCLUSIONS: Quantitative VEGF measurement in real BM specimens was validated using both Luminex assay and ELISA. All six cytokines, except for VEGF, whether undiluted or diluted, could not be accurately measured in the BM supernatants, indicating the presence of inhibitors to the analytes. Quantitative measurement of VEGF-related cytokines in the BM will have to be validated in further studies with more samples.

Analysis of the Relationship between the Expression Levels of Neutrophil Gelatinase-Associated Lipocalin and Cytokine Genes in Bone Marrow.

Background: Recently, various associations of NGAL with several hematological cancers have been reported. However, given that the regulation of NGAL gene expression by cytokines is tissue-specific, NGAL expression in relation to those of cytokine genes has not been analyzed in bone marrow (BM) tissue. The purpose of this study was to analyze the association between NGAL and 48 cytokine gene expression levels in mononuclear cells (MNCs) of BM at the time of diagnosis of hematological malignancy and to explore the expression pattern of NGAL and related cytokine genes in patients with hematological malignancies and controls. Methods: BM MNCs were isolated from 48 patients, who were classified as patients presenting myeloproliferative neoplasm, acute myeloid leukemia, myelodysplastic syndrome, and as controls. NGAL and cytokine genes were analyzed using NanoString. Data on hematological parameters were collected from medical records. Single and multiple regression analyses were performed to analyze relationships. Results: Normalized counts of 26 cytokine genes were related to NGAL normalized counts, while STAT3 and TLR4 normalized counts had the highest explanatory power. The following multiple regression model was developed: NGAL normalized counts=4316.825 + 9.056 × STAT3 normalized counts + 844.226 × IL5 normalized counts + 17.540 × TLR1 normalized counts - 28.206 × TLR2 normalized counts - 42.524 × IRAK4 normalized counts. In the multiple regression analysis, STAT3 and TLR4 normalized counts showed multicollinearity. NGAL, STAT3, IL5, and TLR4 normalized counts showed similar intergroup patterns. Conclusions: NGAL normalized counts was predicted by a multiple regression model, while they showed similar intergroup patterns to STAT3, IL5, and TLR4 normalized counts.

Analysis of neutrophil gelatinase-associated lipocalin, vascular endothelial growth factor, and soluble receptor for advanced glycation end-products in bone marrow supernatant in hematologic malignancies.

BACKGROUND: Inflammation is a known risk factor of cancer development, including inflammation-driven leukemogenesis. Evaluation of inflammation-related cytokines in early diagnosis stages is crucial to understand the development of hematologic malignancy. Our aim was to measure three cytokines- neutrophil gelatinase-associated lipocalin (NGAL), vascular endothelial growth factor (VEGF), and soluble receptor for advanced glycation end-products (sRAGE) in bone marrow (BM) samples from patients diagnosed with hematologic malignancy and compare these measurements with the control. Additionally, we evaluated whether NGAL was significantly associated with sRAGE, VEGF, and several hematological parameters. METHODS: BM samples were collected from 73 patients, who were classified into myeloproliferative neoplasm (MPN), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), plasma cell neoplasm (PCN) and control groups according to the BM smear and pathology review. An immunoassay, a Luminex assay, and an enzyme-linked immunosorbent assay were used to quantitate NGAL, VEGF, and sRAGE, respectively, while all measurements of NGAL, VEGF and sRAGE were performed on BM supernatants. Data on hematological parameters were collected from medical records. Intergroup comparisons were performed using the Kruskal-Wallis H-test and Pearson Chi-Square test. Single and multiple regression analyses were performed to analyze the relationships among the parameters. RESULTS: The independent factors associated with NGAL were neutrophil counts and VEGF. As for both NGAL and VEGF, the MPN (n = 23) group showed the highest level, while the MDS (n = 12) group showed low levels. NGAL levels in the AML (n = 13) and MDS groups were lower than in the control group (n = 14). The MPN group demonstrated higher VEGF levels than the AML and MDS groups. The MDS group showed lower VEGF levels than the PCN (n = 11) group. No statistical difference between the hematologic malignancy and control groups or among the hematologic malignancy groups was observed for sRAGE levels. CONCLUSION: NGAL was related to neutrophil count and VEGF. NGAL and VEGF showed similar intergroup patterns, reflecting that NGAL was associated with VEGF.

Analysis of bone marrow supernatant neutrophil gelatinase-associated lipocalin and hematological parameters in hematological malignancy.

BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is a urine biomarker related to acute renal injury. Whereas several studies have evaluated NGAL levels in hematological malignancy, using peripheral blood (PB). Recently, bone marrow (BM) NGAL level was reported to be higher than PB NGAL level in individuals with hematological malignancy, suggesting that BM NGAL would reflect BM microenvironment better than PB NGAL. We measured BM NGAL levels in patients with hematological malignancy, comparing those with NGAL levels in normal BM. We evaluated the association of BM NGAL with hematological parameters including neutrophil counts. METHODS: BM samples were collected from 107 patients who underwent BM examination. Immunoassays were used to assess NGAL levels. Data on hematological parameters were collected from medical records. Intergroup comparisons were performed using the Kruskal-Wallis H test and Pearson chi-square test. Single and multiple regression analyses were performed to analyze the relationships. RESULTS: The independent factors that affected the BM NGAL level were neutrophil counts and BM band neutrophil%, while neutrophil count was the main influencing factor. The acute myeloid leukemia (n = 18) and myelodysplastic syndrome (n = 25) groups showed statistically lower BM NGAL levels than patients with normal BM. The myeloproliferative neoplasm group (n = 34) showed higher BM NGAL levels than patients with normal BM, but this difference was not statistically significant. Neutrophil counts and BM band neutrophil% showed intergroup patterns similar to those of BM NGAL levels. CONCLUSION: BM NGAL was related to neutrophil count and BM band neutrophil%, showing different levels according to hematological malignant disease entities.

Association of peripheral blood neutrophil gelatinase-associated lipocalin levels with bone marrow neutrophil gelatinase-associated lipocalin levels and neutrophil count in hematologic malignancy.

BACKGROUND: Although neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker for acute kidney injury, recently, high NGAL levels have been reported in hematologic malignancies. Given the mechanism underlying NGAL synthesis and secretion in neutrophilic series, it is speculated that NGAL levels are higher in bone marrow (BM) than in peripheral blood (PB). Additionally, PB NGAL levels are thought to be associated with neutrophilic parameters. We aimed to test both hypotheses in hematologic malignancies. METHODS: Paired BM and PB samples were collected from 41 patients undergoing BM examination for hematologic malignancies. NGAL levels were measured using immunoassays. Data on hematologic parameters were collected from medical records. Single and multiple regression analyses were performed to analyze the relationship. RESULTS: PB and BM NGAL (n = 41) levels were significantly different (163.0 ± 258.3 and 413.1 ± 616.2 ng/mL [mean ± standard deviation], respectively; P < 0.05). Simple regression analysis and multicollinearity assessment showed that BM NGAL levels, BM neutrophil%, and neutrophil count were significant predictors of PB NGAL. Two multiple regression models were developed (model 1, PB NGAL = 21.467* neutrophil count - 0.785*BM neutrophil%; model 2, PB NGAL = 21.202*neutrophil count- 0.915*BM neutrophil% +0.10*BM NGAL). Akaike's information criterion and adjusted R2 values showed that model 1 had higher predictive accuracy for PB NGAL. In both models, neutrophil count was the only significant predictor. CONCLUSION: BM NGAL was significantly higher than PB NGAL in hematologic malignancy. In addition, PB NGAL could be expressed as a multiple regression model including neutrophil count and BM neutrophil%, being significantly influenced by neutrophil count.

Comparative evaluation of Plateletworks, Multiplate analyzer and Platelet function analyzer-200 in cardiology patients.

The objective of this study was to comparatively evaluate three commercial whole-blood platelet function analyzer systems: Platelet Function Analyzer-200 (PFA; Siemens Canada, Mississauga, Ontario, Canada), Multiplate analyzer (MP; Roche Diagnostics International Ltd., Rotkreuz, Switzerland), and Plateletworks Combo-25 kit (PLW; Helena Laboratories, Beaumont, TX, USA). Venipuncture was performed on 160 patients who visited a department of cardiology. Pairwise agreement among the three platelet function assays was assessed using Cohen's kappa coefficient and percent agreement within the reference limit. Kappa values with the same agonists were poor between PFA-collagen (COL; agonist)/adenosine diphosphate (ADP) and MP-ADP (-0.147), PFA-COL/ADP and PLW-ADP (0.089), MP-ADP and PLW-ADP (0.039), PFA-COL/ADP and MP-COL (-0.039), and between PFA-COL/ADP and PLW-COL (-0.067). Nonetheless, kappa values for the same assay principle with a different agonist were slightly higher between PFA-COL/ADP and PFA-COL/EPI (0.352), MP-ADP and MP-COL (0.235), and between PLW-ADP and PLW-COL (0.247). The range of percent agreement values was 38.7% to 73.8%. Therefore, various measurements of platelet function by more than one method were needed to obtain a reliable interpretation of platelet function considering low kappa coefficient and modest percent agreement rates among 3 different platelet function tests.

Report on the Project for Establishment of the Standardized Korean Laboratory Terminology Database, 2015.

The National Health Information Standards Committee was established in 2004 in Korea. The practical subcommittee for laboratory test terminology was placed in charge of standardizing laboratory medicine terminology in Korean. We aimed to establish a standardized Korean laboratory terminology database, Korea-Logical Observation Identifier Names and Codes (K-LOINC) based on former products sponsored by this committee. The primary product was revised based on the opinions of specialists. Next, we mapped the electronic data interchange (EDI) codes that were revised in 2014, to the corresponding K-LOINC. We established a database of synonyms, including the laboratory codes of three reference laboratories and four tertiary hospitals in Korea. Furthermore, we supplemented the clinical microbiology section of K-LOINC using an alternative mapping strategy. We investigated other systems that utilize laboratory codes in order to investigate the compatibility of K-LOINC with statistical standards for a number of tests. A total of 48,990 laboratory codes were adopted (21,539 new and 16,330 revised). All of the LOINC synonyms were translated into Korean, and 39,347 Korean synonyms were added. Moreover, 21,773 synonyms were added from reference laboratories and tertiary hospitals. Alternative strategies were established for mapping within the microbiology domain. When we applied these to a smaller hospital, the mapping rate was successfully increased. Finally, we confirmed K-LOINC compatibility with other statistical standards, including a newly proposed EDI code system. This project successfully established an up-to-date standardized Korean laboratory terminology database, as well as an updated EDI mapping to facilitate the introduction of standard terminology into institutions.

Comparison between Saliva and Nasopharyngeal Swab Specimens for Detection of Respiratory Viruses by Multiplex Reverse Transcription-PCR.

Nasopharyngeal swabs (NPSs) are being widely used as specimens for multiplex real-time reverse transcription (RT)-PCR for respiratory virus detection. However, it remains unclear whether NPS specimens are optimal for all viruses targeted by multiplex RT-PCR. In addition, the procedure to obtain NPS specimens causes coughing in most patients, which possibly increases the risk of nosocomial spread of viruses. In this study, paired NPS and saliva specimens were collected from 236 adult male patients with suspected acute respiratory illnesses. Specimens were tested for 16 respiratory viruses by multiplex real-time RT-PCR. Among the specimens collected from the 236 patients, at least 1 respiratory virus was detected in 183 NPS specimens (77.5%) and 180 saliva specimens (76.3%). The rates of detection of respiratory viruses were comparable for NPS and saliva specimens (P = 0.766). Nine virus species and 349 viruses were isolated, 256 from NPS specimens and 273 from saliva specimens (P = 0.1574). Adenovirus was detected more frequently in saliva samples (P < 0.0001), whereas influenza virus type A and human rhinovirus were detected more frequently in NPS specimens (P = 0.0001 and P = 0.0289, respectively). The possibility of false-positive adenovirus detection from saliva samples was excluded by direct sequencing. In conclusion, neither of the sampling methods was consistently more sensitive than the other. We suggest that these cost-effective methods for detecting respiratory viruses in mixed NPS-saliva specimens might be valuable for future studies.

Effect of Irradiation on Microparticles in Red Blood Cell Concentrates.

Changes in microparticles (MP) from red blood cell (RBC) concentrates in the context of irradiation have not been investigated. The aim of this study was to evaluate how irradiation affects the number of MPs within transfusion components. Twenty RBC concentrates, within 14 days after donation, were exposed to gamma rays (dose rate: 25 cGy) from a cesium-137 irradiator. Flow cytometry was used to determine the numbers of MPs derived from RBC concentrates before and 24 hr after irradiation. The mean number of MPs (±standard deviation) in RBC concentrates was 21.9×109/L (±22.7×109/L), and the total number of MPs ranged from 2.6×109/L to 96.9×109/L. The mean number of MPs increased to 22.6×109/L (±31.6×109/L) after irradiation. Before irradiation, the CD41-positive and CD235a-positive MPs constituted 9.5% (1.0×109/L) and 2.2% (263×106/L) of total MPs, respectively. After irradiation, CD41-positive MPs increased to 12.1% (1.5×109/L) (P=0.014), but the CD235a-positive MPs decreased to 2.0% (214×106/L) of the total MPs (P=0.369). Irradiation increases the number of CD41-positive MPs within RBC concentrates, suggesting the irradiation of RBC concentrates could be associated with thrombotic risk of circulating blood through the numerical change.

Evaluation of the Performance of a Fecal Tumor M2-PK Rapid Kit Using Stool Specimens for Detection of Colorectal Tumors.

BACKGROUND: In gastrointestinal tumors, only subunits of pyruvate kinase isoenzyme type M2 are detectable, and mainly in the dimeric form termed tumor M2-PK. A rapid test for the detection of M2-PK in stool has been developed. We evaluated the performance of the M2-PK rapid kit for the detection of colorectal tumors compared with colonoscopy DESIGN: Stool specimens (n=268) were obtained from patients who had a fecal occult blood test and colonoscopy performed for clinical evaluation or routine health checkup. Collected stool specimens were kept frozen at -20°C until testing. The fecal tumor M2-PK test was performed using a ScheBo M2-PK Quick test (ScheBo(®)•Biotech AG, Giessen, Germany) RESULTS: A total of 236 patients were analyzed, including 87 with tubular adenoma and 126 controls. In tumor M2-PK testing, 37 of 87 tubular adenoma specimens (42.5%) and 2 of 4 sessile serrated adenoma specimens (50%) tested positive. The specificity of the M2-PK rapid test was 87.3% CONCLUSIONS: The tumor M2-PK test demonstrated moderate sensitivity for detection of colorectal adenoma. M2-PK test could be considered as a useful point-of-care testing device for detection of colorectal tumors.

A Case Report of an Infant with Robertsonian Translocation (15;22)(q10;q10) and Literature Review.

Rob(15; 22) is rare and account for only 0.6% of all Robertsonian translocations. We describe a case with rob(15;22) in which the phenotype includes generalized hypotonia, respiratory distress, tent shaped upper lips, hyporeflexia and single umbilical artery. Chromosome analysis with peripheral blood was performed, while the karyotype was interpreted as 45,XX,der(15;22)(q10;q10). In Prader-Willi/Angelman Syndrome FISH studies, deletion of the SNRPN gene was not observed, but deletion of 15p11.2 was noted. Prader-Willi/Angelman Syndrome methylation-specific polymerase chain reaction and chromosomal microarrays showed negative findings. Molecular studies associated with spinal muscular atrophy and progressive muscular dystrophy also showed negative findings. We suggest that rob(15;22) and deletion of 15p11.2 could be related to clinical presentation like this case.

Measurement of platelet aggregation functions using whole blood migration ratio in a microfluidic chip.

Platelets play a major role in maintaining endothelial integrity and hemostasis. Of the various soluble agonists, ADP is an important in vivo stimulus for inducing platelet aggregation. In this study, a simple, rapid, and affordable method was designed for testing bleeding time (BT) and platelet aggregation with a two-channel microfluidic chip. Whole blood migration ratio (MR) from a microchip system was evaluated in comparison to the closure time (CT) from PFA-100 assays (Siemens, Germany) and CD62P expression on platelets. To induce platelet aggregation, a combination of collagen (1.84 mg/ml) and ADP (37.5 mg/ml) were used as agonists. After adding the agonists to samples, whole blood MR from the microchip system was measured. The outcome of the assessment depended on reaction time and agonist concentration. MR of whole blood from the microchip system was significantly correlated with CT from PFA-100 (r = 0.61, p <  0.05, n = 60). In addition, MR was negatively correlated with CD62P expression (r =-0.95, p <  0.05, n = 60). These results suggest that the measurement of MR using agonists is an easy, simple and efficient method for monitoring platelet aggregation in normal and ADP-receptors defective samples, along with the BT test. Thus, usage of the current microfluidic method could expand to diverse applications, including efficacy assessments in platelet therapy.

A Case of Diabetic Mellitus Foot Infection by a Newly Reported Neisseria Skkuensis: Case Report.

In pus and wound samples collected from the right second toe of a 61-year-old woman with diabetes mellitus (DM), gram-negative diplococci bacterium was observed. However, the bacterium could not be identified by conventional microbiological methods and mass spectrometry. In the partial 16S rRNA gene sequence analysis, the bacterium showed a 100% identity match with GenBank sequence FJ0763637.1 (Neisseria skkuensis). N. skkuensis, SMC-A9199 strain, was reported as a novel species in 2010 based on its phenotypic characteristics and the 16S rRNA gene sequence, which was isolated from the blood and wound pus of a DM patient with a foot ulcer. The second reported N. skkuensis was identified from the blood cultures of a patient with endocarditis. To the best of our knowledge, this is only the third report of N. skkuensis.

Effects of dietary Forsythia suspensa on feed utilization, rumen fermentation, and immune response of Korean native goats (Capra hircus) / Efectos de la adición dietaria de Forsythia suspensa sobre utilización del alimento, fermentación ruminal y respuesta inmune en cabras nativas coreanas (Capra hircus) / Efeitos na dieta pela utilização de Forsythia suspensa nos alimentos, fermentação ruminal e resposta imune em caprinos nativos coreanos (Capra hircus)

Background: plant extracts of Forsythia suspensa, which contain pinoresinol, have been proposed as an alternative to antibiotics due to their antioxidant, anti-inflammatory, and microflora modulating effects. Objective: to evaluate the effects of dietary F. suspensa on feed utilization, rumen fermentation, blood parameters and immune response of Korean native goats (Capra hircus). Methods: a total of nine Korean native goats were assigned to one of three dietary treatments: 1) a basal diet without F. suspensa, 2) a basal diet added with 0.25% F. suspensa, or 3) a basal diet added with 0.50% F. suspensa. A change-over design was used in three consecutive periods to give 9 replicates per treatment. Each period lasted 18 days, including 15 days of adaptation to feed and environmental conditions and three days of total collection of feces and urine. Rumen fluid and blood samples were also collected. Nutrient digestibility, nitrogen retention, ruminal content, and blood metabolites—including immunoglobulins—were measured. Results: F. suspensa supplementation had no effect on nutrient digestibility, whole body nitrogen retention rate, ruminal pH, acetate, propionate, isobutyrate, butyrate, isovalerate, or valerate content (p>0.05). However, F. suspensa supplementation decreased total volatile fatty acids (VFA) concentration compared with the control diet, regardless of F. suspensa concentrations (p<0.05). Goats fed a diet containing 0.25% F. suspensa had greater alkaline phosphatase (ALP) concentration than those fed a control or a diet with 0.50% F. suspensa (p<0.05). Feeding different concentrations of dietary F. suspensa did not influence plasma IgA and IgM levels (p>0.05), although goats receiving both 0.25% and 0.50% F. suspensa diets had greater plasma IgG than those fed the control diet (p<0.05). Conclusion: supplementation of 0.25% F. suspensa significantly decreased total VFA concentrations and increased plasma IgG in Korean native goats, compared with those fed the control diet.

Antecedentes: los extractos de la planta Forsythia suspensa contienen pinoresinol. Dicha planta se ha propuesto como una alternativa a los antibióticos debido a su contenido de antioxidantes, antiinflamatorios y sus efectos moduladores de la microflora. Objetivo: evaluar los efectos dietarios de F. suspensa sobre la utilización del alimento, fermentación ruminal, parámetros sanguíneos y respuesta inmune en cabras nativas coreanas (Capra hircus). Métodos: un total de nueve cabras nativas coreanas fueron asignadas a 1 de 3 tratamientos: (1) una dieta basal sin F. suspensa, (2) una dieta basal con 0,25%, o (3) 0,50% de F. suspensa dietaría en un diseño change-over por tres periodos consecutivos con nueve réplicas por tratamiento. Cada periodo se prolongó por 18 días, incluyendo 15 días de adaptación al alimento y a las condiciones ambientales, y tres días de colección total de heces y orina. Tambien se colectó fluido ruminal y muestras sanguineas. Se midió la digestibilidad de nutrientes, nitrógeno retenido, contenido ruminal y metabolitos sanguíneos —incluyendo inmunoglobulinas—. Resultados: la suplementación con F. suspensa no tuvo efecto sobre la digestibilidad de los nutrientes, la tasa de retención de N, pH ruminal, ni sobre los contenidos de acetato, propionato, isobutirato, butirato, isovalerato o valerato (p>0,05). Sin embargo, la suplementación con F. suspensa disminuyo la concentración total de ácidos grasos volátiles (VFA) en comparacion con la dieta control, sin tener en cuenta las concentraciones de F. suspensa (p<0,05). Las cabras alimentadas con la dieta de 0,25% de F. suspensa tuvieron mayor concentración de fosfatasa alcalina (ALP) que cuando fueron alimentadas con la dieta control o con la de 0,50% de F. suspensa (p<0,05). Aunque la alimentación con diferentes concentraciones de F. suspensa no influyo en los niveles de IgA y IgM en plasma (p>0,05), las cabras que recibieron 0,25 y 0,50% de F. suspensa tuvieron mayor concentración de IgG en plasma que aquellas alimentadas con la dieta control (p<0,05). Conclusiones: la suplementación de 0,25% de F. suspensa disminuyo significativamente la concentración total de VFA e incrementó la IgG en el plasma de las cabras nativas coreanas, en comparación con aquellas que consumieron la dieta control.

Antecedentes: a Forsythiae suspensa é uma planta e seu extrato contém pinoresinol. Tem sido proposta como uma alternativa aos antibióticos existentes, devido ao seu teor de antioxidantes, anti-inflamatórias e efeitos moduladores da microflora, com base em estudos feitos in vivo e in vitro. Objetivo: avaliar os efeitos na dieta da inclusão de F. suspensa no aproveitamento alimentar, fermentação ruminal, parâmetros sanguíneos e resposta imune em caprinos coreanos nativos (Capra hircus). Métodos: um total de nove cabras coreanas nativas foram assignadas a um de três tratamentos na dieta: (1) uma dieta basal sem F. suspensa, (2) uma dieta basal com 0,25% de F. suspensa, ou (3) com 0,50% de F. suspensa na dieta. Se fez um desenho change-over por três períodos consecutivos com nove repetições por tratamento. Cada período de pesquisa consistiu de 18 dias, incluindo 15 dias de adaptação às condições alimentares e ambientais, e três dias de coleta total de fezes e urina. Também foram coletadas amostras de líquido ruminal e sangue. Mediou-se a digestibilidade dos nutrientes, o nitrogênio (N) retido, o conteúdo ruminal, e os parâmetros sanguíneos —incluindo imunoglobulinas—. Resultados: a suplementação com F. suspensa não teve nenhum efeito sobre a digestibilidade dos nutrientes, a taxa de retenção de N, o pH do rúmen, nem sobre os conteúdos de: acetato, propionato, isobutirato, butirato, isovalerato e valerato (p>0,05). No entanto, a suplementação de F. suspensa diminuiu a concentração total de ácidos gordurosos voláteis (AGV) em comparação com a dieta controle, independentemente das concentrações de F. suspensa (p<0,05). As cabras alimentadas com 0,25% F. suspensa na dieta tiveram maiores quantidades da enzima fosfatasse alcalina (ALP) do que quando elas foram alimentadas com a dieta controle ou com 0,50% de F. suspensa (p<0,05). Ainda que a alimentação com diferentes níveis de F. suspensa não influenciou os níveis de IgA e IgM no plasma (p>0,05), as cabras que receberam na dieta 0,25 e 0,50% de F. suspensa tiveram uma maior concentração de IgG no plasma comparadas as cabras alimentadas com a dieta controle (p<0,05). Conclusões: a suplementação com 0,25% e 50% diminuiu significativamente a concentração de AGV e incrementou a concentração em plasma de IgG em cabras nativas coreanas comparadas com aquelas que consumiram a dieta controle.

Effect of Interleukin-29 on Interferon-α Secretion by Peripheral Blood Mononuclear Cells.

OBJECTIVE: The effect of interleukin (IL)-29, a new therapeutic agent similar to type I interferons (IFNs), on IFN-α secretion of human plasmacytoid dendritic cells (pDCs) has not been studied. Therefore, in this study, we aimed to clarify the effect of IL-29 on IFN-α secretion of pDCs using human peripheral blood mononuclear cells (PBMCs) in the presence of cytosine-phosphate-guanosinemotif-containing oligodeoxy nucleotides (CpG). MATERIALS AND METHODS: In this experimental and prospective study, PBMCs were ob- tained from 11 healthy volunteers and divided into four culture conditions: I. control, II. CpG treatment, III. IL-29 treatment and IV. CpG plus IL-29 treatment. The amount of IFN-α secretion was measured from each culture supernatant by flow cytometry using the flowcytomix apparatus (eBioscience, Vienna, Austria). Fractional IFN-α production of the cultured PBMCs was measured by intracellular staining using the cytomics FC 500 system (Beckman Coulter, Brea, CA, USA) with CXP Software. RESULTS: The mean ± standard deviation (SD) of supernatant IFN-α secretion per pDC/µL was 5.7 ± 9.3 pg/mL/count/µL for condition I, 1071.5 ± 1026.6 pg/mL/count/µL for condition II, 14.1 ± 21.1 pg/mL/count/µL for condition III, and 1913.9 ± 1525.9 pg/mL/count/µL for condition IV. There were statistically significant differences between conditions I and II as well as betweenconditions II and IV. Intracellular IFN-α production was only detectable in the pDC fraction from one culture; the production amount was similar between the cells treated with CpG and those treated with CpG plus IL-29. Natural killer (NK) cell production of IFN-α was observed in two out of three cultures and one culture showed IFN- α production in the monocyte fraction. CONCLUSION: IL-29 alone did not show any effect on IFN-α secretion of PBMCs. However, the addition of CpG along with IL-29 enhanced IFN-α secretion of PBMCs. Given that pDCs are the major secretors of IFN-α in peripheral blood, this result has suggested the possibility that IL-29 has an enhancing effect in human pDC IFN-α secretion. Although the supernatant IFN-α secretion was not directly correlated with pDCs's intracellular IFN-α production in this study, prolonged incubation of pDC and other PB subsets with CpG or IL-29 for over 4 hours could be applied in future studies. These studies would help to elucidate the mechanism of action of IL-29 in human pDCs associated with viral infections.

A case report of a fetus with mosaic autosomal variegated aneuploidies and literature review.

Mosaic variegated aneuploidy (MVA) is a recessive condition characterized by mosaic aneuploidies, predominantly trisomies and monosomies, involving multiple chromosomes and tissues. The phenotype of MVA syndrome includes severe microcephaly and growth deficiency, central nervous system anomalies, mental retardation, mild physical anomalies, and predisposition to cancer. We report a case of true fetal mosaicism for variegated aneuploidies detected in amniotic fluid cells. A 33-year-old primigravida woman at 5 weeks 1 day of gestation was referred to our tertiary hospital because of a high-risk pregnancy associated with IgA nephropathy. In a quadruple screening test performed at the 15(th) week of gestation, alpha fetoprotein was 73.4 IU/mL (2.792 MoM), suggesting that she was at high risk of neural tube defect. Following amniocentesis performed at the 17 weeks' gestation, chromosome examination of amniocyte culture showed premature chromatic separation in 63% of the metaphases (58/92) and a high frequency of gain and loss of chromosomes. Repeat amniocentesis at 21 weeks' gestation consistently showed the presence of multiple mosaic autosomal variegated aneuploidies. Ultrasonography at 21 weeks' gestation revealed relatively small head circumference for gestational age (<3%) and vermis defect, suggesting that the fetus would have microcephaly and Dandy-Walker malformation. Cytogenetic analysis with peripheral blood of the parents showed normal karyotype. In summary, we hereby report the cytogenetic analysis and prenatal findings of MVA.

Clinical performance evaluation of the Sofia RSV FIA rapid antigen test for diagnosis of respiratory syncytial virus infection.

A recently introduced Sofia respiratory syncytial virus (RSV) fluorescent immunoassay (FIA) was evaluated against the BinaxNOW RSV card and the SD Bioline RSV test using 348 respiratory samples. The Sofia, BinaxNOW, and SD Bioline kits showed sensitivities of 66%, 65%, and 64%, respectively, for detecting RSV-A, and 71%, 63%, and 65% for detecting RSV-B, respectively.

Flow cytometric enumeration of parasitemia in cultures of Plasmodium falciparum stained with SYBR Green I and CD235A.

A flow cytometric (FACS) detection method for Plasmodium falciparum cultures (P. falciparum) was developed using SYBR Green I and CD235A and compared against the Giemsa stained microscopic examination. The cultured P. falciparum were spiked into red blood cells (RBCs) to yield parasitemia, ranging from 0.01% to 22.0%. FACS analysis demonstrated a clear separation between P. falciparum infected and uninfected RBCs. The measured percentage of parasitemia by FACS revealed higher precision (CV of 2.2-37.2%) with the sensitivity of 0.01% parasitemia than Giemsa stained microscopic examination (CV of 7.2-66.0%). High correlation of measured parasitaemia (r=0.98, P<0.05) was observed between FACS and Giemsa stained microscopic analyses. The higher levels of parasitaemia detection were observed in all ranges by FACS in comparison to Giemsa stained microscopic analysis. The currently reported FACS method using SYBR Green I and CD235A is potentially useful for measuring parasitemia in treating patients.